Chromatography

JOURNAL OF CHROMATOGRAPHY LIBRARY- volume 56
chromatography in the
petroleum industry
This Page Intentionally Left Blank
JOURNAL OF CHROMATOGRAPHY LIBRARY-volume 56
chromatography in the
petroleum industry
edited b y
E.R. Adlard
Burton, South Wirral, UK
ELSEVIER
Amsterdam-Lausanne-New York-Oxford-Shannon -Tokyo 1995
ELSEVIER SCIENCE B.V.
Sara Burgerhartstraat 25
P.O. Box 21 1,1000AE Amsterdam, The Netherlands
Library of Congress Cataloging-in-Publication Data
Chromatography in the petroleum industry / edited b y E.R. Adlard.

p. cm. -- (Journal of chromatography library ; v. 56)
Includes bibliographical references and index.
ISBN 0-444-89776-3(acid-free)
1. Petroleum-Analysis. 2. Chromatographic analysis-Industrial
applications. I. Adlard, E.R. 11. Series.
TP691.C58 1995
665.50287--dc?O 95-30628
CIP
ISBN 0-444-89776-3
01995 Elsevier Science B.V. All rights reserved.
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V
Contents
Foreword .................................................................................................................... XV
List of Contributors..................................................................................................... XVII
Chapter 1 . The analysis of hydrocarbon gases .................................................. 1

C.J. Cowper
1.1 Introduction ....................................................................................................... 1
1.2 Natural gas ......................................................................................................... 2
1.2.1 Analytical requirements ...................................................................... 5
1.2.1.1 CV measurement ................................................................ 5
1.2.1.2 Hydrocarbon dewpoint calculation..................................... 8
1.2.2 Analytical procedures.......................................................................... 10
1.2.2.1 Isothermal methods............................................................. 10
1.2.2.2 Two detectors ..................................................................... 16
1.2.2.3 C6+ detail............................................................................ 16
1.2.2.4 Temperature programming................................................. 18
1.2.2.5 Combined systems.............................................................. 19
1.2.2.6 Separation in backflush ...................................................... 19
1.2.2.7 Rapid analysis..................................................................... 21
1.2.3 Quantitative measurement................................................................... 25
1.2.3.1 Comprehensive analysis ..................................................... 25
1.2.3.2 Air contamination............................................................... 25
1.2.3.3 Resolution........................................................................... 26
1.2.3.4 Precision ............................................................................. 26
1.2.4.5 Response function .............................................................. 27
1.3 Refinery gas ....................................................................................................... 28
1.3.1 Analytical requirements ...................................................................... 29
1.3.2 Analytical procedures.......................................................................... 30
1.3.3 Sample handling .................................................................................. 37
1.4 Conclusions ....................................................................................................... 38
1.5 Acknowledgements............................................................................................ 39
1.6 References ......................................................................................................... 40
Chapter 2 .Advances in simulated distillation ................................................... 41

D.J. Abbott
2.1 Introduction....................................................................................................... 41
2.2 Middle distillates and lube oils .......................................................................... 42
2.2.1 Precision.............................................................................................. 42
2.2.2 Capillary columns ............................................................................... 44
VI Contents
2.2.3 Aromatics and heteroatoms ................................................................. 44

2.2.4 Multi-element speciation.,................................................................... 45
2.3 Gasolines and gasoline fractions ....................................................................... 46
2.4 Heavy lube oils and residues ..... ................................................................. 47
2.5 Crude oils ................... .................................................................................. 50
2.6 Process control and other applications .............................................................. 51
2.1. Conclusions ........... ....................................................................... 52
2.8 References ....................................... .................................................... 52
Chapter 3 . The chromatographic analysis of refined and synthetic waxes ..... 55

A . Barker
3.1 Introduction .................................................... ............................... 55
3.2 Gas liquid chromatography ......... ................................................................. 57
3.2.1 Establishing present technology .......................................................... 57
3.2.2 The 1980s revolution .......................................................................... 59
3.2.2.1 Sample introduction ............................................................ 60
3.2.2.2 Detection ............................................................................ 64
3.2.2.3 High temperature GLC columns ......................................... 65
3.2.3 Quantitative gas liquid chromatography separation of waxes ............. 70
3.2.3.1 Carbon number distribution analysis .................................. 75
3.3 Supercritical fluid chromatography of waxes .................................................... 78
3.4 Size exclusion chromatography ......................................................................... 81
3.4. I Early work on the SEC analysis of waxes ........................................... 82
3.4.2 Present day technology ........................................................................ 84
3.5 Conclusions ............................... .................................... 89
3.6 References ............. .................................................... 90
Chapter 4 .Hydrodynamic chromatography of polymers ................................. 95

.
J Bos and R Tijssen .
4.1 Introduction ............................... ......................................................... 95
4.2 Microcapillary hydrodynamic chromatography ................................................. 96
4.2.1 The0ry ................................................................................................. 96
4.2.2 Expressing the size of macromolecules ............................................... 99
4.2.3 Instrumentation ................................................................................... 100
4.2.3.1 General aspects ................. ..................................... 100
4.2.3.2 Detection ........ .................................................... 102
4.2.3.3 The column ......................................................................... 103
4.2.4 Applications ........................................................................................ 104
4.3 Capillary hydrodynamic chromatography.......................................................... 111
4.3.1 From hydrodynamic chromatography to tubular pinch ....................... 111
4.3.2 Coiling effects ........... ...................................................... 117
4.3.3 Applications ........................................................................................ 119
4.4 Hydrodynamic chromatography in packed columns .......................................... 120
4.5 Conclusions ............................................................................................ 124
4.6 References ................................. ........................................... 125
Contents VII
Chapter 5 . Chromatography in petroleum geochemistry ................................. 127

S.J. Rowland and A.T. Revill
5.1 Introduction ....................................................................................................... 127
5.1.1 Recent reviews .................................................................................... 127
5.2 Kerogen and other petroleum macromolecules ................................................. 128
5.3 Geochemistry of petroleum ............................................................................... 130
5.3.1 LC, TLC and TLC-FID ....................................................................... 131
5.3.2 HPLC .................................................................................................. 132
5.3.3 GC ....................................................................................................... 133
5.3.4 GC-MS ................................................................................................ 135
5.3.5 GC-isotope ratio-MS ........................................................................... 137
5.3.6 LC-MS ................................................................................................ 137
5.3.7 SFC, GPC and SEC............................................................................. 138
5.4 Summary............................................................................................................ 138
5.5 References ......................................................................................................... 139
Chapter 6 .The O-FID and its applications in petroleum product analysis .... 143
A.Sironi and G.R. Verga
6.1 Introduction ....................................................................................................... 143
6.2 Oxygenates as components of motor gasoline ................................................... 144
6.2.1 Determination of oxygenates in unleaded fuels................................... 146
6.3 O-FID analyser .................................................................................................. 147
6.3.1 Cracking reactor .................................................................................. 147
6.3.1.1 Low temperature cracker .................................................... 150
6.3.2 Hydrogenation microreactor ............................................................... 151
6.4 Analytical procedure ......................................................................................... 152
6.4.1 Quantitative analysis ........................................................................... 152
6.4.2 Total oxygen determination................................................................. 154
6.4.2.1 Selectivity for oxygenates and sensitivity........................... 155
6.5 O-FID applications............................................................................................ 156
6.6 Conclusion......................................................................................................... 157
6.7 References ......................................................................................................... 157
Chapter 7 . Microwave plasma detectors ............................................................ 159

.
A de Wit and J Beens .
7.1 Introduction....................................................................................................... 159
7.2 Principle of operation of an atomic emission detector (AED) ........................... 160
7.3 Historical development of the plasma detector.................................................. 162
7.4 Description and evaluation of a home-built atomic emission detector .............. 164
7.4.1 Description of the apparatus................................................................ 164
7.4.1.1 Microwave cavities............................................................. 164
7.4.1.2 Microwave power supply ................................................... 166
7.4.1.3 Spectrometer....................................................................... 167
7.4.1.4 Optical system .................................................................... 168
7.4.1.5 Sample introduction system................................................ 169
VllI Contents
Plasma start-up ....................................................................................

7.4.2 169
Operating limits of the detector ...........................................................
7.4.3 170
7.4.3.1 Emission line intensity ........................................................ 171
7.4.3.2 Plasma pressure .......... ......................... 172
7.4.3.3 Microwave power ............................................................... 172
7.4.3.4 Quartz tube diameter .......................................................... 173
7.4.3.5 Optical system .................................................................... 173
7.4.3.6 Slit width .............................. .................................... 174
7.4.3.7 Upper limit of detection...................................................... 174
7.4.3.8 Type and amount of scavenger gas ..................................... 176
7.4.3.9 Linear dynamic range of the detector ................................. 177
7.4.3.10 Linear dynamic range for hydrogen.................................... 179
7.4.4 Selectivity............................................................................................ 180
7.4.4.1 Selectivity to H, 0 and N .................................................... 180
7.4.4.2 Selectivity to carbon ........................................................... 181
7.5 Conclusions ....................................................................................................... 184
7.6 Description of the Hewlett Packard 592 1A AED .............................................. 184
7.6.1 Gas chromatograph and transfer line ................................................... 185
7.6.2 Microwave cavity, discharge tube and gas flow system ...................... 187
7.6.3 The photodiode array spectrometer ..................................................... 189
7.6.4 Computerized control and data treatment............................................ 190
7.6.5 Characteristics ..................................................................................... 190
7.7 Typical applications .................. ............................................................... 191
7.7.1 Multi-element SimDist ses .......................................................... 191
7.7.1.1 Multi-element SimDist s o h a r e ......................................... 192
7.7.1.2 Linear dynamic ranges ........................................................ 194
7.7.1.3 Simdist results .................................................................... 195
7.7.2 Determination of chlorine-containing compounds in the ethylene
oxide process ....................................................................................... 197
7.7.3 Determination of oxygenates in gasoline ............................................ 197
7.7.4 Determination of noble gases in natural gas ........................................ 197
7.7.5 Determination of metal porphyrins in crude oils ................................. 197
7.5 Conclusion................. .................................................... 197
7.9 References ........................................................................... 200
Chapter 8. The sulfur chemiluminescence detector........................................... 201

R.S. Hutte
8.1 Introduction ....................................................................................................... 201
8.2 Sulfur-selective detectors for gas chromatography............................................ 202
8.3 The sulfur chemiluminescence detector ............................................................. 203
8.4 Performance characteristics of the SCD ............................................................ 207
8.5 Factors influencing the sensitivity and selectivity of the SCD ........................... 209
8.6 Flameless sulfur chemiluminescence................................................................. 211
8.7 Column selection and sampling techniques ....................................................... 213
8.8 ADdications ...................................................................................................... 215
Contents IX
Refinery gases .....................................................................................

8.8.1 215
8.8.1.2 Gasoline.............................................................................. 218
8.8.1.3 Diesel fuels ......................................................................... 218
8.8.1.4 High temperature gas chromatography............................... 218
8.9 Conclusions ....................................................................................................... 227
8.10 Acknowledgments ............................................................................................. 227
8.11 References ......................................................................................................... 227
Chapter 9. Multi-column systems in gas chromatography ............................... 231

.
H.Mahler. T Maurer and F.Mueller
9.1 Introduction ....................................................................................................... 231
9.1.1 Limitations of single-column systems ................................................. 231
9.1.2 Multi-column systems ......................................................................... 232
9.1.3 Multi-column chromatography............................................................ 233
9.1.3.1 Definition............................................................................ 233
9.1.3.2 Variants of column switching ............................................. 234
9.2 Selectivity tuning in series-coupled columns..................................................... 238
9.3 Column switching techniques ............................................................................ 241
9.3.1 General................................................................................................ 241
9.3.2 Backflushing ....................................................................................... 244
9.3.3 cutting ................................................................................................. 245
9.3.4 Distribution cutting ............................................................................. 246
9.3.5 Special switching techniques............................................................... 249
9.3.5.1 Summation of backflushed compounds .............................. 249
9.3.5.2 Stopped flowlstuttering....................................................... 250
9.3.5.3 Recycle chromatography .................................................... 250
9.4 Practical aspects ................................................................................................ 250
9.4.1 Valve switching................................................................................... 251
9.4.2 Valveless flow switching..................................................................... 252
9.4.2.1 Deans-switching ................................................................. 253
9.4.2.2 Live-switching.................................................................... 255
9.4.3 Column-switching as a sampling technique......................................... 258
9.4.3.1 Sampling fiom a capillary pre-column to a main capillary
column ................................................................................ 260
9.4.3.2 Coupling of columns of different type with intermediate
trapping............................................................................... 262
9.5 Strategies for the application of multi-column systems ..................................... 263
9.5.1 General remarks .................................................................................. 263
9.5.2 Guidelines for the use of single- or multi-column systems.................. 266
9.6 References ......................................................................................................... 266
Chapter 10 .
Supercritical fluid extraction ........................................................... 269
T.P. Lynch
10.1 Introduction ....................................................................................................... 269
10.2 Why use supercritical fluid extraction? ............................................................. 271
x Contents
10.2.1 Technical advantages .. .................................................. 27 1

10.2.1. 1 Solubility ........................ ........................................... 271
10.2.1.2 Selectivity ........................................................................... 271
10.2.1.3 Volatility ............................................................................. 274
10.2.1.4 Matrix penetration .............................................................. 274
10.2.2 Environmental and safety advantages ................................................. 275
10.3 ................................................................ 275
ms ................................................................................. 276
10.3.2 Sample vessels and temperature control .............................................. 277
10.3.3 Depressurization .............................. 278
10.3.4 Extract collection ................................................................................ 278
10.4 SFE techniques ............... ............................................... 280
10.4.1 Static extraction . .................................................... 280
10.42 Dynamic extract ............................................... 280
10.4.3 Recirculating extraction ...................................................................... 281
10.4.4 Extraction of liquids .. ........................................ 281
10.4.5 Reactive extraction .... ........................................ 283
10.4.6 Off-line extraction ............................................................................... 283
10.4.7 On-line extraction...... ........................................ 284
10.5 Petroleum-based applications ...... ............................................................. 284
10.5.1 Off-line applications ............................................................................ 284
10.5.1.1 Residual oil on drill cuttings ............................................... 284
10.5.1.2 Drilling mud characterization ............................................. 286
10.5.1.3 Petroleum core and rock extractions................................... 287
10.5.1.4 Refinery catalysts, deposits and sludges ............................. 288
10.5.1.5 Automotive engine particulates .......................................... 288
10.5.1.6 Environmental analysis ....................................................... 290
10.5.2 On-line applications ............................................................................ 291
10.5.2.1 On-line SFE-GC ................................................................. 291
10.5.2.2 On-line SFE-HPLC............................................................. 299
10.6 Conclusion ......................................................................................................... 301
10.7 References .. .............. ............................... 301
Chapter 11. Supercritical fluid chromatography ................................................ 305

.
1 Roberts
1 1 . 1 Introduction ..... ................................................................... 305
1 1.2 Instrumentation .................................................................................................. 306
1 I .2.1 Mobile phase pumps ........................................................................... 307
1 1.2.I . 1 Reciprocating pumps ........ ...................................... 308
1 1.2.1.2 Syringe pumps .................................................................... 308
I 1.2.2 Ovendtemperature control ................ ...................................... 309
11.2.3 Injectors .............................................................................................. 309
1 1.2.4 Detectors . ................................................................ 310
11.2.4.1 Universal detectors ............................................................. 311
1 1.2.4.2 Spectroscopic detectors ...................................................... 314
Contents XI
11.2.4.3 Element specific detection.................................................. 318

11.2.5 Restrictors ........................................................................................... 320
11.2.5.1 Fixed restrictors.................................................................. 320
11.2.5.2 Variable restrictors ............................................................. 321
1 1.3 Applications ...................................................................................................... 322
11.3.1 Simulated distillation........................................................................... 322
11.3.2 Hydrocarbon group type analysis........................................................ 328
11.3.2.1 Gasolines ............................................................................ 330
11.3.2.2 Kerosenes and naphthas...................................................... 334
11.3.2.3 Diesel fuels ......................................................................... 336
11.4 References ......................................................................................................... 343
Chapter 12 .
HPLC and column liquid chromatography .................................... 347
A.C. Neal
12.1 Introduction....................................................................................................... 347
12.2 Apparatus .......................................................................................................... 348
12.2.1 Solvent reservoirs ................................................................................ 348
12.2.2 Pumps.................................................................................................. 348
12.2.3 Sample injectors.................................................................................. 350
12.2.4 Columns .............................................................................................. 351
12.2.5 Detectors ............................................................................................. 353
12.2.6 Selective property detectors ................................................................ 353
12.2.6.1 UV-visible spectrophotometers .......................................... 353
12.2.6.2 Diode array detectors (DAD) ............................................. 354
12.2.6.3 Fluorescence detectors........................................................ 355
12.2.6.4 Electrochemical detectors................................................... 355
12.2.6.5 Flame ionization detector ................................................... 356
12.2.6.6 Mass spectrometers ............................................................ 356
12.2.6.7 Infrared and NMR .............................................................. 357
12.2.7 Bulk property detectors....................................................................... 358
12.2.7.1 Refractive index detector.................................................... 358
12.2.7.2 Evaporative light scattering detectors................................. 359
12.2.7.3 Dielectric constant detector ................................................ 360
12.3 Quantitation....................................................................................................... 361
12.4 Applications ...................................................................................................... 362
12.4.1 Individual compounds ......................................................................... 362
12.4.1.1 Polycyclic aromatic hydrocarbons (PAHs)......................... 362
12.4.1.2 Other indigenous compounds ............................................. 365
12.4.1.3 Additives and contaminants................................................ 366
12.4.1.4 Compound classes .............................................................. 368
12.5 Preparative HPLC and column liquid chromatography..................................... 370
12.5.1 Standard methods ................................................................................ 370
12.6 Individual publications ...................................................................................... 371
12.7 Future trends...................................................................................................... 372
12.8 References ......................................................................................................... 372
XI1 Contents
Chapter 13. Modern data handling methods ....................................................... 375

N. Dyson
1 3.1 Introduction ............................... ...................................... 375
13.2 The role ofthe data processor ........................................................................... 376
13.2.1 The modem integrator has four distinct roles ..... .......................... 376
13.2.2 Lab economics .................................................................................... 378
13.2.3 Prepare and distribute data, information and reports ........................... 378
13.3 Limitations of the detector signal ......... .................................................. 378
13.3.1 Injector fractionation is uniform .................... ............................. 378
13.3.2 All solutes of inte ........................ 379
13.3.3 Column resolution is ........................ 379
13.3.4 All solutes of interest ........................ 379
13.3.5 Signalinoise ratio ........................ 379
13.3.6 Detector is linear ............................. 380
13.4 Detector signal processing .... ........................ 380
13.4.1 Improving signal to ........................ 380
13.4.2 Electronic filtering ............................ .......................... 381
13.4.3 Software smoothing ..... .............................................
13.4.4 Measurement of pe ........................ 381
13.4.5 Baseline placement ....... ......................................
I 3.4.6 Mathematical separation (deconvolution) of peaks and integrator
developments ................................... ...................................... 382
13.4.7 Selective extraction ..... ................................................. 383
13.4.7.1 Peak model1 ...................................... 383
13.4.7.2 Signal subtraction ............................................................... 383
13.4.8 Resolution enhancement. .... .................................................... 384
13.5 Measurement errors .................... 384
13.5.1 Accuracy
13.5.2 Errors crea .......................... 385
13.5.3 Errors crea
13.5.4 Errors created by asymmetry ...................... .................... 386
13.5.5 Transmission of systematic errors through oups ............. 387
13.5.6 Baseline construction errors ................................................................ 390
13.5.7 Errors of tangent/perpendicular peak splitting .................................... 390
33.5.8 Ease of use ............................... ................................................ 390
13.6 Calibration ......... ................................................ 390
13.6.1 Use of area or height for quantitation .................................................. 390
13.6.2 Calculations and response factors ........................................
13.6.3 Limitations of the standard calculations .............................................. 391
................................................. 391
................................................. 392
13.6.3.3 External standard ................................................
13.6.4 Calibration curves ............................. .......................................... 392
13.6.5 Empirical correction of analysis errors ................................................ 393
13.7 Validation and standard chromatograms ........................................................... 393
Contents XI11
13.7.1 Meaning of validation ......................................................................... 393

13.7.2 System suitability................................................................................ 394
13.7.3 Validation and standard chromatograms............................................. 394
13.8 Strategies for peak measurement ....................................................................... 395
13.8.1 Noise ................................................................................................... 396
13.8.2 Baseline drift ....................................................................................... 396
13.8.3 Peak overlap........................................................................................ 396
13.8.4 Asymmetry .......................................................................................... 396
13.8.5 Detectors ............................................................................................. 397
133.6 Integrators ........................................................................................... 397
13.8.7 Checking the analysis results ............................................................... 397
13.8.7.1 Checking the chromatogram............................................... 397
133.7.2 Checking the analysis rep0rt ............................................... 398
13X7.3 Accepting the results .......................................................... 398
13.9 References ......................................................................................................... 398
Chapter 14 .
Capillary electrophoresis in the petroleum industry ...................... 401
.
T Jones and G Bondoux .
14.1 Introduction ....................................................................................................... 401
14.2 Separation techniques ........................................................................................ 404
14.2.1 Free-zone capillary electrophoresis(FZCE) ....................................... 404
14.2.2 Micellar electrokinetic chromatography (MEKC)............................... 408
14.2.3 Gel filled capillary electrophoresis(GFCE)........................................ 411
14.2.4 Capillary isoelectric focusing (CIEF).................................................. 412
14.2.5 Instrumentation ................................................................................... 412
14.2.6 Capillary.............................................................................................. 412
14.2.7 High voltage power supply.................................................................. 413
14.2.8 Temperature control ............................................................................ 413
14.2.9 Injection .............................................................................................. 414
14.2.10 Detection ............................................................................................. 415
14.2.10.1 UV detection ...................................................................... 416
14.2.10.2 Fluorescence, indirect fluorescence, laser-induced fluo-
rescence .............................................................................. 418
14.2.10.3 Amperometric detection, conductometric detection, MS
detection ............................................................................. 419
14.3 Applications of capillary electrophoresis.......................................................... 421
14.4 Conclusion......................................................................................................... 424
14.5 References ......................................................................................................... 425
Subject Index ............................................................................................................ 427

XV
Foreword
Although the spectacular development of gas chromatography in the 1950s

and 1960s is now a fading memory of a golden era, there are still advances being
made in apparatus, technique and applications. The petroleum industry makes
use of all the variants of chromatography as should be clear from the present
volume, but gas chromatography in particular is the most important analytical
technique in petroleum analysis and has been since its first announcement by
James and Martin in 1952. Indeed it is no exaggeration to claim that many of the
major advances in gas chromatography in that golden era emanated from the
laboratories of the petroleum industry.
This book is intended primarily for those concerned with the analysis of crude
oil and its products but many of the chapters have much broader applications. It
is hoped, therefore, that many outside the immediate sphere of petroleum analy-
sis will find sufficient of interest to make it a worthwhile purchase.
In multi-author books there will be inevitable variations in the style and con-
tent of each contribution. There is no reason why this should be regarded as a
weakness since as William Cowper pointed out varietys the very spice of life.
Likewise a small amount of overlap between some chapters is not a drawback if
it allows each chapter to be a freestanding account of a particular topic.
It was interesting to reread the comments of the editors of the only other book
dedicated to the subject of petroleum analysis by chromatography published
15 years ago. These editors spent some time describing the reasons for the
choice of the title of their book. In this context, it is interesting that the original
title intended for their book was the one used here.
In concluding this foreword, I should like to thank all the contributors and El-
sevier for their efforts to make this both a useful and an interesting volume.
E.R.Adlard
XVII
List of Contributors
D.J. ABBOTT Esso Research Centre, Analytical Group, Abingdon, Oxford-

shire OX13 6AE, UK
A. BARKER Dussek Campbell Ltd, Thames Road, Crayford, Dartford, Kent
DAI 4Q.J UK
J. BEENS Koninklijke/ShelI-Laboratorium, Amsterdam {Shell Research
B. K), Badhuisweg 3, 1031 CMAmsterdam, The Netherlands
G. BONDOUX Waters Chromatography Division, Millipore S.A., 6 Rue Jean-
Pierre Timbaud, BP 307, 78054 St. Quentin-en- yvelines,
France
J. BOS Koninkl~~/SheII-Laboratorium,Amsterdam {Shell Research
C.J. COWER 84, West Grove, Walton on Thames, Surrey KT12 5PD, UK
A. DE WIT KoninklijkdShell-Laboratorium, Amsterdam (Shell Research
N. DYSON @son Instruments Ltd, Hatton Lyons Industrial Estate, Hatton,
Houghton-le-Spring, Tyne and Wear DH5 Om, UK
R.S. HUTTE Sievers Instruments Inc., 1930 Central Avenue, Suite C, Boul-
der, CO 80301, USA
T. JONES Waters Chromatography Division, MiIlipore (CK) Ltd, Winster
House, Heronsway, Chester Business Park, Wrexham Road,
Chester CH4 SQR, UK
T.P. LYNCH Analytical & Applied Science Division, BP Research & Engi-
neering Centre, Sunbury-on-Thames, Middlesex, T W I 6 7LN
UK
H. MAHLER Siemens AG, Abt AUT 35 CHR, Postfach 21 12 62, 76187
Karlsruhe, Germany
T. MAURER Siemens AG, Abt AUT 35 CHR, Postfach 21 12 62, 76187
Karlsruhe, Germany
F. MUELLER Siemens AG, Abt AUT 35 CHR, Postfach 21 12 62, 76187
Karlsruhe, Germany
A.C. NEAL Esso Research Centre, Milton Hill, Abingdon, Oxfordshire
OX13 6AE. UK
XVIII List of contributors
1. ROBERTS Analytical & Applied Science Division, BP Research & Engi-

neering Centre, Sunbury-on-Thames, Middlesex, TW16 7LN,
UK
A.T. REVILL CSIRO Division of Oceanography, Castray Esplanade, Hobart,
Tasmania, Australia
S.J. ROWLAND Petroleum and Environmental Geochemistry Group, Depart-
ment of Environmental Sciences, Universily of Plymouth, Drake
Circus, Plymouth, PL4 8AA, UK
A. SIRONI Fisons Instruments, Strada Rivoltana, 20090 Rodano (Milan),
Italy
R. TIJSSEN KoninklijkdShell-Laboratorium, Amsterdam (Shell Research
B. V.), Badhuisweg 3, I03 I CM Amsterdam, The Netherlands
G.R. VERGA Fisons instruments, Strada Rivoltana, 20090 Rodano (Milan),
Italy
E.R. Adlard (Ed.), Chromatography in the Petroleum Industry
Journal of Chromatography Library Series, Vol. 56
0 1995 Elsevier Science B.V. All rights reserved 1
CHAPTER 1
The analysis of hydrocarbon gases
C.J. Cowper
British Gaspfc,London Research Station, Michael Road, London SW6 ZAD, UK
That man ... sat down to write a book, to tell the world what the world had all his life been
telling him.
Boswells Life of Johnson
1.1 INTRODUCTION
Hydrocarbon gases can be categorized in a number of ways, one of which is to

define them as natural or man-made.
Natural gas is a major energy source for domestic, commercial and industrial
consumers, and is used, so far as possible, with minimum change to the compo-
sition found in the reservoir. It consists generally of methane and other saturated
hydrocarbons and some non-flammable gases. Man-made gases arise from refin-
ing operations on liquid hydrocarbon feedstocks, and their compositions vary
widely according to the process from which they are derived. Those components
found in natural gas are likely to be present, in addition to unsaturated hydrocar-
bons.
Gas chromatography is the principal analytical method used for hydrocarbon
gases. Particular components can be measured by spectroscopic or chemical
means, but for analysis of the bulk of components, the separating power of
chromatography is both essential and well developed.
Although gases are often considered to be simple mixtures, their analysis has
frequently tested the ability of gas chromatography, either because of the range
of components present (boiling range, or concentration spread, or both), or be-
cause of the need to use highly specific stationary phases to separate apparently
intractable pairs of components. The different separating requirements relating
to groups of components within the same gas mixture has led to the use of multi-
column systems, with columns being isolated or reversed, or their order changed
* Current address: 84, West Grove, Walton on Thames, Surrey KT12 SPD, UK.
References p . 40
2 Chapter I
by means of valves. This complexity, which is more daunting in prospect than in

use, has led to a number of ready-configured chromatographic systems for many
of the application areas.
The thermal conductivity detector (TCD) and flame ionization detector (FID)
are the two most commonly used for hydrocarbon gases in the petroleum indus-
try. Because many of the gases contain non-hydrocarbon components, the TCD,
as a universal detector, is essential. Its dynamic range allows it to be used also
for all the major and many of the minor components of most mixtures. The FID,
while the most commonly used detector in gas chromatography generally, can be
regarded as a specialist and specific detector in gas analysis. Process chroma-
tographs frequently use the TCD alone, to reduce the need for the extra facilities
needed for the use of the FID.
1.2 NATURAL GAS
Hydrocarbon gases arise naturally from a variety of sources. Bacterial fermen-

tation under anaerobic conditions produces methane or marsh gas in great pro-
fusion, about 109 tonnes per year worldwide. Small accumulations of this type of
gas can be found during tunnelling or other operations, and the same mecha-
nisms produce landfill gas from waste. Mine drainage gas is a methane-rich
mixture found where coal measures have been worked. However, the term natu-
ral gas is normally taken to refer to the fossil-based gaseous equivalent to oil and
coal, abstracted from ancient, large, deeply buried accumulations. This is the
sense in which the term is used in this chapter.
Natural gases can vary considerably in composition, from nearly pure nitrogen
to nearly pure carbon dioxide to nearly pure methane. Fortunately for the indus-
try and the consumer, most natural gases consist mainly of methane, with small
amounts of inert gases (helium, nitrogen and carbon dioxide) and ethane and
higher alkanes in concentrations which diminish as their carbon number in-
creases.
By far the largest use of natural gas is as a fuel, where its accessibility via
wide-ranging distribution systems and its cleanness in terms of handling and
combustion products make it a popular choice for both domestic and industrial/
commercial markets. Other uses are as a chemical feedstock, as a source of pure
single hydrocarbon gases or (if present in sufficient quantities) of helium, and as
a moderator in nuclear reactors.
Current estimates indicate that the world has more reserves of natural gas than
of oil at the present rate of consumption. Recent measures of worldwide produc-
tion give a figure of around lo9 tonnes per year, which is comparable to the bac-
terial production referred to earlier.
The analysis of hydrocarbon gases 3
Natural gas is part of a continuum of hydrocarbons, ranging from methane to

the heaviest ends of oil, which are found in geological accumulations. Pressure
and temperature conditions in the reservoir are such that there is no distinction
between what we regard as gases and liquids; this only occurs when the fluid has
been extracted and is subject to conditions at which this discrimination is possi-
ble. Whether an accumulation is regarded as a gas or oil field is only a matter of
the relative proportions of the hydrocarbons. Natural gas fields always contain
liquids, usually in the form of a lightish condensate, and oil fields always contain
associated gases.
Gas separated from a natural gas field will burn in that form, but is usually
treated to remove or to control the levels of particular components, for opera-
tional, or contractual, or legislative reasons. Hydrogen sulphide, being toxic and
corrosive, is invariably subject to very low (parts per million) specification lim-
its, and is typically removed in an amine plant. Carbon dioxide is less acidic, but
still potentially corrosive at the pressures used for gas transmission, and its con-
centration is also controlled, usually to low percentage levels. It can be removed
by an alkali scrubbing process. Water is removed by glycol scrubbing, since the
presence of liquid water increases the corrosive effect of acid gases, and because
it can form solid methane hydrate, a clathrate compound, under certain pressure
and temperature conditions. Potential hydrocarbon liquids are also removed,
usually by chilling, sometimes by adsorption. This is to prevent their condensa-
tion downstream of the processing plant.
The fact that natural gas, once processed at the wellhead or reception termi-
nal, is in the form which virtually every consumer can accept without modifica-
tion has given rise to very complex and detailed pipeline systems, which cross
international boundaries and finally enter the consumers premises. In Western
Europe, most countries have access to pipeline supplies from Holland, the North
Sea, Siberia and Algeria in addition to their own indigenous sources. In the
United States, which is the home of long-distance natural gas transmission,
pipeline systems include Canada and Mexico as well as extensive offshore net-
works.
Properties and behaviour of natural gas have been reviewed by Melvin [ 13. A
large number of papers on quality specifications, physical properties, sampling,
odorization and analysis of natural gas, and on calibration gases and standardi-
zation are collected in the proceedings of the 1986 Gas Quality Congress [2].
Analysis of natural gas is carried out for a range of purposes, and the choice
of analytical method is often dictated by the reason for the analysis being re-
quired. There are three basic purposes for analysis:
- identification of source,
- calculation of physical properties, and
- measurement of specific minor components because of their particular
characteristics.
References p . 40
4 Chapter I
For identification of source, the concentrations of the inert components and

the ratios of a small number of hydrocarbons are good indicators; the analysis
need not be detailed. An example of specific minor component analysis is the
measurement of odorants; the analysis is clearly targeted upon a few compo-
nents, probably using a selective detector, and the composition of the main com-
ponents is without interest, except insofar as they may interfere with the meas-
urement. Calculation of properties is the most common need for analysis, with
calorific value the most usual target.
The following is a list of some of the properties of natural gas which are cal-
culable from analysis. It is not comprehensive, but describes those most fre-
quently used. Most properties can be measured directly, but independently of
each other; a properly configured analytical method allows calculation of all.
1. Culorijic value (CV): Natural gas is bought and sold in units of volume, as
a source of energy, hence the importance of CV as energy per unit of volume.
2. Relative density (RD): This is the density of a gas relative to dry air
(= 1.000). It is used in metering calculations and for the Wobbe index (see be-
low).
3. Wobbe index (WI): Gases from different sources must be assessed for their
inter-changeability, which represents the effectiveness with which a gas of com-
position B will burn on an appliance designed for a gas of composition A. WI is
an empirical measure of the ability to supply heat to a burner, and is the most
important characteristic in determining interchangeability. It is calculated by di-
viding the CV by the square root of the RD.
4. Compression factor (Z): Compression factor appears in the modified ideal
gas equation P V = nZRT, and arises from gas phase interactions. For hydro-
carbon gases and their mixtures over normal temperature and pressure ranges, Z
is always less than 1 , which means that a defined volume of gas at a defined
pressure will contain more moles than predicted from ideal behaviour by a factor
of 1/Z. At ambient conditions, Z for most natural gases is around 0.997, but the
correction is much more significant at higher pressures. At 70 bar, typical of
transmission pressures, Z is usually less than 0.9. Metering at high pressure is
therefore very dependent upon accurate measurement or calculation of Z.
5 . Hydrocarbon dewpoint: Retrograde condensation is the phenomenon
whereby a liquid phase can separate from a hydrocarbon gas mixture as it is de-
pressurized at a constant temperature. It is another feature of gas phase interac-
tions, and may be regarded as a form of gas phase solubility, with components
coming out of solution as the pressure binding the molecules together is re-
leased.
6. .Joule-Thomson coefficient: This property influences the extent of cooling
as a gas is expanded. As the pressure of natural gas is reduced, the amount of
pre-heating necessary to avoid hydrocarbon condensation can be calculated.
1.2.1 Analytical requirements

Although distributed natural gases consist mostly of methane, they are in fact
complex mixtures of a dominant major component (methane), a small number of
components in the range 0.1-15%, and a large number of trace components. Fig-
ure 1.1 shows the boiling points of both major and minor components from he-
lium to n-decane, indicating a boiling range of over 400C. While gases are not
often considered in terms of their boiling points, it is a good illustration of the
potential chromatographic problem. As a simple rule of thumb, an isothermal
separation will handle components with a boiling range of about 100C. The ap-
proach to natural gas analysis, therefore, can be to split it up into a series of
separations of groups of components, a temperature programming approach, or a
column switching method. Analytical needs are considered below in respect of
two important properties: CV and dewpoint calculation.
1.2.1.1CVmeasurement
The CV of a gas mixture is an additive property, with inert gases contributing
zero, and flammable gases contributing in proportion to their concentration and
individual CV. A small correction is necessary for compression factor (2)at
ambient conditions.
Figure 1.2 shows, for a typical North Sea gas, the component contributions in
terms of molar %, and of CV and RD as percentages of the total. Nitrogen, pres-
ent at 2.5%, contributes nothing to the CV, but 4% to the RD. The Y-axis of the
figure is limited to 6% so that component contributions can be clearly seen.
Methane, of course, contributes far more than the figure indicates.
B.R. deg C
200 I I
-300 I 1
He N2 0 2 C02 C1 C2 C3 C4 C5 C6 C7 C8 C9 C10
Component
Fig. 1. I . Component boiling points.
References p . 40
6 Chapter I
% Contribution
N2 C02 C1 C2 C3 164 nC4 nee= IC5 n-C5 C6+
Component
I M o i a r % B C a l Value =Re1 Density
Fig. 1.2. Component contributions.
Figure 1.3 expands the information for the higher hydrocarbons (C, and
above). It is clear that the relative contribution to CV and RD is greater with in-
creasing carbon number, but the diminishing concentrations means that the ac-
% contribution
0.2 17
0.15
0.1
0.05
0
nC5 C6 Benr cyC6 C7 To1 MecyC6 CB C9 C10
Component
mMolar % BCal Value B R e l Denslty
Fig. 1.3. Component contributions.
The analysis ofhydrocarbon gases 7
Fig. 1.4. Calorific value errors.
tual contribution is small. Figure 1.4 shows the error involved in MJ/m3 if a
component or group of components is missed. (The total CV would be around
38 MJ/m3). This is shown for components or groups, and also cumulatively, from
a particular carbon number upwards. Thus, if the Clo hydrocarbons are not
measured, but their molar contribution is assumed to be included with that of
methane, the CV will be underestimated by only 0.003 MJ/m3. C, hydrocarbons,
if not measured, would cause an underestimate of 0.009 and C, hydrocarbons of
0.015 MJ/m3. It is, of course, much more likely that if the C, hydrocarbons have
been missed, so also would the C, and C,, hydrocarbons, giving a cumulative
error. In this case, missing C, and above would give an underestimate of
0.027 MJ/m3, and if C, and all higher hydrocarbons are not measured, the error
will be about 0.075 MJ/m3.
A calculation uncertainty of 0.1 MJ/m3 is a realistic target for a properly con-
figured and accurately calibrated analyser, and so to minimize the bias error
arising from undetermined components, the analysis should include C, and pref-
erably C, hydrocarbons. One of the common methods of analysis backflushes all
C, and higher hydrocarbons to the detector, where they are measured as a com-
posite C,+ peak. This includes all higher hydrocarbons in the measurement, but
raises two further uncertainties: unless there is independent information about
the detailed composition of this peak, its response factor must be relatively ill-
References p . 40
x Chapter 1
defined, and so must its contribution to CV or other properties. In fact, the CV of

the C,+ fraction of many gases can be approximated by that of n-hexane without
significant error. Components such as benzene and toluene, and to a lesser extent
the cyclo-alkanes have lower CVs than alkanes of equivalent carbon number,
and if present in reasonable proportion can counteract the higher CV contribu-
tions of C, and higher alkanes.
1.2. I 2 lfydrocarbon dewpoint calculation
Calculation of hydrocarbon dewpoint temperature is complex, as interactions
between components must be accounted for in addition to individual component
properties, Higher hydrocarbons make a considerable contribution, because of
their relatively low vapour pressures. For CV calculation, it is normal to con-
sider all alkanes of a particular carbon number as a group. Since the CVs of al-
kane isomers are very similar, this is realistic and involves virtually no loss of
accuracy. The same approach is incorrect for hydrocarbon dewpoint calculation,
as the contributions of isomers differ. This creates two problems: computer
packages for these calculations cannot handle as many components as a detailed
analysis can measure, and even the most detailed analysis cannot definitely
identify all the peaks which it separates, nor find the appropriate properties for
those components through a database.
A typical computer program will handle 30 components, and one approach
has been to group alkane isomers as if their sum was represented by the n-alkane
of that carbon number. Since the n-alkane has the highest boiling point of the
series, this approach will over-estimate the contributions to dewpoint tempera-
ture, and so has the advantage of a built-in safety margin. A more accurate ap-
proach is to input data for groups of components as that of fractions rather than
components, assuming that the program allows components and fractions to be
mixed.
Detailed separation of higher hydrocarbons is most likely to be on the basis of
boiling point, as in simulated distillation. Each peak in the chromatogram, with-
out being identified, can have a boiling point allocated to it based upon its reten-
tion time relative to bracketing n-alkanes, a carbon number based upon its posi-
tion in the chromatogram, and hence an FID response factor and molar percent-
age. It is therefore practicable to consider a group of hydrocarbons, such as the
C, alkanes, not as n-C, but as the C, fraction. This fraction has a defined molecu-
lar weight and density, a molar concentration and a calculated average boiling
point. This is sufficient information to be able to input the C, data as a fraction
with properties which more realistically represent its contribution. The same ap-
proach can be used for C,, C,, C,, and any higher hydrocarbon groups which
may be measured.
Figure 1.5 shows the errors involved in dewpoint temperature calculation if
components or groups of components are not measured, and their molar contri-
The analysis of hydrocarbongases 9
Fig. 1.5. Dewpoint temperature errors.
bution taken up by that of methane. The failure to measure individual compo-

nents or groups produces errors measured on the left hand scale. The significant
role of the higher hydrocarbons is clear, with a C,, error of -3.7"C and a C, error
of -5C. By contrast, ignoring propane will only give a -0.4"C error, and ignor-
ing ethane or the inert components hardly any.
As with the CV errors above, it is likely that if C, is missed from the analysis,
so also will Clo. This cumulative error, where failure to measure hydrocarbons
of any carbon number also includes those of higher carbon number, is shown
with reference to the right hand scale, where the range is 10 times greater. It is
evident that the cumulative errors are larger than would be assumed by addition
of the individual contributions. Failure to measure C, and higher components
will cause an underestimate in calculated dewpoint temperature of more than
50C.
Analysis for dewpoint calculation must, therefore give details of higher hy-
drocarbons. The example is for a gas where C,, and higher components are not
detectably present, which is normal for gases treated to a transmission specifica-
tion. If they were detectably present, they must be measured and included in the
calculation. While it appears that the analysis need not measure lighter compo-
nents particularly well, if at all, it would be bizarre and unusual to configure an
analytical scheme to be poor for the easy to separate light components and good
for the more difficult traces of heavy ones. Also, a particular analytical result can
References p . 40
10 Chapter I
be and frequently is used for calculation of several physical properties, which

requires high quality of measurement across the range.
1.2.2 Analytical procedures

Considering the range of components in Fig. 1.1, there are several incom-
patibilities which will influence the choice of method. Helium is the preferred
carrier gas for TCD measurement of most components, and so cannot be in-
cluded as a component during measurement of the others. Oxygednitrogen sepa-
ration requires the use of a molecular sieve column (unless sub-ambient tempera-
tures are used), which is incompatible with measurement of CO,. The range of
component boiling points requires different columns or temperature program-
ming.
1.2.2.I Isothermal methods
IP 337 [3] recommended the use of three different separations, a molecular
sieve column with argon carrier gas for He, 0, and N,, a porous polymer column
with helium carrier operated at 50C for CO, and C, and a porous polymer col-
umn with helium carrier operated at 140C for C,, C, and C, hydrocarbons The
analysis went no further than C,, and methane was measured by difference.
ASTM D 1945 [4] sought to achieve measurement of more components in a
single separation, which included a 10-m column with a high loading of silicone
oil on Chromosorb P. This separated N,, C,, CO, and C, to C, hydrocarbons in-
dividually. A molecular sieve column was still necessary for measuring air com-
ponents.
Figure 1.6 shows a separation on a boiling-point column (6 m of 28% silicone
oil DC 200/500 on Chromosorb P at 100C). C, C, and C, hydrocarbons are
well separated, but the light components, N,, C,, CO, and C,, while distinct, are
not sufficiently well separated for best quantitative measurement. In particular,
CO, at low concentration can be difficult to discern between C, and C,. Hydro-
carbons above C, are slow to elute, and the combination of their decreasing con-
centrations and increasing peak widths makes their measurement more and more
difficult. (In this and subsequent figures, only the n-alkanes are identified, to
avoid clutter).
Backflushing to recombine all hydrocarbons above C, and pass the combined
peak (C,+) to the detector has two advantages: the recombined peak will be
larger than the individual ones, and the analysis time will be reduced. Against
this, we cannot make exact allowance for the contribution of all the individual
components, but must make some assumptions about the bulk properties. Figure
1.7 shows the valve system which allows rapid backflushing and measurement of
C,i. The original boiling point column is separated into a short (0.75 m) and a
long (5.25 m) section. The valve both alters the sequence of these sections, and
fAlten.
I I I
0 min 10 min 20 min
Fig. 1.6. Boiling point separation. Column: 6.0 m X 2 mm id., 28% DC 200/500 on 45/60 mesh
Chromosorb P-AW. Temperature: 100C. Carrier gas: helium at 28 ml/min.
the direction of carrier gas flow through the short section. The column lengths
are chosen so that after injection (Fig. 1.7a), n-C, will have passed into the
longer section before the lightest component has emerged from it. Reversing the
valve after n-C, has passed this point (Fig. 1.7b), a time which is found by trial
and error, causes all higher hydrocarbons to recombine and emerge from the
short column as a sharp C,+ peak. The normal, forward eluted components then
follow, as shown in Fig. 1.8. After measurement of n-C,, the system is immedi-
ately ready for another analysis. Figure 1.7 shows the use of a single 10-port
valve for both sample injection and backflushing. If preferred, two six-port
valves, one for injection, one for backflush, may be used instead.
Figure 1.6 illustrates the problem with the wide boiling range of the mixture.
C, to n-C, (boiling range 78C) are well separated in a reasonable time, while
the lighter gases are somewhat overlapped and the C, and heavier components
are slow to elute and difficult to detect. Backflushing of C,+ solves that part of
the problem, although introducing uncertainty about composition; the lighter
gases need different conditions for good separation. Since they emerge rapidly
and in a group, it is possible to divert them to a separate column, more suitable
for their separation, and then to allow the C, to C, components to emerge and be
detected as before. A porous polymer bead column will give good separation of
References p. 40
12 Chapter I
a Sample
2
1,
--<
ez j:
1
I
Detector
d
Column
1
b Sample
Carrier
._ v1 I
!
I
Gas \ J l
1 -.- I
Fig 1.7. Accelerated backflush: (a) valve 1. position 1, inject and forward flow; (b) valve 1. posi-
tion I . sample load and backflush.
these light gases at the same temperature as the boiling point column uses for the
C3 to C, separation.
Figure 1.9 shows the configuration which achieves this, with valve 2 serving
to include or isolate column 3 , the porous polymer column. Restrictor A is ad-
justed to give the same pneumatic resistance as column 3 , so that the carrier gas
The analysis of hydrocarbon gases I3
I
0 min 10 min 20 min
Fig. 1.8. Boiling point separation with backflush. Column 1: 0.75 m X 2 mm i.d. Column 2:
5.25 m X 2 mm i.d. Both containing 28% DC 200/500 on 45/60 mesh Chromosorb P-AW. Tem-
perature: 100C. Carrier gas: helium at 28 ml/min.
flow remains constant. With column 3 in series, the sample is injected via valve
1. As before, C,+ is backflushed to the detector by returning valve 1 to the load
position. As soon as all the C, has passed into column 3 (found by trial and er-
ror), valve 2 is switched to isolate the light gases, N,, CO,, C, and C, in that col-
Sample
Column 3
I I
Carrier
Gas
WLoop
\ I
,
I
I
I
I
Restrictor A
I
7-
Column 2
Fig. 1.9. Three column analyser. Valve 1, position 2; valve 2, position 1
Referencesp . 40
14 Chapter I
0 min 10 min 20 min
Fig. 1.10. Boiling point and polymer bead column. Column 1: 0.75 m X 2 mm i.d. Column 2:
5.25 m X 2 mm i.d. Both containing 28% DC 200/500 on 45/60 mesh Chromosorb P-AW. Col-
umn 3: 2.4 m x 2 mm i.d. 15% DC 200/500 on SOB0 mesh Hayesep N. Temperature: 100C. Car-
rier gas: helium at 28 ml/min.
umn. C,, C, and C, hydrocarbons emerge from columns 2 and 1 to the detector.
After n-C, has eluted, column 3 is returned on-line by switching valve 2, and the
light gases elute and are measured. Figure 1.10 shows a typical chromatogram.
The above configuration, with possible minor variations, is widely used for
on-line natural gas analysers, where the sample stream is connected in such a
way that the possibility of contamination of the sample by air is minimal. How-
ever, any sample returned for analysis to a laboratory is prone to air contamina-
tion, which means that 0, and N, must be separated if the presence of air is to be
recognized, and accurately measured if the air-free composition is to be recalcu-
lated. 0, may also be present in a transmitted natural gas if air or N, ballasting is
used as a means of controlling CV or WI.
Porous polymer bead columns will not separate air components at the tem-
peratures used for normal analysis; molecular sieves are the only materials able
to do this, and they in turn retain CO, for so long as to make it unmeasurable.
The solution then is to cut the N, and CH,, with any 0, which may be present,
onto a molecular sieve column, leaving the CO, and C, on the porous polymer.
Figure 1.1 1 shows the configuration, where the additional valve 3 includes or
isolates the molecular sieve.
The procedure is similar to the previous one, starting with all columns in se-
ries. After backflush of C,+, the light gases pass into the polymer bead column,
Sampla Column 3 Column 4
Carrlrr
Gas
Restrlctor A ( I Reitrlctor B
I
Column 2
Fig. 1.11. Four column analyser. Valve 1, position 2; valve 2, position 1; valve 3, position 1.
but now the N, and CH, (and 0, if present) are allowed to go forward to the
molecular sieve column. Both columns are now isolated, with CO, and C, in
column 3, the porous polymer, and 0,, N, and CH, in column 4,the molecular
sieve. After elution of C,, C, and C, hydrocarbons, column 3 is reconnected for
elution and measurement of CO, and C,, and finally column 4 is connected for
measurement of 0,, N, and CH,. Figure 1.12 is a typical chromatogram.
c3 c2 N2 CH4
I I I
0 min 10 min 20 min
Fig. 1.12. Boiling point, polymer bead and molecular sieve. Column 1: 0.75 m X 2 mm i.d. Col-
umn 2: 5.25 m x 2 mm i.d. Both containing 28% DC 200/500 on 45/60 mesh Chromosorb P-AW.
Column 3: 2.4 m x 2 m m i.d. 15% DC 200/500 on 50/80 mesh Hayesep N. Column 4:
2.4 m x 2 mm i.d. 45/60 mesh Molecular Sieve 5A. Temperature: 100C. Carrier gas: helium at
28 ml/min.
Re$rences p. 40
16 Chapter 1
The above chromatograms were generated, as is evident from the conditions,

on a single chromatograph, configured as in Fig. 1.1 I . The configuration of Fig.
1.9 (chromatogram in Fig. 1.10) was achieved by isolating column 4 throughout,
and that of Fig. 1.7 (chromatogram in Fig. 1.8) by isolating columns 3 and 4
throughout. It would be possible to optimize columns for these configurations
which gave somewhat better N,/CH, separation, but it is clear that the baseline
separations of all light components seen in Fig. 1.12 could not be matched.
The setting-up procedures described above sound more complicated than they
in fact are and most instrument suppliers provide ready configured systems.
Once set up, their performance is usually extremely stable, since none of the
coiumns are used anywhere near their temperature limit. The molecular sieve
column will lose separation gradually due to slow adsorption of moisture, but
its performance does not influence other timings or separations within the sys-
tem.
1.2.2.2 Two detectors

The chromatogram in Fig. 1.12 illustrates the use of two detectors, TCD and
FID in series. The presence of N,and CO, makes the use of the TCD necessary,
and it is sufficiently sensitive for measurement of the lighter hydrocarbons. To
include pentanes, however, means choosing a larger sample size than would be
desirable for linear measurement of the major components. Using an FID in
series with the TCD avoids this problem, as the FID has much higher sensitiv-
ity for hydrocarbons. A smaller sample size can be used, satisfying linear detec-
tion requirements for major components and ample sensitivity for minor compo-
n ents .
1.2.2.3 C 6 detail
~
Separation and measurement of the individual higher hydrocarbons, repre-
sented by the C,+ backflushed peak, is needed both to define the composition
and hence the properties of this group, and also to provide detail for other calcu-
lations such as hydrocarbon dewpoint temperature. The complexity of the minor
alkane isomer components increases dramatically with carbon number, and this,
with the presence of cyclo-alkanes and aromatics, means that high resolution
chromatography is required.
Capillary columns are widely used for liquid hydrocarbon samples, and are
equally adaptable to gas analysis. The sample injected from a conventional gas
sampling valve can be split without fear of sample discrimination, as there is no
phase change on injection. Alternatively, the capillary column can be connected
directly into a micro-volume gas sampling valve, fitted with a sample loop of
some tens of microlitres. Chromatograms of natural gases with very different
isomer distributions are shown in Figs. 1.13 and 1.14. The limit of detection for
individual components in this instance is around 5 parts per million molar.
I
0 min i0'min 20 rnin tomin
Fig. 1.13. Capillary separation.
I I
0 min 10 min 20'min 3Omin
Fig. 1.14. Capillary separation. Column: 50 m X 0.2 mm i.d. capillary coated with OV-101. Tem-
perature: 35C for 5 min, then 6"C/min to 220C. Carrier gas: helium at 1.3 bar.
Referencesp. 40
18 Chapter I
I . 2.2.4 Temperacure programming

Figure 1.6 shows a chromatogram with good resolution and peak shapes for
C, to C, hydrocarbons, insufficient resolution for lighter components and long
analysis time and low, broad peaks for C, and heavier components. The solution
consisted of diverting groups of components onto different columns, chosen to
optimize their separation at the selected operating temperature. Another ap-
proach is to extend the range by programming the temperature of a single col-
umn.
The boiling point column used for Fig. 1.6 would only give good separations
of the light gases by using a low sub-ambient column temperature, and so
Stufkens and Bogaard [ 5 ] proposed the use of a porous polymer bead column,
which has inherently longer retentions. They chose Porapak R, a material with
intermediate polarity which optimizes the separation of C,, CO, and C,. Using a
-.IO"C to 230C programme, N,, CO, and C, to C, hydrocarbons are well sepa-
rated for individual measurement, and C,, C, and C, hydrocarbons are measured
as groups. n-C, elutes at the upper temperature limit for the column, and any
heavier components cannot feasibly be measured. A TCD and an FID were used
in series, with ethane acting as a bridge component to link the detector re-
sponses. Methane was measured by difference, although there is no reason
why it could not be measured directly. The method is intended for CV determi-
nation.
I
0 min 12 min 24 min
Fig. 1.15 Temperature programmed separation. Column: 3 m X 2 mm i.d. 50180 mesh Porapak R.
Temperature: -50C for 2 min, then ISWmin to 240C. Carrier gas: helium at 30 ml/min.
Using this method, but with a sub-ambient (-50C) start to the temperature
programme, O,/N, separation is achieved within the same analysis. Figure 1.15
is a typical chromatogram. I S 0 6974 [6] is based on this separation, with a
starting temperature of 35"C, which eliminates the need for sub-ambient equip-
ment, but an additional separation on a molecular sieve column is required for
helium, oxygen and nitrogen.
1.2.2.5 Combined systems
The capillary chromatogram in Fig. 1.13 separates the majority of components
in a natural gas, but not those few lighter components which have the largest
concentrations. Figure 1.13 has been optimized for good detection limits over a
wide range, giving quantitative measurement from C, to C,, and beyond. Used in
conjunction with the column switching system illustrated in Fig. 1.11, either C,
or C , components can be used as a quantitative bridge between the two analyses.
If the capillary conditions are optimized in order to include quantitative meas-
urement of C,, then the packed column TCD system for light gases can be sim-
plified to a porous polymer/molecular sieve combination. Both this and the capil-
lary/FID system can be fitted into the same chromatograph [7]. The sample is
injected first into the packed column system for isothermal separation of 0,, N,,
C,, CO, and C,, and then into the capillary for temperature programmed separa-
tion of the higher hydrocarbons. There is no backflush provision to remove the
heavier hydrocarbons from the packed columns, as they are forward eluted (but
not measured) during the temperature programmed part of the cycle. Figure 1.16
illustrates the column arrangement and Fig. 1.17 a typical chromatogram.
The analysis is comprehensive, but there is no provision for a bridge compo-
nent. C, and C, can be distinguished on the capillary column, but it is doubtful
whether C, could be measured with sufficient accuracy. Although the analysis
takes 30 min, the cycle time, allowing for cool down and re-stabilization, is
longer. In a similar application, three sample aliquots are injected, one onto mo-
lecular sieve for O,/N, separation, and from which C, and above are backflushed
to vent, one onto porous polymer for (0,+ N,), C,, CO, and C,, and the third
onto a capillary for temperature programmed separation [81.
1.2.2.6Separation in backflush
In a novel application intended for process use, the natural gas is injected onto
a single, long packed column containing porous polymer and operating with a
large pressure drop. N,, C, and CO, are measured normally, then the entire col-
umn is backflushed to the detector for measurement of the other components. To
understand how this works requires careful consideration of the mechanism of
backflushing in gas chromatography.
Backflushing is assumed to recombine separated or partially separated com-
ponents because they have to travel equally far in the reverse direction from that
References p. 40
20 Chapter 1
Sample Molecular sieve
Capillary column (W
Fig. i 16. Combined system.
CH4 c4 c5 c6
- - _____
I I
0 min 15 min 30 min
Fig 1 17 Combined s> stem chromatogram.
The analysis of hydrocarbongases 21
in which they have been undergoing separation. This is an over-simplification. In

gas chromatography, the mobile phase is compressible, and is driven through the
column under the influence of a pressure gradient. Since each section of the col-
umn has the same mass flow of mobile phase passing through it at any time, the
actual volumetric flow at each point must vary according to the pressure at that
point. As the pressure decays from the beginning to the end of the column, so the
volumetric flow rate increases, and so does the linear velocity.
The mobile phase linear velocity controls the rate of travel of components,
and so if this increases along the length of the column, components, even in iso-
thermal analysis, will accelerate between injection and detection. Looked at an-
other way, a component with a retention time of x will not have reached the mid
point of the column at an elapsed time of x/2. The larger the pressure gradient,
the greater the acceleration.
When the whole column is backflushed, the pressure gradient which exists in
forward flow is reversed, and so those parts of the column where the components
travelled most slowly become the high velocity areas, and vice versa. A compo-
nent which has nearly reached the end of the column at the time of backflushing
will have experienced the full acceleration. It now finds itself in the high pres-
sure, low velocity region, and has to do it all over again. Its time to elute in
backflush will be almost identical to its forward flow time. By contrast, a com-
ponent which had travelled only a short distance from the injection point by the
time of backflushing will have spent all the forward flow time in the low veloc-
ity region. After backflushing, it is in the high velocity region, and will be eluted
as a backflushed peak in a much shorter time than it spent travelling forward.
Hence there is a mechanism of separation, in reversed order, during back-
flushing. The fact that this is not evident when backflushing in applications such
as that illustrated in Figs. 1.10 and 1.12 is because in these, the backflushed sec-
tion is short. Hence, while it is subject to high pressure, low velocity on forward
flow, and vice versa on backflush, the pressure drop across the section itself is
small, and backflush separation does not occur. Figure 1.18 is a chromatogram
showing the application [9]. The advantages of this approach are that it requires
less hardware (a single ten-port valve would cover both injection and backflush-
ing) and the column can be long enough to give good N,, CH,, CO, separation
without a consequent time penalty for the heavier components. The main disad-
vantage is to do with the nominal C,+ group. Since the backflush separation is
good enough to separate C, from C, and C, from C,, it is almost certainly per-
forming some separation of C,, C, and C,. This being so, the C,+ group, which is
a sharp, distinct peak in Fig. 1.11, becomes relatively smeared out and more
difficult to quantify.
I . 2.2.7 Rapid analysis
Process natural gas analysers have analysis cycle times typically in the range
References p . 40
22 Chapter I
0 min 4 min 8 hin

Fig. 1.18. Backflush chromatogram. Column: 6 . I m X 0.8 mm i d . 20% DC-200 on 100/120 mesh
Chromosorb P plus 2.1 m x 0.8 mm i.d. 1001120 mesh Porapak N. Temperature: 65C.Carrier
gas: helium at 8 mlimin: inlet pressure 9.5 bar.
of 5-1 5 rnin. Laboratory analysers, which usually provide more information,

tend to be in the range of 25-45 min. A recent development in miniaturization of
chromatographic hardware [ 10,11] allows analysis times in the region of 30 s.
Normal monitoring of a natural gas pipeline is unlikely to require significantly
faster analysis times than conventional equipment provides. There are particular
applications, associated with natural gas treatment, however, where rapid re-
sponse is important. The other advantage of a rapid analyser is that it can pro-
vide ten or more analyses during the time that a conventional unit takes to com-
plete one. The mean of the ten results is statistically superior to a single result,
provided that the rapid analyser has comparable precision on a single analysis
basis.
The hardware uses techniques derived from semiconductor technology, with
the carrier gas and sample channels, the injection device and a miniature TCD
etched into the surface of a silicon wafer. Short, small-bore capillary columns
are bonded to the wafer, and the whole is fitted into a heated enclosure, with
carrier gas provided in the conventional manner. The basic chromatographic
module is about the size of a packet of cigarettes, and as normally provided, two
such units are fitted into a single, portable case. The TCD is the only detector
available at present, but it has been shown that with sufficiently low dead vol-
ume, the TCD can overtake the FID in sensitivity [12]. Injection is achieved by
pressurizing the sample to above carrier gas pressure, then opening a miniature
The anafysisof hydrocarbongases 23
diaphragm valve between the two for a defined time; sample size is adjusted by
selecting a different opening time.
An application is provided for natural gas analysis [ 131. One unit is fitted with
a small-bore capillary column (4m X 0.1 mm i.d.) with a thin film of non-polar
phase. The other has a 25 cm X 0.5 mm micro-packed column, containing po-
rous polymer beads. Sample is injected simultaneously onto both units. The po-
rous polymer separates N,, C,, CO, and C,, and the boiling-point column sepa-
rates C,, C, and C, hydrocarbons. Both separations take less than 20 s, and are
illustrated in Figs. 1.19 and 1.20.
The precision of analysis is comparable to that which is available from both
laboratory and process analysers. The response functions to different compo-
nents, which measure the extent to which the instrument is linear, are also com-
parable to those for conventional equipment. The construction of the equipment
does not allow options such as backflushing or column switching and isolation;
all components must be measured by forward elution. Although the temperature
of each module can be changed rapidly, temperature programming is not an op-
tion; the cool-down and restabilization time would nullify the benefits of rapid
analysis.
To extend the component range, the boiling-point column can be operated at a
higher temperature. It is possible to choose a temperature which still allows
quantitative measurement of C, on the tail of the C,/C, peak, and measure up to
n-C, in 20 s, or up to n-C, in 80 s (Fig. 1.21).
I
0 s0cs
Fig. 1.19. Rapid analysis, light gases.
References p. 40
24 Chapter I
r 1
I
0 Secs 20 secs
Fig. 1.20. Rapid analysis, C3 to C5 gases
0 sees 80 secs
Fig. 1.2 1. Rapid analysis, analysis to C8.

The analysis ofhydrocarbon gases 25
1.2.3 Quantitative measurement

Natural gas analysis has probably received more attention than any other type
of sample in respect of quantitative measurement. This is mainly in respect of
CV determination, because of the very large financial implications of the huge
volumes of natural gas which are traded. Other properties, important for engi-
neering or safety reasons, can be derived from the same set of composition data.
To be able to quote the value of a property with a defined level of uncertainty
requires that the entire procedure be considered, from sampling through calibra-
tion and analysis to calculation of composition, and thence to calculation of
properties, using the correct procedure and accurate base data on pure com-
pounds. Kenter et al. [ 141 have described the uncertainty calculations for natural
gas CV measurement.
Considering the analytical procedure alone, the following criteria must be
met.
1. It must be comprehensive, with no significant components or groups ig-
nored. Detailed judgement as to what is significant depends upon the appli-
cation.
2. Unless only intended for on-line use, it should recognize air contamination
of the sample.
3. All significant components or groups should be resolved so that there is no
mutual interference.
4. It must be precise.
5. The response for each component must be a consistent and predictable
function of concentration.
The requirement for accuracy is not included here, as the accuracy of chroma-
tographic analysis is principally determined by the accuracy of the calibration
standards used. The analytical method can, if the above criteria are satisfied,
transfer that accuracy to the sample. IS0 10723 [15], currently at committee
draft stage, describes tests of analyser performance broadly covering the above
headings.
1.2.3.I Comprehensive analysis
The analytical requirement, and further details such as the desired frequency
of analysis and the availability of equipment will allow the user to choose from
the options described above.
1.2.3.2 Air contamination
Air can contaminate a sample when it is taken into a container or when it is
subsequently transferred to the analyser, or both. If the analysis requires differ-
ent injections of sample for the measurement of different groups of components,
air contamination may be present in some injections and not in others, and may
References p . 40
26 Chapter 1
or may not be recognized. Wherever possible, samples subject to air contamina-

tion should be analysed using a method in which both air and normal natural gas
components are measured from the same injection.
1.2.3.3 Resolution
The ideal of baseline resolution is not possible for all components in a com-
plex mixture such as natural gas. Components to C, can be effectively fully re-
solved with packed columns, and most C , and C, isomers separated on a capil-
lary. Again the degree of resolution required for minor components depends on
the application.
I . 2.3.4 Precision
The approach to measuring the purity of a nominally pure substance is to
measure the impurities, and calculate the concentration of the substance by dif-
ference. Since methane is the dominant component in natural gas, one approach
has been to measure all other components and to calculate the methane by differ-
ence. Taking this approach, however, means that the calculated methane value is
also the sink for the errors involved in the determination of all the other compo-
nents. It is therefore preferable to measure all components, including methane.
Errors in chromatographic measurement can be regarded as being of two
types; those which affect each component differently, and those which influence
all components, or a substantial group of components to the same extent. An ex-
ample of the former is the effect of random electrical baseline noise on the deci-
sion as to where peaks begin and end. An example of the latter is variation in
sample size or a change in detector sensitivity. Sample size variation is quite
common in gas analysis, where the injection device uses a defined volume, but
the molar injected quantity depends upon the pressure and temperature of the
sample at the time of injection.
If all components are measured from a single injection, then sample size
variation has the same proportional influence on all of them. If they are meas-
ured with a single detector, then the results, which should add up to loo%, can
be normalized to that value provided that the necessary correction is quite small.
It is quite common to normalize data which add to between, for example, 98 and
102%, but to investigate larger variations. Different detectors, even if used in
series on the same column system, can and do show changes in sensitivity of
different magnitudes and sometimes in different directions. Consider the popular
combination of TCD and FID; the FID sensitivity depends, among other factors,
upon the carrier gas to hydrogen ratio, whereas the TCD is quite indifferent to
the FID hydrogen flow. It is common to find that normalization of data from the
two detectors gives results which are more precise for those components meas-
ured by TCD, but less precise for the FID. The TCD, even if only being used to
measure Nz, CO,, C, and Cz, accounts for those components which comprise the
great majority of the molar quantity, and hence are most influential on the nor-
malization correction. The FID may be measuring many more components, but
their total is small and so, therefore, is their influence on normalization.
For this reason, where two detectors are used, a bridge component should be
selected which gives a good and interference-free signal with each detector
[5,16]. The ratio of detector responses for this component is then measured at
each analysis and compared with the ratio that was found at the time of calibra-
tion. Small deviations in relative detector response can then be allowed for by
adjusting the data from the second detector.
This approach can also be used if the analysis is split up into different parts,
some components being measured on one analyser and some on another. The
bridge component, measured on each, is used to allow for changes in the re-
sponse of the different instruments.
1.2.3.5 Responsefunction
Because most chromatographs give a larger signal for what is reckoned to be a
larger concentration of component, it is convenient and comforting to assume
that the response to that component is linear, i.e. that it is represented by a
straight line through the origin when plotting instrument response against com-
ponent concentration. This assumption means that the response factor is inde-
pendent of concentration, and justifies calibration with one mixture, at a single
point. Unfortunately, the assumption is not necessarily true, particularly for
methane, the inevitable major component.
Chromatography consists of a series of compromises, including the choice of
sample size in order to allow detection of minor components and simultaneous
quantitative measurement of major components. By using a series of test gases,
of carefully selected and accurately defined compositions, response functions
can be evaluated and curves plotted. It is common to find that methane shows
significant deviations from linear behaviour, and other components, such as ni-
trogen and ethane, show lesser but still noticeable deviations. These deviations
will give rise to bias errors if they are not allowed for, with the size of the errors
depending upon the size of the deviations and upon the concentration differences
between calibration standard and sample.
Having quantified the component response function, typically as a polynomial
expression, these coefficients can then be used in place of the assumption of lin-
ear response for subsequent quantitative measurements of samples. There re-
mains the uncertainty of the long-term stability of these functions, and hence the
frequency with which they should be re-checked. If possible, a preferable course
would be to adjust the analytical conditions in order to ensure that linear re-
sponses are available for all components.
Deviations from linear response are of more concern for the TCD than for the
FID. This is not to suggest that the FID is free from such problems, but that the
Referencesp . 40
28 Chapter 1
TCD is more likely to be used for measurement of major components. The TCD
is an admirably simple device, and deviations from linearity should only occur
when the concentration of component in carrier gas within the detector exceeds a
certain value. Above this concentration, the incremental signal increase for an
increment of extra concentration diminishes, and the response, which had been
following a straight line, starts to curve.
This overload effect is only dependent upon the instantaneous concentration
of component in the detector. For the same amount of component, a peak that is
narrow and tall may give a non-linear response, whereas one that is broad and
shallow will not. As has been stated, methane is both the most likely candidate
and the most frequently observed component to give non-linear response. At the
same time, the different tactics used in isothermal analysis have a greater effect
upon the peak shape for methane than for other components, and hence will be
likely to influence the nature of its response. When eluted directly to the detec-
tor, as in Figs. 1.6 and 1.8, the peak is narrow and tall, promoting non-linear be-
haviour. Intermediate storage in and elution from a porous polymer column, as in
Fig. 1.10, produces a wider and less tall peak. Figure 1.12 shows the even wider
methane peak which results from storage on a molecular sieve, which should be
the most favourable in terms of linearity of response.
It is therefore evident that the benefit of the more complex configuration when
moving from Fig. 1.7 to Fig. 1.9 to Fig. 1.11 is not only extra detail and better
separation, but also improved linearity of response. The penalty is the extra time
required for the analysis.
1.3 REFINERY GAS
The refining of crude oil starts with distillation, converting the crude into a
series of fractions which will themselves form products or feedstocks. The light-
est fraction consists of propane and butanes, with small amounts of ethane and
pentanes. Subject to further separation, this forms product streams of propane
(typically 95% pure) and butane (typically 40% isobutane and 60% n-butane),
generally referred to as liquefied petroleum gas or LPG. The ethane content is
too low to form a useful feedstock, and is used as fuel gas around the plant.
The light gases can also be fed to the catalytic cracker, where the simple
mixture of saturated hydrocarbons is converted to a more complex mixture
which also contains unsaturates and inorganic gases. The more reactive unsatu-
rated components are the key to further processes, such as polymerization and
the production of oxygenates for gasoline.
Refinery gas is the name given to this catalytic cracker product, and it con-
tains hydrogen, oxygen, nitrogen, carbon monoxide, carbon dioxide, hydrogen
sulphide, and saturated and unsaturated hydrocarbons in the range C, to C,. It
may contain small amounts of higher hydrocarbons, such as C,, and may also
pick up, during the course of the processes for which it is used, traces of aromat-
ics or other higher hydrocarbons. Among the hydrocarbons, saturates and mono-
unsaturates (alkanes and alkenes) form the main components, with dienes and
acetylenic compounds (alkynes) normally present only as traces. As the gas is
subjected to different fractionations and reactions, so the relative proportions of
components obviously change.
Those hydrocarbon components which are not used in processing, or as fuel
gas, may be added to propane or butane product streams. These products may,
therefore, be totally saturated hydrocarbons, containing relatively small amounts
of each other, or mainly saturated, but with significant concentrations of unsatu-
rates.
1.3.1 Analytical requirements

Most gas streams measured only exist within the refinery boundaries, being
made, fractionated, reacted and combusted according to the process requirements
at any particular time. Analysis of these streams is aimed at optimizing the proc-
ess for which they are used, and it can be argued that consistency of measure-
ment between inlet and outlet streams is more important than absolute accuracy
of composition. (The alternative argument, that medium or long term consistency
can only be guaranteed by ensuring accuracy, is probably a better one.)
Product streams, such as propane, propene and butane, which are to be sold to
customers, require analysis for quality measurement, usually against a specifica-
tion. The specification, as for natural gas, will detail certain properties, but will
also require that there are not more than defined amounts of specific impurities.
Properties specified will be quite different from those for natural gas. This is
largely because the composition of the refinery product is dictated by the proc-
esses through which it is generated, which are controlled, whereas the composi-
tion of natural gas is subject to as little alteration from that found at the produc-
tion separator as the supplier can achieve. Although propane, for example, is
sold very widely for heating, the combustion characteristics of the product vary
very little between batches, whereas natural gas can show large differences. On
the other hand, LPG is sold and handled as a liquid, and so its vapour pressure is
an important property.
If an analytical method can be found that is capable of measuring the complex
mixture which is refinery gas, it should also be usable for simpler mixtures, such
as product streams. Although the mixture does not have as wide a boiling range
as natural gas, it contains extra components (hydrogen, carbon monoxide, un-
saturated hydrocarbons) which add their own complexity. It is, therefore, as with
natural gas, likely to need a complex analytical system, involving multiple col-
umns, or temperature programming, or both.
References p . 40
30 Chapter I
1.3.2 Analytical procedures

Separation of the inorganic gases, and of methane and the C, hydrocarbons
can be achieved by a molecular sieve/porous polymer combination such as is
shown in Fig. 1.16. Carbon monoxide elutes after methane on an activated mo-
lecular sieve, and the C, unsaturates can be separated from CO, and from each
other on an appropriate porous polymer. This is a well-established column com-
bination, and there is really no practicable alternative for the inorganic gases.
The temperature programmed porous polymer column shown in Fig. 1.15 would
be incapable of sensibly distinguishing 0,, N, and CO. Because of the presence
of CO, it is unlikely that the configuration would be exactly the same, or the or-
der of elution similar to that in Fig. 1.17; this is discussed further below.
'The hydrocarbons in Fig. 1.17 are separated according to boiling point, using
a capillary column. The boiling points of refinery gas hydrocarbons are given in
Table 1.1. In the table, component names are given in both IUPAC and the more
commonly used trivial form. The symbols are used to annotate the chroma-
tograms.
As Table 1.1 shows, there are significant boiling point differences, except for
the C, hydrocarbons (l-C4-, b.p. 4 4 3 C and i-C4-, b.p. -7.08"C). It is possible
to separate all components using a boiling point capillary column with a sub-
ambient temperature programme, but the method is not advisable where compo-
nent concentrations vary widely.
TABLE 1.1
REFINERY GAS NAMES, SYMBOLS AND BOILING-POINTS
Component Symbol Boiling point ("C)
Methane c1 -161.78
Ethane c2 -88.78
Ethene (ethylene) c,- -103.88
Ethyne (acetylene) c,= -84.93
Propane c3 -42.24
Propene (propylene) c3- -47.88
Propyne (Me-acetylene) c3= -23.62
Propadiene c3- -35.18
2-Me-propane (I-butane) i-C, -12.1 1
Butane (n-butane) n-C4 4.73
1-Butene 1-c4- -6.43
cis-2-Rutene c2-c4- 3.45
rmns-2-Butene t2-C4- 0.70
2-Me-propene (1-butene) i-C,- -7.08
1.3-Butadiene 1,3-C,-- -4.80
2-Me-butane (1-pentane) i-C5 27.59
Pentane (n-pentane) n-CS 35.83
There have been many references to columns and column combinations for
separation of all C, saturates and unsaturates [17-191. One of the earliest [20]
referred to the use of activated alumina, the surface of which was strongly ad-
sorptive to unsaturated hydrocarbons. When fully activated, retention times for
unsaturated hydrocarbons were excessive, and their peak shapes were poor, so a
degree of deactivation was recommended, using water or a combination of water
and silicone oil. This gave good separation, but was only consistent over a few
hours, as the water was stripped off by the dry carrier gas and the relative reten-
tions of saturated and unsaturated hydrocarbons changed significantly. It was
usually necessary to precede a days work with the injection of a relatively large
amount of liquid water, which would distribute itself over the alumina and allow
several hours of reasonably consistent separations.
Capillary columns packed with alumina were demonstrated [21] shortly after
the first reference, and later applied to refinery gas analysis [22]. In this applica-
tion, a humidification device was used for the carrier gas. This consisted of a
length of tubing packed with copper sulphate crystals, which, at ambient tem-
perature, assured a constant water content in the carrier gas, and hence a constant
polarity. In fact, with this device, there is the mechanism for fine-tuning the col-
umn behaviour. The vapour pressure of water above copper sulphate crystals is
constant at a constant temperature, and so the partial pressure of water in carrier
gas is controlled by adjustment of the total pressure in the humidifier. With the
arrival of porous layer open tubular (PLOT) capillary columns, alumina was
used in this form. The separations which had been incomplete with packed col-
umns were now baseline to baseline.
These columns were rapidly seen to give superior performance to any of the
alternatives, and became the state-of-the-art solution to the problem. The need to
maintain a constant level of moisture in the carrier gas was inconvenient, and so
alternative methods for deactivation were studied. The use of inorganic salts
proved to be effective, and potassium chloride treatment was the first such to be
commercially successful. KCI deactivated alumina, used with dry carrier gas,
gives excellent separation, but still has an affinity for moisture, which can alter
both absolute and relative retentions. When not in use, therefore, it is recom-
mended that the column should be kept at a relatively high temperature (e.g.
150OC) to avoid adventitious moisture. Columns are available with 0.32 mm in-
ternal diameter, with a 5 p m layer, or with 0.53 mm internal diameter, with a
10pm layer.
Figure 1.22 is a chromatogram of hydrocarbons on KCI-deactivated alumina.
The separations are good, such that most components could be measured even if
they were trace impurities. The separations which could be improved are those
of acetylene and propadiene from the C, saturates, and of 1-butene from trans-2-
butene. Further studies of alternative deactivating agents [23] showed that the
use of sodium sulphate rather than potassium chloride gave a phase with rather
References p . 40
32 Chapter I
greater polarity, but still good peak shapes. Figure 1.23 shows a typical chroma-
togram.
Comparison of Figs. 1.22 and 1.23 shows the effect on acetylene and propadi-
ene, which are eluted, well resolved in the latter case, between the saturated and
unsaturated C, hydrocarbons. The sodium sulphate deactivation also elutes
the C, unsaturates with more equal spacing than with potassium chloride treat-
ment.
Figure 1.24 shows the use of a column deactivated with KC1 for measurement
of impurities in propene. The product is about 25% propane and 75% propene,
with other hydrocarbon gases present at parts per million concentrations. The
propene peak is clearly overloaded, giving a tailing shape, but the trace compo-
nents are easily measured. Because the temperature programme for this separa-
tion is different from that in Fig. 1.22, the order of elution of n-butane and
acetylene can be seen to have changed.
As mentioned earlier, inorganic gases are separated on a combination of po-
rous polymer and molecular sieve columns. Although PLOT capillary columns
are also available with these stationary phases, the extra resolving power which
they offer is not necessary for routine analysis of light gases, and so they are
rarely used. The column combination used in Fig. 1.16, which produces the
separation of light components shown in Fig. 1.17, is obviously similar to what
is required here. The separation is more complicated, due to the presence of CO
n-C4
i44 1
. _.__
I I
0 min 10 min 20 min
Fig. 1.22. [Iydrocarbons on alumina/KCI. Column: 25 m X 0.32 mm i.d. PLOT alumina deacti-
vated with KC1. Temperature: 35-200C at 10"Cimin. Carrier gas: helium at 1.0 bar.
CH4 c2 C 2 '
"44
i44-
IL
1,344- -
II
0 rnin
I
10 rnin
I
1_
20 rnin
Fig. 1.23. Hydrocarbons on aluminaMa2SO4. Column: 25 m X 0.32 mm i.d. PLOT alumina deac-
tivated with Na$O,. Temperature: 35-200C at lO"C/min. Carrier gas: helium at 1.0 bar.
and unsaturated C, hydrocarbons in refinery gas, and so the direct elution of the
gases from molecular sieve before the CO, reaches the end of the porous poly-
mer column is less practicable.
I I
0 rnin 10 rnin 20 rnin
Fig. 1.24. Impurities in propene on alumina/KCI. Column: 25 m X 0.32 mm i.d. PLOT alumina
deactivated with KCI. Temperature: 100-200C at 5"C/min. Carrier gas: helium at 1.0 bar.
References p . 40
34 Chapter I
Figure 1.25 shows a layout which is sufficiently flexible. With valve V1 in the
inject position, as shown, carrier gas CG1 passes via the porous polymer and
molecular sieve columns to the detector. Column lengths are chosen so that all
the lightest components (02, N,, CH,, CO) are passed onto the molecular sieve
column before CO, elutes from the porous polymer. At this time, valve V2 is
switched to isolate ;he light components in the molecular sieve column, and CO,
and the C, hydrocarbons elute from the porous polymer column via restrictor A
to the detector. After they have been detected, valve V2 is returned to position 1
to allow the elution and measurement of the light components, and valve VI is
also switched to position 1, the sample load position. This allows CG1 to back-
flush C, and heavier components from the porous polymer column to vent, while
carrier gas CG2 continues the elution of light components from the molecular
sieve. The pressure of CG2 and the setting of restrictor A are adjusted so that the
carrier gas flow to the detector remains constant irrespective of valve positions.
Figure 1.26 is a typical chromatogram. Hydrogen, which is a component of
refinery gas, is not shown here. There are two reasons for this. Firstly, H, is
-
hardly retained at all on these columns, and is very difficult to trap on the mo-
lecular sieve with the other light components, and secondly because of the diffi-
culty associated with measurement of H, in He carrier gas due to the non-linear
relationship between thermal conductivity and H,/He composition. The solution
of using a mixed carrier gas was proposed by Purcell and Ettre [24] and dis-
cussed by Cowper and DeRose [25], but there are implications for the measure-
ment of other components, and it is not recommended for this application.
Hydrogen measurement can be carried out with a separate column/detector
system. using Ar or N, as carrier gas, or using He carrier, provided that the
Sample
Mol. sieve column
I
I
CG1 ! I I
Porous
polymer
1 1
-i column
Carrier Gas
t-- - -A Restrictor A
L
CG2
- 1 Vent
Fig. 1.25. Light gases analyser. Valve 1, position 2; valve 2, position I

c2 c2- Nq CH4
co
I I I I
0 min 5 min 10 min 15 min
Fig. 1.26. Light gases on porous polymer/molecular sieve. Column 1: Molecular sieve 5 A. Column
2: Combination of HayeSep Q and HayeSep T. Temperature: 50C. Carrier gas: helium at
25 ml/min.
sample size is kept sufficiently small so that non-linearity is not significant. With
a sufficiently well equipped chromatograph, all three separation systems can be
configured into one unit.
Figure 1.26 shows that this system measures C, and C, hydrocarbons, which
are also covered by the alumina PLOT column. Since the hydrocarbons represent
the bulk of most refinery gases, the approach would normally be to measure all
the hydrocarbons on the alumina, and to use the porous polymer/molecular sieve
combination just for the inorganic gases 0,, N,, CO and CO,. As with natural
gas analysis, C, or C, could be used as a bridge component, to allow for any drift
in relative detector response between calibration and analysis.
If the inorganic gases are present at trace levels, then a hydrogenator can be
used to convert CO and CO, to CH,, allowing sensitive detection by FID. With-
out invoking exotic and rarely used detectors such as helium ionization, in-
creased sample size is the only practicable solution for 0, and N,.
Figure 1.25 shows an arrangement for backflushing C, and heavier hydrocar-
bons while analysing the lighter components. The same need can arise for the
alumina column; C, and heavier components can be present in a refinery gas
sample, the result of which will be to require a substantially longer analysis time,
or to risk interference with subsequent analyses. If these heavier hydrocarbons
do not need to be analysed, then they can be backflushed to vent using a configu-
ration as shown in Fig. 1.27.
References p . 40
36 Chapter I
Whereas a valve was used for backflushing in Fig. 1.25, the configuration of
Fig. 1.27 uses pressure balancing [26]. The boiling point capillary column,
coated with a non-polar phase, separates the C, to C, hydrocarbons from any
heavier components. Pressure regulator PR2 is set to deliver a pressure which is
slightly higher than the natural pressure at the T-junction when carrier gas is
being delivered under the influence of PRl alone. The backflush switch, shown
in both the forward flow (upper) and backflush (lower) modes, either allows
carrier gas from PRl to flow through both columns in series, or interrupts this
flow, so that PR2 delivers continuing flow through the alumina column and re-
versed flow through the boiling point column. The T-junction between the col-
umns is made using a glass quick-seal connector into which the capillary column
is pushed so that the polyimide coating seals to the slightly tapered internal bore.
The appropriate time at which to operate the backflush switch is found by trial
and error, or, if a further T-junction i s fitted, by bleeding a small amount of the
non-polar column effluent via a restrictor to a second detector.
If there is a need to measure the heavier components, rather than just get rid
of them by backflushing, the configuration shown in Fig. 1.28 can be used. In
Fig. 1.27, the dimensions and material of the backflush switch are not very im-
portant, as it is not in the flow path encountered by measured components. When
the backflushed components are to be measured, a different configuration is re-
Sample Valve
Vent
Carrier Gas
Boiling Point
Capillary
P R2
Alumina
PLOT
Detector
Fig. 1.27. Rackflush of heavier components

LI
Sample Valve T
Boiling Point
Capillary
I
Heavier
PR2 Components
Capillary
Detector 1 Detector 2 cz]

Fig. 1.28. Measurement of heavier components.
quired. The backflush switch is replaced by a backflush valve, which must be of

the miniature-dead-volume type, compatible with capillary columns. When the
backflush valve is operated, PR2 supplies forward flow to the alumina column
and backflush flow to the boiling point column, diverting heavier components
into a third capillary column. This is chosen according to need, and can be a
further non-polar column, or a short alumina PLOT, for rapid elution of the
heavier components or groups. PR3 supplies a flow of carrier gas to the third
capillary column during forward flow to the others.
1.3.3 Sample handling

The range of samples varies from pure product to cracker streams after dif-
ferent stages of reaction, and also between those normally handled as liquids and
those which are always gases. LPG samples are normally handled entirely in the
liquid phase, and can be for the purposes of analysis. Chromatographic valves
are available which will easily cope with the pressures involved, and are capable
of injecting sample sizes of microlitres or smaller.
Since many of the streams to be analysed can only be handled as gases, it is
better to standardize on treating a11 samples in the gas phase, rather than to han-
dle some as gases and some as liquids. LPG product streams should therefore, be
fully vaporized before analysis. Sampling of LPG into a cylinder is straightfor-
ward, but an ullage space must be provided before the cylinder is transported or
Referencesp . 40
38 Chapter I
handled any further. This can be done by venting a proportion of the cylinder
contents, so that the cylinder contains some gas as well as the original liquid.
The different components in the sample will be distributed in different propor-
tions between the gas and liquid phases, but provided that the ullage space is
small, the liquid phase, containing the bulk of the sample, will still represent the
original composition. Where available, a constant pressure cylinder, which con-
tains a sliding piston seal between the two ends of the cylinder, should be used.
The sample side of the cylinder can then be filled with 100% liquid phase, and
the ballast gas, on the other side of the piston, provides the safety buffer.
Most analyses require injection onto more than one column system, even if
fitted into a single chromatograph, and it is therefore crucial that the sample is
uniform between the injections. Sufficient LPG should be totally vaporized to
form a gas sample large enough and with enough pressure to purge and fill all
the sample lines and injection devices. Depending upon the composition of the
sample, it may be necessary to heat the container into which the liquid is vapor-
ized, and to heat the lines through which the gas sample is conveyed to the injec-
tors.
1.4 CONCLUSIONS
From the foregoing, it can be seen that all the gas mixtures normally encoun-
tered in the petroleum industry can be separated into their individual compo-
nents. Having thus separated components, the usual requirement is to measure
them. This quantitative aspect has only been briefly mentioned, but is, of course,
of great importance.
Gas chromatography is not an absolute technique, in that the detectors de-
scribed above do not give a predictable response to any intrinsic property of the
separated components. Quantitative information arises from comparison, the in-
strument being calibrated with one or more mixtures of known composition, then
using the resulting response factors to convert the raw data (usually peak area,
occasionally peak height) to concentration. The quality of the calculated concen-
trations is fundamentally dependent upon how well the composition of the cali-
bration gas is known. Precision of analysis depends upon instrument character-
istics; accuracy depends principally on the quality of the calibration gas.
Detailed discussion of calibration gas preparation and certification is beyond
the scope of this chapter, but a few pointers are given below. Calibration mix-
tures can be prepared by gravimetric, volumetric or manometric techniques.
Most mixtures containing percentage concentrations can be prepared directly,
whereas lower concentrations may require one or more dilution stages to be
used. Some more reactive or adsorptive components, particularly at low concen-
trations (H,S being typical), require special precautions.
Gravimetric. Calibration gases of the highest quality can be prepared by this

technique. Pure gases are added to a cylinder, which is weighed before and after
each addition. Although the mass of the cylinder is typically orders of magnitude
greater than the masses of the added components, the accuracy achievable, with
suitable equipment and procedures, is excellent. The advantages include the fact
that masses translate unequivocally to molar amounts, and the technique pro-
duces a substantial quantity of calibration gas, under pressure in a cylinder. For
adsorptive components, cylinders which have been specially treated internally
can be used.
Volumetric. Volumetric techniques operate at or very close to atmospheric
pressure, and can be static or dynamic. For ideal gases, volumes at a constant
pressure are equivalent to molar amounts, and the corrections for real gases at
atmospheric pressure are small. Glass double-ended bulbs and gas-tight syringes
can be calibrated for volume from the mass of their water content. Total dis-
placement of the pure gas contents of a smaller container into a larger one, and
subsequent dilution with a complement gas gives good quality mixtures. Dy-
namic techniques are often preferred for minor and/or adsorptive components.
Various options exist, but for hydrocarbon gases, continuous injection from a
motor-driven syringe into a flowing diluent stream is usual. The uncertainties on
the composition are greater than with static methods, as measurement of flow is
less well defined than measurement of volumes, but the equipment will become
conditioned to adsorptive components.
Manometric. As with gravimetric mixtures, components are added succes-
sively to a cylinder, but the composition is calculated from pressure, rather than
mass increments. This technique allows the largest quantities of calibration gases
to be prepared, as the size of cylinder is not limited by the size of the balance, as
in gravimetric use. The accuracy is, however, relatively poor, since non-ideal
behaviour is much more pronounced at high pressure. Compression (2)factors
can be calculated for hydrocarbon gas mixtures with good accuracy, but these
calculations assume that the mixtures are homogeneous. The conditions under
which a cylinder is filled with different components do not encourage rapid
mixing within the cylinder. The calculated factors can therefore only be ap-
proximations.
1.5 ACKNOWLEDGEMENTS
I wish to thank British Gas plc, for permission to prepare and to publish this
chapter. I would also like to thank my many colleagues who assisted in its prepa-
ration, and Mr. A. Allott and Mr. R. Jackson, of Lindsay Oil Refinery Ltd., for
their valuable discussions on refinery gas analysis.
References p . 40
40 Chapter I
1.6 REFERENCES
I A. Melvin, Natural Gas:Basic Science and Technology, IOP PublishingiBritish Gas plc
(1 988).
2 G.J. van Rossum, ed., Gas Quality - Specification and Measurement of Physical and Chemical
Properties of Natural Gas,Elsevier, Amsterdam (1986).
3 IP 337178. Analysis of Non-Associated Natural Gas by Gas Chromatography, Institute of Pe-
troleum. London.
4 .4STM D1945 - 1981, Analysis of Natural Gas by Gas Chromatography, American Society
for Testing and Materials.
5 J.S. Stufkens and H.J. Bogaard, Anal. Chern., 47 (1975) 383.
6 1SO 6974 - 1984. Natural gas - Determination of hydrogen, inert gases and hydrocarbons up
to Cg - Gas chromatographic method, International Organisation for Standardisation.
- 1,. Huber and 14. Obbens, J. Chromatogr., 279 (1983) 167.
8 Varian Ltd., Application note No. 31.
9 IJ.S. Patent Application 061583,469,
I0 I.B. Angcll. J.H. Jerman, S.C Terry and S. Saadat, A Prototype Gas Analysis System using a
Miniature Gas Chromatograph, U.S. Department of Health and Human Services (1981).
11 J.B. Angell, S.C. Terry and P.W. Barth. Sci. Am., April (1 983) 36.
12 A. van Es: C. Cramers and J. Rijks, J. High Res. Chromatogr., 12 (1989) 303.
13 Chrompack I,td., Application brochure 501660.
11 R. Kenter, M. Struis and A.L.C. Smit, Process Control Qual., 1 (1991) 127.
I5 IS0 Ills 10723, Natural gas - Performance evaluation of analysers, International Organi-
sation for Standardisation.
16 E.H. Osjord and D. Malthe-Soerenssen, J. Chromatogr., 279 (1983) 219.
17 !I. DiCorcia and R. Samperi. J. Chromatogr., 107 (1975) 99.
18 N.C. Saha, S.K. Jain and P.K. Dua, J. Chromatogr. Sci., 16 (1978) 323.
19 D.R. Deans and 1. Scott, Anal. Chem., 45 (1973) 1137.
20 C.G. Scott, J. Inst. Petrol., 45 (1959) 118.
21 1. lialasz and E. Heine, Nature, 194 (1962) 971.
22 N.G. McTaggart, C.A. Miller and B. Pearce, J. Inst. Petrol., 54 (1968) 265.
23 N. Vonk, .I.dc Zeeuw, M. Mohnke and J. Buyten, 14th Int. Symp. on Capillary Chroma-
tography, Baltimore, MD (1992).
24 .I.J<. Purcell and L.S. Ettre, J. Gas Chromatogr., 3 (1965) 69.
25 C.J. Cowpcr and A.J. DeRose, The Analysis of Gases by Chromatography, Pergamon Press,
Oxford (1983).
26 D.R. Deans, J. Chromatogr., 18 (1965) 477.
CHAPTER 2
Advances in simulated distillation
D.J. Abbott
Esso Research Centre, Analytical Group, Abingdon, Oxfordshire OX13 6AE, UK
2.1 INTRODUCTION
Distillation is of major importance in the oil and petrochemical industries,

being by far the most widely used separation process. Many refinery units are
controlled by using distillation data and many products are sold to specifications
incorporating distillation data. There are a number of laboratory distillation tests
which are routinely used to determine the boiling ranges of crude oils and their
products. Crude oils are characterized by ASTM D2892 [l], which is a true
boiling point method involving a lengthy separation in a high efficiency still.
The method uses vacuum distillation to recover material with high boiling
points. A simpler method is ASTM D86, a single plate distillation applicable to
gasolines, middle distillates and similar products. There is also a low efficiency
vacuum distillation method, ASTM D1160.
Simulating distillation by gas chromatography was first reported by Eggerston
et al. in 1960 [2]. It is based on the fact that hydrocarbons are eluted from a non-
polar column in boiling point order, and the column is temperature programmed
until all the sample is eluted. Integration is done in fixed time slices, rather than
on individual peaks. The time axis is converted to temperature by running a
standard under identical conditions to that of the sample. This standard usually
consists of a range of normal paraffins whose boiling points are known accu-
rately, but it can be a mixture of aromatic compounds if highly aromatic samples
such as coal liquids are being analysed. Using data from the runs on the sample
and the standard, it is possible to calculate the percentage recovered at any tem-
perature, or vice versa. The calculation is best done by a computer, which can
also be used to measure the area slices and retention times required. Indeed, the
technique is ideally suited to automatic running, using a computer to control an
References pp. 52-53

42 Chapter 2
autosampler, generate data and print out a report. The computer can also use es-
tablished correlations to calculate various important parameters such as car-
starting index and vapour-lock index from the GCD data.
The development of GC distillation up to 1978 was very well reviewed by
Butler [3]. Since that time, essentially four methods for simulated distillation of
crude oils and petroleum fractions have become available, and these are now
considered in turn.
2.2 MIDDLE DISTILLATES AM) LUBE OILS
Simulated distillation of these fractions is covered by ASTM D2887, the

scope of which requires that their boiling ranges must be greater than 55"C, and
their final boiling points must be less than 538C as measured by the method.
This is the simplest of the various GCD methods because it assumes 100% re-
covery of the sample and response factors are not used. Either flame ionization
or thermal conductivity detectors are allowed, and a wide variety of stationary
phases and column lengths may be used. The method relies for control only on a
test for column resolution determined on a 1% solution of hexadecane and octa-
decane in n-octane. Unlike the gasoline GCD method, ASTM D3710, there are
no tests for sensitivity, noise, drift, skewing of peaks, or polarity.
2.2.1 Precision
The relative simplicity of the method does not mean that it is easy to obtain
results with a high level of precision, which is what the recipients of the data
usually want. The precision data in ASTM D2887, given in Table 2.1, has been
widely disbelieved for many years as too optimistic, and in fact the latest version
( 1989) notes that the data were not obtained in accordance with ASTM practices.
TABLE 2.1
ASTM D2887 PRECISION
O 6 Recovered Repeatability Reproducibility

("C, ("C)
~~
IBP 8
5 3
10-40 4
40-90 45
95 5.5
FBP 13.5
Advances in simulated distillation 43
Most laboratories practising GC distillation have found it difficult to achieve

these levels of precision, especially the reproducibility figures. This is probably
a reason for GCD not replacing physical distillation methods such as D2892 and
D86 in product specifications.
The most difficult points at which to achieve high precision are of course the
0-5% and 95-100% points. Sadly for the analyst, these are often the points of
most interest to customers. For example, the 95-100% points of a middle distil-
late can often be correlated with wax appearance temperature, and 0-5% of lube
oils can be correlated with oil consumption in engines. In consequence, refinery
pipestill operators monitor these points closely. Large sums of money may be
involved in achieving the optimum cut-points.
In view of the importance of precision, it is odd that there has been little pub-
lished work on ways of improving it. In the early 1980s, an Institute of Petro-
leum sub-panel [4] tried closely specifjring column, flow rates and temperature
programs, but failed to significantly improve precision. In 1983, Abbott [5] re-
ported that the poor precision was due, at least in part, to the lack of a common
method for calculating results from the raw data. Schwartz et al. [6] , in a paper
comparing simulated distillation by GC with SFC, made the same observation.
ASTM D2887 gives little guidance on the calculation, which is far from straight-
forward. It suggests that area summation be started at the beginning of the run
(or after a solvent exclusion gate), and consequently a lot of noise can be accu-
mulated if the sample does not start to elute quickly. There is no algorithm to
calculate the start of the sample. It suggests that the end of the sample be taken
as the point where the signal has reached a constant low value no greater than
0.01% of the total area counts. However, this will be dependent on the slice
width, which is not specified in the method, and therefore different computer
programs may choose the end points in different places.
A second major problem is baseline drift. D2887 states that subtraction of a
baseline blank from sample runs is probably more accurate than the traditional
approach of using matched columns and detectors. However, Abbott [5] showed
that baselines are not constant from one run to another, nor are they related to
each other by any simple transformation. Selucky et al. [7], in a paper on simu-
lated distillation of coal liquids, proposed a novel baseline correction procedure
which used baseline information from the actual experimental run before and
after sample elution. They showed that this approach gave better results than the
baseline subtraction method.
Abbott suggested a calculation procedure which addressed the problems of
sample detection and baseline drift simultaneously. Baseline drift could be com-
pensated for by either matched columns or by blank subtraction, but a limit was
put on the amount allowable. The procedure also made the use of a certified ref-
erence sample mandatory. This was to be used to determine a sensitivity factor
Referencespp. 52-53
44 Chapter 2
for the GCkomputer system, which was then used for subsequent samples. A
small scale cooperative study showed that the procedure greatly improved repro-
ducibility, especially the 0-5% and 95-100% points.
Most analysts performing simulated distillations naturally use commercially
available programs to do the calculations, and the authors of these programs
have interpreted the existing method in various ways. Fortunately, at the time of
writing (1992), an ASTM task force is developing a calculation procedure for
inclusion in a future version of D2887.
2.2.2 Capillary columns
ASTM D2887 allows the use of any column which meets its resolution crite-
rion and which elutes typical hydrocarbons in boiling point order. However, the
examples it gives are all with packed columns. Over the last 10 years or so the
use of capillary columns with internal diameters of circa 0.5 mm (wide-bore or
mega-bore) has become widespread [8-171. These columns can be used with
conventional injection systems and flow controllers, and have the advantage that
they usually produce little baseline drift.
This facilitates single column operation with baseline subtraction, A major
advantage of these columns is that it is often possible to elute much higher boil-
ing compounds than with packed columns. Of course, this means that they go
beyond the scope of D2887, which is limited to a final boiling point of 538OC,
and they will be discussed in more detail in Section 4.
2.2.3 Aromatics and heteroatoms
It is well known that not all hydrocarbons elute from non-polar columns in
boiling point order, and that compounds incorporating heteroatoms have differ-
ent detector response factors from hydrocarbons. D2887 addresses these prob-
lems in an appendix to the method. It shows that the deviation of simulated dis-
tillation boiling points from actual boiling points for 36 compounds, including
polycyclic aromatics and heterocyclics, does not introduce any significant error
when compared to physical distillation. A comparison of true boiling point data
(obtained on a high efficiency spinning band cohmn) and simulated distillation
data for a virgin gas oil, a high sulfur gas oil, and a highly aromatic gas oil
showed reasonable agreement. It is significant that the agreement was with
weight YOTBP data. In the early development of simulated distillation methods,
reviewed by Butler [3], there was disagreement about whether GCD correlated
with weight YOor volume % by TBP methods. The conclusion of the D2887 ap-
pendix was supported by Kiser and Malone [IS], working with coal liquids.
7'11t.y noted that none of the polycyclic aromatics mentioned in the ASTM
D2887 appendix had alkyl substituents, whereas real samples do. They under-
took a rigorous comparison of simulated distillation with direct TBP distillation
and also with TBP data computed from ASTM D86 data. They found good
agreement with weight % TBP data, and also noted that the repeatability of GCD
was better than that of the TBP data. Similarly, Pannell and Sood [19], also
working with coal liquids, found that there was a maximum 3% difference in the
cumulative weight % at any boiling point between GCD data and TBP data.
In another paper on coal liquids, Selucky [7] suggested that since the para-
ffinic scale and aromatic scale were the two extremes, a calibration curve
adjusted for aromaticity (e.g. from NMR measurement) would improve results.
Petroff et al. [20] reported that differences between calibration with n-paraffins
and aromatics could be avoided by the use of Dexsil 300 as stationary phase in-
stead of the more widely used non-polar silicone fluids. This was especially true
for a light cracking oil with 78% aromatics (Table 2.2), although OV 101 ap-
peared better at 0-5% recoveries.
However, the difference in the GCD results for Dexsil and OV 101 columns
was not very great for a naphtha which contained only 18% aromatics.
2.2.4 Multi-element speciation
An interesting application of simulated distillation was reported by Buteyn

and Kosman in 1990 [21], in which it was combined with elemental analysis.
Many refined products have to meet elemental specifications, e.g. sulfur in diesel
fuel, and by performing GC distillation with an atomic emission detector, the
authors demonstrated that an elemental analysis on each 1% fraction could be
TABLE 2.2
SIMULATED DISTILLATION OF AROMATIC SAMPLE
ov 101
n-A1kane Aromatic n-Alkane Aromatic
calibration calibration calibration calibration
~~ ~ ~
0 168 154 159 159

5 200 192 193 196
10 221 219 212 218
30 244 239 234 245
50 256 255 248 262
70 272 272 263 279
90 295 300 285 307
95 303 308 294 319
100 323 332 3 14 344
Referencespp. 52-53
46 Chapter 2
achieved. Further information on this is given in Chapter 7 on the atomic emis-

sion detector.
2.3 GASOLINES AND GASOLINE FRACTIONS
The ASTM method for gasolines, D37 10, is much more complex than that for
middle distillates and lube oils which has just been discussed. This is primarily
because individual hydrocarbons in the gasoline boiling range have quite differ-
ent response factors in either flame ionization or thermal conductivity detectors.
The scope of the method limits the final boiling point to 260C, and response
factors are determined from a standard such that the results are given in volume
YOwhichever detector is used. Conditions are selected such that iso-pentane and
lighter saturates are measured individually, and heavier compounds are measured
as pseudo-components of narrow boiling range. The calibration mixture for this
method is a complex one of pure compounds covering the boiling range. It is
used not only for the conversion of retention time to boiling point, but also for
response factors which are applied to individual compounds and to the pseudo-
components. It is also used for system performance checks such as resolution,
sensitivity, noise, drift and peak skewness.
The precision of the method depends on the shape of the boiling range distri-
bution curve. Both repeatability and reproducibility vary with the rate of change
of temperature with percent recovered. The precision data are given as Tables
2.3 and 2.4. It is worth noting that a significant problem with gasoline samples is
sample integrity. Gasolines contain high volumes of very low boiling compo-
nents which are easily evaporated if stringent precautions are not taken. This is a
major problem in cooperative testing to obtain reproducibility data, and it can be
TABLE 2.3
Repeatability ("C) at dTldV
0 1 2 3 4 6 7 S 11 17 22
1 1 1 1 1 1 1 1 1 1 1
I I 1 1 1 1 1 1 - - -
I 1 1 1 1 I I 1 - - -
I 1 1 1 1 1 1 1 - - -
1 2 2 3 4 6 - - - - -
I 2 2 3 4 6 S 10 - - -
- - 1 2 3 4 6 7 - - -
- - - - - - - 3 3 3 3
- - - - - - - - 3 3 3
TABLE 2.4
Reproducibilitya (C) at dT/dV
0 1 2 3 4 6 7 8 I1 17 22
IBP 4 4 4 4 4 4 4 4 5 6 7
1 3 3 3 3 3 3 3 3 - - -
5 3 3 3 3 3 3 3 3 - - -
10 3 3 3 3 3 3 3 - - - -
20 3 4 6 9 13 17 - - - - -
30-90 3 5 7 11 15 20 26 31 - - -
95 - - 6 9 13 17 21 26 - - -
99 - - - - - - - 11 13 18 20
FBP - - - - - - - 11 14 20
aFor thermal conductivitydetectors. For FID, reproducibility is the same except in the range 20-
95%, where R(FID) = 0.9R(TCD).
-, outside the range observed in the cooperative study.
a problem when obtaining repeatability data, or when using a quality control

sample within a single laboratory.
There has been little published work on the simulated distillation of gasolines
since the review by Butler [3]. Zuber and Bart1 [22] published an interesting pa-
per on quality control of aviation fuels using a modification of D3710. They
noted that GCD has not replaced the physical distillation method D86 in practice
because boiling range distribution is only one of a series of properties to be
checked within the quality control procedure for fuels. These properties include
vapour pressure and flash point, and to calculate these accurately, they required
more precise initial and final boiling points than those given by D3710. They
modified the method by using a 50-m capillary column with 1 p m film thickness,
thus eliminating the need for sub-ambient cooling. The increased resohtion of
the capillary greatly improved GCD data at the low and high boiling ends, and
thus improved the accuracy of calculated properties.
2.4 HEAVY LUBE OILS AND RESIDUES
Simulated distillation of these fractions has become important because their

boiling range distribution provides information on feedstocks for conversion
units such as hydrocrackers and visbreakers. Also ASTM D2887 does not cover
all lubricants, especially synthetic ones such as poly-alpha olefins. In conse-
quence, this has been the most active area of research in simulated distillation
during the last 10 years. Prior to that time these types of sample could only be

48 Chapter 2
subjected to GCD by using an internal standard procedure with either backflush

of the whole column or, preferably, a pre-column. However, a proposed ASTM
method using this approach was never advanced to method status because of a
lack of adequate precision.
Assigning boiling points to alkanes higher than C60 is a problem because they
cannot be distilled even under vacuum. They are obtained by extrapolation of the
curve correlating n-alkane boiling points with carbon number [23]. Some authors
emphasise the "unreal" nature of the values [24].However, the people who use
the data, such as pipestill operators and lubricants formulators, rarely question
the results since they are usually interested in comparative values.
In 1985. Luke and Ray [S] published their paper on GCD of atmospheric resi-
dues using short (6 m) pyrex capillary columns with only 0.15 p m film thickness
of the silicone stationary phase OV-1. They used a stainless steel capillary re-
strictor so that the hydrogen carrier gas could be flow controlled, and they used
cold on-column in-jection to avoid discrimination. The column was programmed
from 30 to 390C and it was shown that residues boiling up to 650C (carbon
number 70) could be analysed by this technique.
The paper by Luke and Ray was closely followed by one from Trestianu et al.
191, which described a dedicated system for GCD of heavy petroleum fractions.
The system utilized pressure and flow control for the carrier gas and an auto-
matic on-column injector. They demonstrated the elution of n-alkanes up to
Cl20 within the temperature program, which went to 43OoC, and up to C140
during the final isothermal hold. Repeatability of the dedicated system was
shown to be very good, and the lower part of the distillation curves fitted very
well with those obtained by D2887.
Other authors also reported GCD of heavy fractions using capillaries [ 10-1 71,
and Firor [ 151 showed that fused silica capillaries could be prepared for use up
to 400"C, thus obviating the need for aluminium-coated silica or glass columns.
Recently, metal capillary columns with the stationary phase chemically bonded
to the internal surface have been introduced [25].
There are alternatives to high temperature GC for the simulated distillation of
high boiling fractions, such as supercritical fluid chromatography (SFC) and
vacuum therinogravimetry (TGA). A concern with high temperature GCD de-
scribed earlier is that petroleum fractions are known to decompose at tempera-
tures above about 350C. Decomposition depends not only on temperature but
also on sample composition and residence time at high temperature. SFC has
enjoyed a resurgence of interest in recent years, and Schwartz et al. [26] reported
its first application to sitnulated distillation. They used a 10-m 5 0 p m internal
diameter capillary column coated with a non-polar stationary phase. The column
had an integral restrictor which gave 3.4 ml/min flow when used at 1800 psi of
liquid carbon dioxide and at room temperature. An HPLC valve was used as the
injection system, which was connected to the column by means of a simple

splitter. At an oven temperature of 100C and using a linear pressure program of
2000-5500 psi of carbon dioxide, they found that C80 eluted in around 21 min,
and that retention time repeatability of the n-paraffin calibration standard was
excellent. The method was applied to an atmospheric residuum with a boiling
range of 345-770C.
In a later paper, Schwartz et al. [6] gave a more detailed account of simulated
distillation by capillary SFC. They also compared the technique to GCD and to
simulated distillation by TGA. In fact, they used TGA with and without vacuum
to show that an Arabian heavy crude residue started to decompose thermally at
around 370C and reached a maximum at 430C. Decomposition products were
confirmed by using TGA linked on-line to a GUMS system. The capillary SFC
conditions which they studied included column type, fluid density and tempera-
ture, and discrimination effects in the inlet splitter. This last factor is important
because, while the sample may be soluble in a solvent prior to injection, precipi-
tation may occur just after injection when the sample is exposed to carbon diox-
ide at relatively low density. They concluded that an oven operating temperature
of 100C overcame this problem. The pressure program was optimized so that
samples with a boiling range of 120-760C could be analysed. Three lubricating
oils were then analysed by GCD (ASTM D2887) and by capillary SFC, and a
regression analysis indicated no systematic difference between the two sets of
results. The authors also compared SFC with a vacuum TGA method of simu-
lated distillation described in detail in the paper. Two sets of atmospheric resi-
dues were cut into five fractions using short path distillation at high vacuum, and
each fraction was analysed by both methods. Regression analysis of the data
showed that SFC gave higher values than VTGA at low boiling points while the
reverse was true at higher boiling points. The authors considered that this might
have been caused by the calibration method used for VTGA, which was non-
linear in the upper temperature region.
Schwartz published a further paper on simulated distillation by SFC in 1988
[27], this time using packed columns. The advantages of these were said to be
high loadability (i.e. no splitter), good quantitation, and faster analysis time. A
10-cm column packed with 7 p m polysiloxane PM allowed elution of ClOO in
about 20 min. A comparison of packed SFC, capillary SFC and VTGA showed
good agreement. An important advantage of packed SFC was its ability to accept
undiluted samples such as crudes, and this is discussed in Section 5.
As noted by Thomson and Rynaski [28], simulated distillation by SFC has not
been widely adopted as problems have been encountered with the injection of
high boiling samples. However, they show that much improved sample introduc-
tion techniques are now available, and in their analysis of wax samples they
found that SFC and high temperature GC gave equally good results.

50 Chapter 2
Other approaches to simulated distillation of heavy fractions have also been

reported. Petroff et al. [20], in their wide-ranging paper on the application of
GCD to petroleum product control, described the use of temperature pro-
grammed injection and the backflush of a pre-column. They used an internal
standard of C 1 4 4 1 6 to calculate sample recovery. DiSanzo et al. [29] used a
very similar procedure and reported excellent long-term precision for a lubricat-
ing oil with boiling range 390-590C. Abbott [30] described a valveless method
of backflushing which used flow controllers rather than the pressure controllers
used in a Deans system [31]. The advantage was that a wide temperature pro-
gram could be used, such as for heavy residues. Finally, Curvers and van den
Engel [24] described commercially available equipment incorporating a tempera-
ture programmed injector and a wide-bore high temperature column.
2.5 CRUDE OILS
Crude oils pose all the problems of the previous three sections for simulated
distillation: precision, variable response factors, non-recoverable residues, etc.,
and a viable method for crudes has been a goal for chromatographers since the
early days of the technique. As mentioned in the Introduction, TBP data for cru-
des can be obtained by ASTM D2892, but the method can take days to complete.
TBP data on crudes is very valuable to logistics planners and pipestill operators,
so a simulated distillation method taking only 2 h would be an extremely attrac-
tive proposition.
The ASTM published a proposed method in 1976, which involved analysing
the sample with and without the addition of an internal standard of C14 - C17 n-
paraffins. This allowed calculation of sample recovery, typically up to 538C.
The method was never advanced to standard status because of inadequate preci-
sion obtained in cooperative testing programmes. Even so, the method is used,
and Maynard and Michalik [32] report that the use of robotics to prepare the
samples has greatly improved precision. Analysis of a Mexican crude with boil-
ing range 200-450"C and 22% residue gave a standard deviation of 1.1% for the
residue and 2.3"C throughout the boiling range.
Petroff et al. [20] used a more sophisticated version of the proposed ASTM
method. Their equipment incorporated a programmable temperature vaporizer
and a pre-column which was backflushed to prevent heavy residues from con-
taminating the analytical column. They found good agreement between GCD and
D2892 results for Kuwait and Boscan crudes.
Schwartz [27] used packed column SFC for simulated distillation of Maljamar
crude. In order to minimize the sample volume placed on the column, a fast
valve injection time was used. He was able to show the separation of early
eluting sample components, and presumably the technique was able to go up to

C 100 as before (Section 4), but unfortunately the paper did not include precision
data.
Coulombe and Duquette, in a paper on SFC of crude fractions [33], noted that
response factors for sulfur and nitrogen containing compounds can be quite dif-
ferent from hydrocarbons. This appears to be an area which has received insuf-
ficient attention, since crude oils vary greatly in their content of heteroatoms.
2.6 PROCESS CONTROL AND OTHER APPLICATIONS
Simulated distillation is probably used to control processes in hundreds of lo-

cations around the world, but mostly off-line. However, as long ago as 1975 Bird
and Kimball described the application of on-line gasoline GCD to refinery proc-
ess control and blending operations. A significant advance since the review by
Butler [3)]has been the introduction of simulated distillation by on-line SFC,
reported by Levy [34]. As described in Section 4,SFC extends the boiling range
which can be analysed to as high as 800C, and thus simulated distillation data
can now be obtained on-line for refinery units processing heavy residues. The
system incorporates a supercritical fluid extraction cell, which is necessary be-
cause of the viscous nature of the samples. It also means that the sample is in-
jected as a dilute solution in carbon dioxide, thus avoiding many of the injection
problems found in SFC, and it ensures that the capillary column is not over-
loaded.
In a related application, Abbott reported that GCD data could be used for the
automatic identification of leakages in refineries [35]. Traditionally, unknown
samples in refinery sewer systems have been identified by comparing them with
a library of fingerprints, preferably obtained by capillary GC. This approach
uses only the areas or heights of easily recognised peaks, and it requires an ex-
perienced operator to interpret the results. GCD data is in effect a way of digitiz-
ing a chromatographic trace, and Abbott combined it with peak area data to pro-
duce a system in which unknown samples were identified by a computer pro-
gram. Obviously many of the leakage samples are weathered, but this problem is
solved by including data from artificially weathered products in the computer
database.
GC distillation can be used to determine gasoline fuel dilution of used engine
oils, although there is a separate ASTM method (D3525) for this purpose. How-
ever, GCD cannot be used for diesel fuel dilution of engine oils because of the
substantial overlap of fuel and lube. Fuels have a typical boiling range of C8-
C26 and lubes from C18440. Adlard and Davies overcame this problem by us-
ing a GCD type of analysis, in that the chromatogram was acquired as area time

52 Chapter 2
slices rather than peak areas [36].A computer program then compared data from
the fresh oil with that from the used oil, and from the difference in areas the per-
centage of fuel in the lube was calculated. The computer program was very so-
phisticated, being able to compensate for differences in retention time and in
sample size.
Cartellieri and Tritthart used GCD to analyse the oil soluble fraction of par-
ticulates emitted by diesel engines [37] so that they could estimate the propor-
tions attributable to the fuel and lube. They did this by constructing a calibration
graph of the 50% points of a range of fuel/lube mixtures against composition.
The 50% points for samples were determined and the proportion of fuel and lube
obtained from the graph. It was necessary to determine a calibration graph for
each fuel and lube being used (nine standards), but by using an autosampler the
authors were able to analyse a large number of samples.
2.7. CONCLUSIONS
It is now over 30 years since the first report of GC distillation [2]. The tech-
nique has been very successful in that it is now in widespread use and is used for
quality control in hundreds of locations. It is usually the method of choice for
unknown samples such as spillages, since it can be run quickly and cheaply.
However, the technique has not replaced older methods of physical distillation in
specifications, and in that respect has not attained its full potential. The major
reason for this is the lack of good reproducibility data discussed earlier, and it is
to be hoped that this problem will be solved. Another major challenge is the de-
velopment of a viable method for crudes, which would have large economic
benefits in many laboratories. The introduction of SFC has extended the scope of
GCD, and provided that good, easy to use instrumentation becomes available, it
should have a bright future.
2.8 REFERENCES
t ASTM methods can be found in the current edition of ASTM standards.

2 F.T. Eggerston, S.Groenings and J.J. Holst, Anal. Chem., 32 (1960) 904.
3 K.D. Butlcr. in K.11. Altgelt and T.H. Gouw (Eds), Chromatography in Petroleum Analysis,
Marcel Dekker. New York (1979).
Institute of Petroleum sub-panel ST-G-6B, unpublished work.
D.J. Abbott, J. Chromatogr. Sci., 21 (1983) 425.
I-I.E. Schwartz, R.G. Brownlee, M. Boduszynski and F. Su, Anal. Chem., 59 (1987) 1393.
1.G. Southern, A. Iachelli, D. Cuthiell and M.L. Selucky, Anal. Chern., 57 (1985) 303.
I.. Luke and J.E. Ray, J. High Res. Chromatogr., 8 (1985) 193.
S. Trestianu, G. Zilioli, A. Sironi, C. Saravalle, F .Munari, M. Galli, G. Gaspar, J.M. Colin
and J.L. Jovelin, J. High Res. Chromatogr., 8 (1985) 771.
10 A.D. Bashall, Am. Lab., 19 (1987) 56.

11 A. Rastogi, J. High Res. Chromatogr., 10 (1987) 479.
12 N.R. Warren and B.M. Lawrence, Lab. Pract., 36 (1987) 80.
13 F. Noel, J. High Res. Chromatogr., 11 (1988) 837.
14 R.L. Firor, Am. Lab. 21(2A) (1989) 32.
15 R.L. Firor and R.J. Philips, J. High Res. Chromatogr., 12, (1989) 181.
16 P.S. Spock, M.V. Robillard and R.E. Long, Adv. Inst. Cont., 45 (1990) 593.
17 M. Dorbon, S. Lamaison and A. Chevalier, J. Chromatogr., 557 (1991) 155.
18 M.D. Kiser and D.P. Malone, Am. Chem. SOC.Div. Fuel Chem., 27 (1982) 114.
19 R.B. Pannell and A. Sood, J. Chromatogr. Sci., 20 (1982) 433.
20 N. Petroff, A. Hoscheitt and J.P. Durand, J. Chromatogr., 395 (1987) 241.
21 J.L. Buteyn and J.J. Kosman, J. Chromatogr. Sci., 28 (1990) 19.
22 K. Zuber and P. Bartl, Fuel, 68 (1989) 659.
23 0. Glinzer, Erdoel und Kohle, 38 (1985) 213.
24 J. Curvers and P. van den Engel, J. High Res. Chromatogr., 12 (1989) 16.
25 Chrompack, Middelburg, The Netherlands.
26 H.E. Schwartz, J.W. Higgins and R.G. Brownlee, LC-GC 4 (1986) 639.
27 H.E. Schwartz, J. Chromatogr. Sci., 26 (1988) 275.
28 J.S. Thomson and A.F. Rynaski, J. High Res. Chromatogr., 15 (1992) 227.
29 F.P. DiSanzo, C.J .Lowther, R.E. Yoder and J.L. Lane, J. Chromatogr. Sci., 26 (1988) 39.
30 D.J. Abbott, J. High Res. Chromatogr., 7 (1984) 577.
31 D.R. Deans, Chromatographia 1 (1968) 18.
32 J.B. Maynard and W.A. Michalik, J. Chromatogr. Sci., 26 (1988) 290.
33 S. Coulombe and G. Duquette, Am. Chem. SOC. Div. Fuel Chem., 33 (1988) 920.
34 G.B. Levy,Am. Lab., (1986) 12.
35 D.J. Abbott, J. High Res. Chromatogr., 9 (1986) 598.
36 E.R. Adlard and R.E. Davies, Petroanalysis 81, Wiley, New York (1982).
37 W. Cartellieri and P. Tritthart, SAE technical paper 840418 (1984).
E.R. Adlard (Ed.), Chromatographyin the Petroleum Industry
CHAPTER 3
The chromatographic analysis of refined

and synthetic waxes
Arthur Barker
Dussek Campbell Ltd, Thames Road, Crayford, Dartjord, Kent DAl4QJ, UK
3.1 INTRODUCTION
During the petroleum refining process, various residues containing wax are
produced: tank bottoms, lube stock residues and distillate residues. These oily
wax materials, or slackwaxes, can have the oil removed to produce microcrys-
talline, intermediate and paraffin waxes. They are complex materials containing
straight chain and branched chain alkanes, cycloalkanes and very small amounts
of aromatics. The ratio of straight chain to branched chain alkanes varies accord-
ing to the type of wax. Microcrystalline waxes have the highest proportion of
branched chain alkanes (see Table 3.1). Also the carbon number distribution of
waxes varies according to the crude oil used and the degree of refining.
Materials with wax type properties can be synthesized from several routes.
The Fischer-Tropsch process is used to produce a unique series of high melting
waxes containing mainly straight chain alkanes. Polyethylene wax materials,
produced from the polymerization of ethylene, have a higher molecular weight
distribution than most petroleum waxes and consist almost wholly of even-
numbered straight chain alkanes. Originally polyethylene waxes were obtained
as a by-product in the manufacture of high density polyethylene. However, over
the past 5 years there has been a rapid growth in specialized processes to pro-
duce polyethylenes of narrow molecular weight distribution. There are also new
processes to manufacture synthetic low molecular weight waxes.
Waxes are usually sold according to average physical properties (such as
melting point, viscosity, or mean molecular weight), as the actuaI properties of
each batch will vary depending on the refinery process (typical values are shown

56 Chapter 3
TABLE 3.1
PERCENTAGE STRAIGHT, BRANCHED AND CYCLO ALKANES IN TYPICAL PETRO-
LEUM WAXES
Type of alkane Type of way
Paraffin wax Intermediate wax Microcrystalline wax
% Straight chain alkanes 65-95 -60 20-40

% Branched chain alkanes 3-20 -15 15-40
% Cyclo-alkanes 3-1 5 -25 -3 5
Carbon number range 1840 20-60 20-80
in Table 3.2). This is particularly true of petroleum waxes which also vary ac-
cording to the crude oil source. Most refined and synthetic waxes are not nor-
mally used in industry as single substances, but are blended together, with pos-
sibly other materials added. This gives the specific physical and chemical prop-
erties for the end use. Wax blends are important as adhesives and coatings in
packaging, as additives to inks and rubber components and in many other prod-
uct areas [ 13.
Refined and synthetic waxes are complex multicomponent materials of great
variety. Their structural properties, i.e. molecular weight distribution (or carbon
number distribution) and straighthranched chain nature, have a direct bearing on
physical properties such as melting point, crystallinity, hardness, adhesion,
flexibility and viscosity [2-51. Since the early 1980s there has been an expansion
in demand for more complete analysis of these materials (and blends of them)
for both quality control and product development. This has included gas liquid
Tr\BI,E 3 2
TYPICAL PHYSICAL PROPERTIES OF WAXES AND POLYWAXES
Type of material Typical physical properties
Melting point Viscosity Penetration

(in "C) (in C P S ) ~ (dmm125"C)
Paraffin wax 4646 2.0-3.4 9-20

Intermediate wax 6674 3 3-7.9 -20
Microc 60-93 7.9-21 2-3 0
Sasol c1 -100 8 2
P o l y ~ a x500 86 38 7
Polywm 1000 113 74 1
Polywax 2000 125 270 0.5
~~ ~~ ~~ ~
"\'iscosity of petroleum waxes is measured at 9 9 T , Sasol at 120C and Polywaxes at 149C.

The chromatographic analysis of refined and synthetic waxes 57
chromatography (GLC) for carbon number distribution analysis and size exclu-
sion chromatography (SEC) for synthetic wax analysis. Wax additive separation
and food contact analysis can be carried out using high performance liquid
chromatography (HPLC). There is a growing interest in using supercritical fluid
chromatography (SFC) for wax analysis, but this type of chromatography has yet
to find a niche.
3.2 GAS LIQUID CHROMATOGRAPHY
3.2.1 Establishing present technology

The GLC analysis of waxes can separate each of the straight chain alkanes (n-
alkanes) from the branched and cyclo-alkanes (iso-alkanes) of the same molecu-
lar mass (see Fig. 3.1). Using the best possible capillary GLC technique, the al-
kanes can be separated up to approximately C80. At this point, the branched and
straight chain molecules tend to merge, so that all the alkanes of each carbon
number are detected up to approximately C 120 (approx. molecular weight of
1900). This precise form of separation of wax materials is known as the carbon
number distribution.
Waxes are mainly non-polar and thermally stable; therefore absorption onto
GLC columns is not a problem and relatively inert stationary phases can be used.
The complex and extensive carbon number distribution analysis of waxes pro-
duces a high demand on the separation efficiency and thermal stability of any
GLC column used, particularly when it is taken above 350C. Because of the
C30
C25 I
STRAIGHT
CHAIN ALKANES ---- >
c35
A 10
f
20
I
30 4ominrtes
Fig. 3.1. Packed column GLC analysis of a typical rubber wax blend. 1.P method 372/85
(modified), 150-375C at 5C m i d .

58 Chapter 3
wide boiling point range of wax components, temperature programming is used.

For each wax sample, the repeatability of alkane retention data depends on accu-
rate oven temperature regulation and carrier gas control. In quantitative analysis
the design of both the injector and detector is important, as well as the mode of
data acquisition and calculation.
The basic techniques for the GC separation of all petroleum waxes were es-
tablished in the 1960s [6,7]. In 1970, the stable stationary phase Dexil 300 was
introduced [ S ] , to be used up to 400C to separate natural waxes [9]. Also in
1970, the first short capillary column was used to rapidly separate a microcrys-
talline wax up to carbon number C58 [lo]. In the 1970s, the basic principles of
capillary GLC and bonded stationary phases were established. However, al-
though microprocessor controlled data stations were used for data handling, GC
instrumentation was still based on transistors and resistors. The ASTM put for-
ward a proposed general method for the packed column GLC analysis of waxes
[ 1 11, but temporarily abandoned it for standardization. It was left to the IP Wax
Panel ST-G-GA to carry out round robin testing in 1979 and produce a packed
column method for carbon number distribution of paraffin wax by gas chroma-
tography [ 121.
The work carried out by the IP Wax Panel optimized the conditions for the
quantitative analysis of paraffin waxes up to 350C maximum separation tem-
perature. Dual columns containing an inert packing coated with a silicone gum
rubber stationary phase were used. However, the method did not address the
need for accurate quantitative analysis of microcrystalline waxes or wax blends,
which was then becoming important for quality control in the rubber industry.
There are several problems associated with the packed column GLC methods of
analysis of waxes [ 131:
1. From the production of waxes in the refinery to the end-user, different
GLC methods were used for carbon number distribution analysis (with
different column sizes, packing and temperature programs). This led to
different degrees of separation of branched and straight chain alkanes.
2. The flame ionization detector (FID) for a packed column GLC does not
have a linear response factor for alkanes above C32. Unless the instru-
ment is calibrated for response then inaccurate quantitative results are
achieved.
3. Packed column analysis of waxes is inaccurate because the straight and
branched chain alkanes cannot be resolved sufficiently to measure the
peak areas with any certainty.
4. Different methods of data handling, including peak area measurement and
carbon number distribution calculation, are used in different laboratories.
Harmonization between laboratories analysing wax blends and microcrys-
talline waxes is very difficult.
A typical packed column chromatogram of a rubber wax blend is shown in

Fig. 3.1. Although most petroleum GC analysis has now changed to using capil-
lary columns, it is only recently that the wax industry has abandoned packed
column methods for capillary column analysis.
3.2.2 The 1980s revolution

Many research ideas that were initiated in the late 1970s became commer-
cially practical in the early 1980s. The use of high temperature gas liquid chro-
matography (HTGLC) for the accurate analysis of waxes would not be possible
without the fused silica column containing a bonded phase, the modern micro-
processor controlled GC and the rapid growth in sophisticated data handling
techniques.
Glass capillary columns used in the 1970s were expensive to make, very frag-
ile and had an active inner surface that did not readily take stationary phases for
HTGLC .The introduction of the fused silica capillary column [14] with its inert
inner surface (providing a stable base for bonded phases) and outer polyimide
coating (giving the column strength and flexibility), made the detailed separation
of waxes possible. This coincided with the development of stable bonded phases
capable of being used to 350C without excessive bleeding from the column
[15,16]. Although before 1980 packed column GLC had been used to separate
waxes up to an oven temperature of 400C, it was not until 1985 that fused silica
columns were reported to be used in this manner. However, all the initial devel-
opments in capillary HTGLC were made in simulated distillation research for
petroleum refining. This technique is not required to give the resolution neces-
sary for the detailed separation of waxes. It was not until the late 1980s that pa-
pers on the high resolution, HTGLC separation of waxes were published.
The GC equipment of the 1970s, was inadequate for the high resolution sepa-
ration of waxes. The early 1980s saw a rapid development in GC design. Ovens
were redesigned with thin metal walls to reduce thermal lag and allow accurate,
reproducible temperature programming. Accurate diffusion-free pressure regula-
tors and low flow rate, mass flow controllers became available allowing accurate
control of the carrier gas at high oven temperatures. Microprocessor controlled
equipment enabled more precise and flexible regulation of the column oven and
detector temperatures to be obtained. The commercialization of on-column and
programmed temperature vaporizer (PTV) injectors revolutionized sample intro-
duction, minimized sample discrimination and allowing ,accurate quantitative
analysis above C30.
Microprocessors also revolutionized data handling and quantitative analysis.
The demands for more detailed analysis (including further calculations such as
carbon number distribution), data storage and comparison of chromatograms

60 Chapter 3
have now led to the common use of PC technology and specialized sofhvare for
wax analysis.
3.2.2.I Sample introduction
Rcfore a sample can be introduced into a capillary system it has to be dis-
solved in a suitable high purity solvent. The high molecular weight alkanes (i.e.
above C40) in microcrystalline and synthetic waxes tend to be insoluble in most
common solvents at room temperature. When an injector is used in the split
mode then the sample concentration has to be high (at least I%), and the solvent
and syringe needle have to be warm to ensure that the higher molecular weight
components do not condense. This means that a rapid, skilful injection tech-
nique, which is difficult to automate, has to be used to produce accurate results.
With on-column injection (including splitless PTV injection), a very dilute
sample of wax (1-3 mg ml-l) has to be used so as not to overload the thin-film
columns. For alkane standards, the concentration of each alkane is reduced to
0.1 mg ml-I. This still presents a problem as to what solvent can be used, particu-
larly if the method is to be automated.
Cyclohexane [ 171 and carbon disulphide [ 181, both on their own and as a 1:1
mixture, have been used as solvents for polywaxes. Mixtures of 1 :1 decane and
carbon disulphide, or xylenes and carbon disulphide, or iso-octane and carbon
disulphide [ 191 have been used for crude oil/wax mixtures and polywaxes. Both
carbon disulphide and xylenes are now considered as undesirable solvents for
normal use in a laboratory. Cyclohexane and heptane are practical solvents for
the analysis of refined waxes, but require heating to solubilize synthetic waxes.
The methods of sample introduction into capillary GLC systems have been
very ably covered in a book edited by Sandra [20]. Here, it is intended to em-
phasize and expand on the principles that affect HTGLC. The mode of injection
is of prime importance in the accurate quantitative analysis of complex, high
boiling point alkane mixtures. A limited amount (0.1-1 p l ) of sample must be
injected rapidly into a small volume so that it can transfer to the capillary col-
umn as a narrow band and be separated into its components without decomposi-
tion, peak broadening, or tailing [21]. The design of the injector is very impor-
tant in achieving an unbiased response for each component over the whole car-
bon number range.
The two most common injection techniques used in HTGLC are the on-
column injector invented by Schomburg and Husmann [22] and the Program-
mable Temperature Vaporizer (PTV) introduced by Vogt et al. [23]. There has
been continuous development of both these methods of injection over the past
I0 years.
On-column injectors are now designed so that a dilute solution of the sample
is injected through a septumless valve directly into a capillary column. The in-
The chromatographic analysis of rejined and synthetic waxes 61
jection point is externally cooled below the oven temperature to focus the sample
as a narrow band at the entrance of the column. This secondary cooling is
switched off before initiating temperature programming. Normally, a retention
gap (70 cm to 5 m of uncoated, deactivated fused silica capillary) is placed be-
tween the injector and the separation column. This focuses the less volatile com-
ponents and minimizes band broadening of alkane peaks eluting during tempera-
ture programming [24]. If this mode of injection is used without a retention gap
to separate waxes, then there is a risk of peak broadening and splitting.
This injection technique is very useful in the analysis of refined and synthetic
waxes in that it requires dilute sample solutions (minimizing the solubility
problems). Also the technique has proved to give a reasonably linear response
for alkanes up to C78 [25] and has been used to separate polywaxes up to C120
chain length. It can be used to quantitatively analyse waxes (with components of
wide boiling range), giving accurate and precise carbon number distributions.
However, there are several problems inherent in using- this technique:
1 . Due to limitations in minimum needle size available, the smallest column
that the sample can be injected into has to be at least 0.32 mm internal di-
ameter. If a retention gap is used, then the separating column can be of
smaller diameter. However, for high temperature work, the columns have
to be joined by a "Graphpack" type connector (which can be a source of
problems).
2. The thin silica syringe needles used are difficult to handle and keep clean.
It is possible that a small amount of sample will move back along the sy-
ringe by capillarity, or be transferred from the outer syringe wall and
smeared along the inlet wall of the column. To guard against this, it is
recommended that there should be a minute air and solvent gap between
the sample and the syringe plunger, as well as an air gap at the syringe tip.
This method of sample introduction is most accurate when a stainless
steel needle is used in the automated mode. Precision engineering and mi-
croprocessor control have now made automatic on-column injection a re-
liable technique.
3. Residues can build up in the injection zone leading to contamination of
later samples, particularly if the column is used at a higher temperature
than usual. This part of the system needs regularly removing, which is
easier if a retention gap is used.
The programmable temperature vaporizer (PTV) injector was developed by
Poy [26] and Schomburg [27] into a practical method that could be used in the
split, splitless (on-column) or solvent flush mode of injection. In a modern PTV
injector, a sample solution is injected, via a septum, into a narrow glass tube
containing glass- or quartz-wool, connected to the capillary column. The sample
condenses onto the wool insert, which is initially kept at ambient temperature.

62 Chapter 3
Then the injector body is rapidly heated so that each sample constituent is
eluted, in turn, onto the column inlet (which is 10-20C below the boiling point
of the solvent) where it condenses. After an initial period in which all the sample
components are transferred to the column, the oven temperature program is
started.
This technique avoids the interaction of the stationary phase with the injected
sample, associated with the splitless and on-column injectors. Also, after flash
vaporization, the solvent and sample are condensed as a narrow band within a
short distance of the column inlet. This optimizes the solvent effect and leads to
quantitative separation over a wide molecular weight distribution. There is no
need for a retention gap with its associated connector problems at high oven
temperatures. However, there can be a deposition of components in the split
outlet and septum purge if they are not heated or insulated correctly. These re-
quire regular cleaning with solvent followed by oven drying.
The advantages of the PTV injector are:
1. Any size of capillary column can be easily fitted into the glass insert.
These range from wide bore columns for rapid analysis of polywaxes, to
the latest microbore columns used to obtain detailed research information
regarding wax blends.
2. An ordinary domed tip syringe with a tapered needle (which does not
damage the prepierced high temperature septum during injection) can be
used. This minimizes the chance of septum pieces being flash vaporized
onto the column, causing ghost peaks. The problems associated with silica
needles (i.e. backflushing and needle contamination of the column) do not
arise and the system can be easily automated.
3 . The sample components vaporize sequentially, reducing the need for a
large volume glass injector liner (which is required for classical splitless
injection) and decreasing sample flashback, septum contamination, peak
splitting and solvent tailing to a minimum [28]. Also any involatile mate-
rial collects on the glass or quartz wool insert, which can easily be
changed.
4. It is a versatile injector, being capable of split, splitless and solvent purge
modes. Therefore, a wide range of samples can be catered for. In the split
mode the response factor for alkanes appears to be linear over the whole
boiling point range. As there is a definite difference between the boiling
points of the solvent and the sample components, the solvent purge mode
can be used for large volume injections of dilute solutions.
The disadvantages of the PTV injector are:
1. Dirt can accumulate in the insert and this needs to be regularly changed to
eliminate the occurrence of ghost peaks.
The chromatographic analysis ofrefined and synthetic waxes 63
2. If the PTV injector is used in the splitless mode, it is important that the
vaporizing chambers is not too small to take the large volume of solvent
vapour from a 1p1 injection. Otherwise the solvent vapour can flow
backwards into the septum region or the carrier gas supply. It has been
recommended that the vaporizing chamber should have an internal vol-
ume of 1 ml [29]. Another alternative is to initially vaporize most of the
solvent at a low temperature with the split valve open, then close the
valve for the sample vaporization stage [13]. However, this reduces the
solvent effect.
Comparison of on-column with PTV injection (both split and splitless) for the
quantitative analysis of alkane standards up to C44 shows little difference in the
relative response factors. Initial comparative investigations between the two in-
jection techniques used to separate compounds above C60 indicated that the
PTV in the splitless mode gave significant losses of higher boiling components
[ 181. This was later attributed to opening the PTV split valve too soon after in-
jection (i.e. before complete vaporization of the high boiling components) [30].
Hinshaw and Ettre [311 investigated this effect. They used Polywax 655 to show
that the PTV and on-column injectors can give equivalent recoveries of alkanes
up to C78 if the PTV split valve is initially closed for 4 min (although later re-
search found even this time too short).
Between 1985 and 1987, Barker carried out extensive research into quantita-
P * c k
t u' x J
Fig. 3.2. Capillary GLC analysis of typical rubber wax blend. Column: 25 m x 0.32 mm i.d.,
0.17 ,um cross-linked methyl silicone, 95 kPa He, PTV injector 370C. Temp. prog. (1) 60C for
5 min; (2) 7C m i d to 360C.

64 Chapter 3
tive HTGLC. This included investigating the best operating conditions for using
the PTV injector for separating refined and synthetic waxes [13,32]. A PTV in-
jector was used up to 450C in the split, splitless and solvent flush mode to sepa-
rate alkanes to above C70 (Fig. 3.2). It was found that the PTV had to be set to
20C above the maximum column temperature for 4 min to ensure that all the
components of any hydrocarbon wax were transferred onto the column (which
was kept at 60C for this stage). To improve this situation additional insulation
was placed around the injector body (particularly around the split outlet from the
injector body). For quantitative analysis in the splitless mode, the split valve was
opened only after 9 min. For the analysis of dilute samples of synthetic waxes,
the solvent flush mode was used. Initially the solvent is removed through the
split valve (using a PTV temperature of 80C for 1 min) before vaporizing the
components onto the column. This allowed the injection of a large sample vol-
ume without the solvent tail affecting the baseline in the region of the first com-
ponent peaks.
This work was amplified and extended by Tipler and Johnson [33], who used
Polywax 500 and a microcrystalline wax to investigate the optimization of a
PTV in the split mode. They found that, as well as PTV temperature, the PTV
heating period definitely affects mass discrimination, peak shape and peak dis-
persion (for Polywax 500 an optimum of 20 min was required). The importance
of only opening the split valve after a significant time (at least 10 min) and other
effects (detector temperature, make-up gas, temperature program and carrier gas
flow) were investigated. Recently, a practical guide to using the PTV has been
published [34]. Ai Instruments, Cambridge, have produced a PTV injector ca-
pable of use up to 600C. This now enables all the components of Polywax 1000
to be rapidly eluted onto a high temperature column without discrimination [35].
3.2.2.2 Detection
The flame ionization detector (FID.) is commonly used for the analysis of
waxes. However, the new atomic emission detectors has been used for the GC
analysis of polymer additives [36]. Mass-spectroscopy could also be used, but is
limited by the maximum temperature of the transfer line from the GC, the maxi-
tnum mass number detectable by the instrument and by the expense. The FID is
mass flow sensitive and the response of hydrocarbons is proportional to the
number of carbon atoms passing through the flame at a given time. For quantita-
tive analysis it is important that the FID conditions are controlled to give linear-
ity of response for all the eluted alkanes (i.e. no mass discrimination).
GC instrumentation designed for packed columns can be converted for capil-
lary GC. However, make-up gas (usually nitrogen) is required to obtain the best
flow for unbiased detection. For accurate quantitative analysis of high melting
waxes, the capillary GC FID requires to be designed for low flow at high tem-
The chromatographic analysis of refned and synthetic waxes 65
peratures. Adding make-up gas to the FID may improve the chromatography of
microcrystalline waxes [3 I].
The column should end near to the tip of the detector to minimize any high
temperature catalytic cracking on metal surfaces. It has been claimed [37] that, at
high FID temperatures, thermionic emission also takes place on the metal tip
leading to increased background noise. The same paper also suggests that the
higher boiling components of synthetic waxes may be adsorbed onto the metal
FID surface, leading to peak tailing. However, this seems to be improbable con-
sidering the high detector temperatures used. Some detectors now have ceramic
tips to overcome these problems.
The FID should be kept at least 10C higher than the maximum programmed
column temperature to avoid the higher boiling components from condensing out
on any detector surfaces. To eliminate condensation, it is also important that the
detector is well insulated so that there are no cold spots before the FID jet. Most
FID detectors are only designed to reach a maximum temperature of 450"C, even
although the associated GC oven can be programmed to 500C. The practical
temperature program maximum is 440C. In 1988, Carlo Erba produced the Wax
Analyser with an FID detector designed for HTGC, and a few other companies
have now followed their lead.
Wax materials are complex, containing many constituents that elute close to-
gether and are often not completely resolved. These rapidly eluting components
require a detector with a short time constant so that all the peaks are detected
without distortion. A time constant of 0.1 s or less is required to separate waxes.
Signal filtration is then important to remove detector noise [38], particularly
when it is used at high temperatures.
3.2.2.3High temperature GLC columns
The only limitation to the GLC separation of refined waxes and synthetic
waxes is thermal cracking, which starts under normal atmospheric conditions at
400C [39,40]. At 500"C, up to 90% of an alkane component in a wax may de-
compose (the thermal stability of alkanes decreases with increase in molecular
weight). However, as GLC analysis uses hydrogen, helium or nitrogen, this ef-
fect is reduced and the maximum practical separation temperature of alkanes is
probably between 450 and 500C.
The main problems associated with the HTGLC separation of alkanes are the
stability of the stationary phase and the column during temperature program-
ming. Above 250"C, packed column non-polar stationary phases start to degrade
and above 300C, bonded non-polar stationary phases on capillary columns tend
to degrade. This column bleed is reduced by deactivating the surface of the sup-
port material (which has to be a high purity silica) and by using a thermally sta-
ble stationary phase. High temperature non-polar bonded phases can now be

66 Chapter 3
used to greater than 450C. Metal capillary columns were used originally to
separate high boiling petroleum fractions, but high stationary phase bleeding
above 350C limited their use. These were followed by pyrex columns, but the
impurities in the glass led to a rapid increase in stationary phase bleed above
400C. Simultaneously, fused silica columns (made of high purity silica) were
introduced, externally coated with a polyimide. Polyimide coating has been im-
proved over the years so that fused silica columns can now be used up to 420C.
However, columns coated with aluminium have now extended working tempera-
tures beyond 440C.
During the early 1980s, the major advances in column technology were made
by those researching into simulated distillation techniques. In 1982, Szafranek et
a1 investigated the separation of high boiling crude oil alkane fractions using
Dexil 300 coated on a pyrex capillary column, with mass spectrometry detection
1411. They tested the thermal stability of this type of column to 410C and es-
tablished that the separation for each alkane below C30 was in the sequence
branched chain hydrocarbons before straight chains, followed by cycloalkanes.
In 1984, Luke and Ray [42] used a short pyrex column, coated with a thin film of
OV-1, to separate Polywax 500 up to n-C70 by programming the GC oven to
400C. It was found that above 400"C, the stationary phase bleed levels in-
creased rapidly, until at 420C decay occurred. This technique was developed
further by Munari et al. [18,43]. They coated a thin film of dimethyl silicone
onto 5-10-m long, wide-bore pyrex columns to separate Polywax 655 up to n-
C 120 at 430C.
Pyrex columns tend to lose stationary phase at an exponential rate as the col-
umn temperature reaches 400C. This is due to the trace metallic oxides in the
pyrex causing instability at the stationary phase interface. With high purity fused
silica columns this effect is removed and any bleed problems are mainly due to
instability in the stationary phase itself. Although fused silica columns and
chemically bonded stationary phases were introduced in 1979 [44,45], it was not
until 1986 that wax chromatography methods using these columns above 350C
were reported.
Perkin Elmer published [46] the results of an investigation into the use of
their PTV injector, combined with a 25 m x 0.25 mm i.d. fused silica column
(coated with a 0.1 y m methyl silicone bonded phase). The GC was temperature
programmed to 3 70C to qualitatively separate microcrystalline waxes into
straight and branched chain alkanes up to approximately C55, and total alkanes
to C68. However, the results were not much of an improvement on the work
carried out 13 years previously by Tulloch [9], who used a short column packed
with Dexil 300 and a temperature program to 400C to separate alkanes to the
same carbon number. Above 370"C, the standard polyimide coatings tend to
thermally degrade leaving small bare patches of fused silica, causing the capil-
The chromatographic analysis of reJined and synthetic waxes 67
lary column to disintegrate. Few standard stationary phases could be used in

fused silica columns above 370C at this time.
At this point, Lipsky and Duffy published their work on the manufacture and
use of aluminium-coated hsed silica columns containing a new bonded phase,
capable of application up to 450C for a limited time [47,48]. As the fused silica
column was drawn from its furnace, it entered a molten metal coating chamber.
On cooling this gave a smooth, bonded coating of aluminium approximately
20,um thick. A new bonded methyl polysiloxane phase and coating procedure
was developed to enable separations to be carried out up to 450C. They used a
15 m x 0.2 mm i.d. column coated with a 0.1 p m bonded phase to separate
Polywax 655 to above C90. A 1986 Quadrex Update Bulletin showed the same
type of column, but only 8 m long, being used to separate Polywax 1000 up to
approximately C 120.
There were initial problems with this technology, due to columns becoming
brittle after continuous use and spontaneously breaking. Improvements have
been made in the coating process so that the columns are less likely to disinte-
grate (only tending to break if repeatedly heated and cooled at a rapid rate). SGE
have developed a carborane-modified polysiloxane stationary phase which cross-
links on being coated inside an aluminium clad fused silica column [19]. Useful
work was carried out in investigating the changes in gas velocity up to 450C for
0.22, 0.32 and 0.53 mm i.d. columns. It was found that, at 70 kPa hydrogen pres-
sure, an optimum flow of 50 cm/s at 100C produced maximum resolution over
the whole temperature range. These stable HT5 series of columns are capable of
continuous operation at 460C. They have been programmed to 480C to sepa-
rate Fischer-Tropsch wax (near to theoretical maximum temperature for the
GLC analysis of alkanes). SGE have shown that the 0.53 mm i.d. columns can be
used to separate Polywaxes and the 0.32 mm i.d. columns can separate mi-
crocrystalline wax. However, as with most HTGLC column research, no atten-
tion has been given to the temperature program and flow conditions for quanti-
tative analysis.
Barker found that high quality polyimide columns could be externally condi-
tioned, by slowly increasing the maximum column temperature, so that the
polyimide further cross-linked. In this way, it was possible to regularly tempera-
ture program a 15 m X 0.25 mm diameter polyimide-coated column to 400C for
accurate quantitative analysis of microcrystalline waxes and some synthetic
waxes [13]. Chrompack U.K. then announced their new high temperature poly-
imide column, coated with a high temperature methyl silicone stationary phase
[49]. This was produced as an alternative to the metallized columns for HTGLC
up to 450C.
In comparison to aluminium coated columns, these columns were claimed to
be less fragile after repeated use at high temperature. AIso it is claimed that

68 Chapter 3
aluminium coated columns tend to work loose in ferules (due to the expansion
coefficient). Aluminium coated columns are conductive, therefore they have to
be carefully placed in the FID (or a silica column placed between the aluminized
column and the detector). It is also impossible to judge the internal quality of
used aluminium coated columns, whereas polyimide columns are transparent.
However, above 40OoC, polyimide hardens and becomes brittle, giving it a lim-
ited life-time. These factors are important in making a final, practical choice of
column.
A recent development in column technology has been the re-introduction of
metal capillaries, following on from the work of Takayama et al. [50]. They pro-
duced a metal column coated with a polydimethyl siloxane with hydroxyl end
groups, capable of use up to 430C (although it bleeds heavily when used above
350C). Buyten er a/. [51] improved the technology by coating a deactivated
stainless steel metal surface with a series of bonded layers, culminating in the
high temperature stationary phase (see Fig. 3.3). This research resulted in the
Chrompack Unimetal columns [52],which can be continuously used for 32 h at
450C with little effect on the capacity factors. They were designed for simu-
lated distillation use (e.g. the separation of Polywax 1000 up to 450C). How-
ever, a 25 m X 0.25 mm column coated with a 0.1 p m SIM. DIST. phase has
been used to separate slackwax to 375C. Metal columns used for HTGLC are
now a serious alternative to both aluminium and polyimide-coated columns.
In 1992, the Restek Corporation introduced a new type of HTGLC column
(Fig. 3.3). It was produced by depositing a thin film of fused silica onto the in-
side of stainless steel tubing followed by a thin deactivating polymeric layer,
which is then coated with a bonded high temperature stationary phase. This pro-
duces a tough, inert, thermally conductive column that can be continuously used
above 400C. A paper by Schuyler et al. 1531 showed that, as wide bore col-
umns, they can be used to separate Polywax 655 and 1000 up to 445C. A
0.28 mm i.d. column coated with a 0.25 ,urn non-polar high temperature phase is
available, but nothing has yet been published on the use of these columns for
quantitative wax analysis.
METAL TUB
PRIMARY CO
FUSED SILI
NTERMEDIATE
EACTIVATION LAYER
CHEMICALLY BONDED PHASE
Fig 3 3 . Cross-sections of current metal columns. (a) Chromopak unimetal column; (b) Restek
MXT column
Desty et al. [54] were the first to use long open tubular columns to carry out
high speed separation of petroleum products. Since the early 1980s there has
been an interest in microbore columns (ranging from 0.18 to 0.02 mm. i.d.) for
high speed GC [S].In 1988, MicroQuartz introduced polyimide coated fused
silica microbore columns, from 0.02 mm internal diameter upwards, for use with
high temperature stationary phases [56]. They are claimed to endure continuous
use for 80 h at 400C to 10 h at 450C. Although there are many advantages in
using these columns, there are also problems associated with their practical ap-
plication. The advantages are:
Increase in column efficiency with decrease in internal diameter. A 10 m
x 0.10 mm i.d. column may have up to 100 000 theoretical plates. Highly
complex microcrystalline waxes can be resolved to the baseline, leading
to accurate quantitative analysis.
Increase in sensitivity, because of the sharp peaks obtained. This means
that low solubility synthetic waxes can be injected at lower con-
centrations, reducing injection problems.
Decrease in column bleed due to the small amount of stationary phase in
the column. This reduces baseline problems at high temperature to a
minimum.
High speed analysis, which reduces the retention time of components.
With narrow bore columns, the theoretical plate height (H) varies little
with increase in linear velocity (u). Therefore, high carrier gas velocities
can be used to reduce the elution temperature of high boiling point al-
kanes (lowering the maximum program temperature of the column).
Compatible with GCMS systems due to the low carrier gas flow rate.
The disadvantages- are:
1. High component concentrations overload the column capacity. Therefore
more solvent has to be used to prepare the dilute solutions required for
injection.
2. The narrower the column diameter, the more difficult it is to fit into the
injector and detector. For on-column injectors there is a large drop in di-
ameter from the 0.32 mm diameter retention gap to the microbore column.
This leads to connector problems and extra-column band broadening. A
solution to this problem has been the invention by Hagman and Roeraarde
of the at-column injector [57], where the injection is made into a small
cavity directly above the column entrance.
3. The swift elution of many highly resolved peaks requires a detector and a
data handling system that can cope with the large number of rapidly
eluting components.
It is only recently that microbore columns have been used for HTGLC. Dam-
asceno et al. [58] evaluated their own borosilicate microbore column (6 m X

70 Chapter 3
0.1 1 mm) up to 390C, combined with on-column injection. They separated out
the porphyrin fraction of a Serpiano oil shale. Microbore columns have great
potential for separating microcrystalline and synthetic waxes. A PTV injector
with a narrow insert is probably the most effective injector. The flow through the
detector is obviously much reduced and requires make-up gas.
3.2.3 Quantitative gas liquid chromatography separation of waxes

Over the past 10 years there has been a steady growth in understanding the
quantitative GC separation of refined and synthetic waxes. Several papers have
been written on the use of the carbon number distribution of waxes [3-51. A pa-
per by Case [59] graphically illustrates carbon number distribution, the chroma-
tographic conditions that can be used to separate microcrystalline and synthetic
waxes, and their use in hot melt adhesive formulation. In these papers, no expla-
nation is given of the chromatographic separation or the means of obtaining ac-
curate carbon number distribution.
Nakagawa et al. [60] tried to characterise a whole range of waxes using solid
vaporization injection at 550C onto a capillary column coated with Dexil
300GC. Their pioneering work on carbon number distribution analysis unfortu-
nately suffered from the then common problems of injection cold spots and high
split flow ratios. These problems caused distortion of the high molecular weight
response factors for Polywaxes. However, their analysis of paraffin waxes and
correlation of these results to physical properties were of a high standard.
In advancing new injector or column technology to separate materials to the
highest carbon number, most authors have neglected chromatographic integrity.
There has been little attempt to obtain baseline separation of n-alkanes. Also it is
questionable whether some of the higher carbon number peaks reported for
Poljwax 655 actually exist [61]. It is feasible that the high boiling components,
on injection, partly condensed out in the cold on-column injector only to be
eluted when the oven was hot enough to heat the injector. Thomson and Rynaski
[62] observed that, during the chromatography of Polywax 655 using a IOm
wide-bore column, baseline perturbations occurred during the final isothermal
period giving many more apparent peaks than occur with a shorter column.
These additional peaks they attributed to column bleed produced by oven
cycling during the final isothermal period. The interpretation of HTGLC quanti-
tative analysis and the calculation of carbon number distribution are probably the
most complicated chromatographic procedures.
Much of the basic information leading to accurate quantitative wax analysis
has been gained from simulated distillation research (see previous sections). This
type of analysis does not require high resolution, whereas the quantitative anal-
ysis of refined waxes needs detailed separation of the branched and straight
The chromatographic analysis ofrefined and synthetic wares 71
chain alkanes. Short wide-bore thick-film columns can be used for quantitative
analysis of synthetic waxes (e.g. Polywaxes) if the simulated distillation tem-
perature program is adjusted to obtain baseline resolution. But the quantitative
analysis of refined wax materials requires capillary columns that are long and
narrow with thin films of stationary phase to sufficiently resolve the main com-
ponents.
The prime factors in optimising the separation of waxes are:
1. sample solvent;
2. mode of injection and detection;
3. type and treatment of the column support material;
4. stationary phase used;
5. stationary phase film thickness;
6. carrier gas purity, type and flow;
7. column size (length and internal diameter);
8. temperature conditions of oven, injector and detector;
The first four factors have been covered in previous sections, but there are
some practical points to obtaining precise and accurate HTGLC analysis. Par-
ticular attention should be paid to flushing the syringe with sample before injec-
tion, the technique of injection, and thoroughly rinsing it out afterwards. This
reduces the chances of contamination and leads to m accurate quantitative
analysis. The nearer that manual injection mimics aut J ~ J lted
- injection, the better
the precision and accuracy. With HTGLC, it is imif ant that the injection
points are regularly checked for cleanliness (i.e. on-column, retention gap or
PTV insert, and the injector components kept clean or changed). Otherwise
higher boiling components can build up in the system and be eluted with later
injections. If a septum is used, then it should be of the highest quality and regu-
larly checked for leaks. Any air leaks into the system during HTGLC will in-
crease the column bleed and distort the baseline, as well as reducing the col-
umns useful life. Therefore all connections should be regularly leak tested.
The carrier gas used affects most of the other listed factors in some way or
other. In HTGLC, it is not just important that a clean, dry and oxygen-free car-
rier gas is used, but it also has to be the most practical carrier gas for the separa-
tion required. Nitrogen has commonly been used to separate waxes by packed
column GLC [12], but van Deemter plots show that helium and hydrogen are
much better mobile phases for capillary GLC. Hydrogen has a flatter van Deem-
ter curve than helium and a higher optimum flow rate can be achieved. Compo-
nents separated with this gas can therefore be eluted with shorter retention times
and at lower temperatures during temperature programming. This is very impor-
tant for the separation of higher boiling components. However, the use of hydro-
gen is considered by some to be dangerous and helium may be the preferred gas
(particularly for quality control).

72 Chapter 3
The accurate control of the carrier gas by a back pressure regulator or mass
flow controller is also important for reproducible peak retention times. Flow
controllers have been strongly advocated for HTGLC analysis to theoretically
give an even resolution over the whole HTGLC temperature program range. No
account was taken of the substantial increase in gas viscosity (37% for helium
between 150 and 250"C), or changes in optimum velocity over the wide tempera-
ture range. Also mass flow controllers output constant volumes at ambient tem-
perature which expand at higher temperatures, considerably increasing the gas
velocity. The author has always thought that pressure control i s correct for wax
chromatography. At constant pressure, the gas velocity should slowly decrease
with increase in temperature so as to keep the resolution of the n-alkanes rea-
sonably constant (providing the optimum velocity and temperature program rate
was chosen) [32].
In 1990, Grob and Tschor [63] investigated the average carrier gas velocities
producing maximum separation efficiency at 370C. They used a 25 m X
0.32 mm i.d. column coated with 0.1 5 p m of a methyl-polysiloxane to compare
the van Deemter curves at 370C and 60C. Their results show that the optimum
gas velocities decrease with increase in temperature (approximately 35% be-
tween 60C and 370C). Also the difference in optimum gas velocities of hydro-
gen and helium is negligible for the columns normally used for wax separation
(around 20 cm/s at 370C). They proved that, at constant pressure, the decrease
i n gas velocities with increasing column temperature coincides with the decrease
in optimum velocities, whereas, at constant flow, the gas velocity increases away
from the optimum. Therefore constant pressure controllers are preferable for
wax analysis, and helium or hydrogen can be used.
The choice of column dimensions for quantitative analysis depends on the
wax being analysed and the degree of resolution required for accurate quantita-
tive analysis. Decreasing the column diameter from 0.53 to 0.10 mm greatly in-
creases the number of effective plates per metre (i.e. narrow columns have far
greater separating power). However, the capacity of the column decreases with
decrease in internal diameter, so that sample size must be reduced. To obtain
high resolution at high temperature a thin stationary phase film is used (0.1-
0.25 ,pni thickness); this further reduces the column capacity. The length of the
column affects the analysis time and the resolution. If the column is too long, the
higher molecular weight components suffer from peak broadening and eventu-
ally are undetected.
Short wide bore columns (8 m x 0.53 mm i.d.), made either of stainless steel
or aluminium-clad silica, can be used for Polywax separation. However, this size
of column is inadequate for resolving other synthetic waxes such as the Fischer-
Tropsch waxes. These waxes require the resolution of a 15 m X 0.32 mm i.d.
column. The new stainless steel columns are probably very appropriate for sepa-
rating these materials. Paraffin waxes and most wax blends can be efficiently
separated by a high quality 25 m X 0.32 mm i.d. polyimide-clad column. Wax
blends with large amounts of high melting point wax require a 0.25-0.20 mm i.d.
column, 15-25 m long to obtain adequate resolution at high temperature. High
melting point microcrystalline waxes require high temperature microbore col-
umns to resolve all the n-alkanes. The separation of a microcrystalline and syn-
thetic wax is shown in Fig. 3.4.
110 20 30 40 50 60 M i n s
b
C30
1 C40
I
30 40 50 60 Minutes
Fig. 3.4. (a) Capillary GLC separation of synthetic wax. (b) Capillary GLC separation of microcrys-
talline wax. Chromopak 15 m X 0.15 mm HT, temp. prog. 6W20C, 6C m i d .
References pp. 9&93

74 Chapter 3
For accurate quantitative analysis of both synthetic and refined waxes, a high
degree of resolution is required. This is not only obtained by the choice of col-
umn parameters and carrier gas flow, but also from the temperature program
conditions selected. With modern GC instruments, it is possible to accurately
control all the column oven conditions to obtain optimum resolution. Wax chro-
matography requires a slow temperature program to obtain the required resolu-
tion; this sometimes leads to analysis times of over 1 h.
The initial temperature of the GC oven is governed by the solvent used, and
should be set low enough to condense the sample solution in the column en-
trance. An initial short isothermal period is required for injection and the forma-
tion of a narrow sample band in the column. The oven can then be rapidly pro-
grammed (20-3O0C/min) to a temperature just below the first components elu-
tion temperature. This varies according to the sample, the column used and the
carrier gas flow rate. Another short isothermal period stabilizes the column tem-
perature before the sample separation stage of the temperature program.
For quantitative analysis, the separation stage has to be slow enough to
resolve the n-alkane peaks as near as practicable to the baseline. A tempera-
ture program rate between 4 and 8C is commonly used. Ideally the final
oven temperature is set so that the last peak has, if possible, eluted [13], but
if this is not feasible, then the final isothermal period should not be much
longer than approximately 5 min, otherwise the final peaks will broaden and
merge with the baseline. The elution temperature of individual hydrocarbons
depends on the ratio of the column radius to the film thickness (i.e. the phase
ratio), and the column length. This is the reason behind the choice of a short,
wide bore, thin film column for the high temperature separation of high mol-
ecular weight synthetic waxes. (Although narrow bore, thin film high tempera-
ture columns are required to resolve microcrystalline waxes for quantitative
analysis.)
Between 1985 and 1987, the author carried out research into the carbon
number distribution of wax blends [13,32]. This showed the importance of
knowing the response factor of alkanes of higher carbon number, as they vary
depending on injection technique and chromatographic separation conditions.
Several authors have used pure alkanes up to C44 to illustrate the differences in
response factor of different injection techniques. However, only a few authors
[ 25.62,64] carrying out simulated distillation research have clearly shown that
pure hydrocarbons can be used to measure alkane response factors up to C87.
Pure alkanes up to C60 [65] can now be purchased, which means that GC
equipment can be calibrated for response factors for the analysis of most refined
waxes. Polywaxes are used to empirically calibrate equipment for higher carbon
number responses, but there is a need for an internationally recognizable stan-
dard for HTGLC.
3.2.3.I Carbon number distribution analysis

Having successfully separated the wax, the next problem is to accurately
analyse the complex chromatogram. This type of quantitative measurement is
commonly known as carbon number distribution analysis. It sets out to quantify
the amount of each straight-chain (or normal) alkane, as well as the groups of
branched (or iso-alkanes) and cyclic alkanes of the same carbon number. For
each carbon number, this is simplified by measuring the peak area of each pre-
dominant normal alkane and separately totalling the preceding smaller peaks
back to the previous n-alkane to give the theoretical iso-alkanes of the same car-
bon number [40]. The results can then be plotted to give straight, branched and
total carbon number distribution graphs. The ratio of straight to branched chain
alkanes can also be obtained which is characteristic of the particular type of wax.
Wax chromatograms are complex and many GC methods do not resolve peaks
down to the baseline. Therefore, there are several ways in which the peak areas
can be measured. The theory and general practice of integration is covered else-
where [66], but a more detailed explanation is required to carry out accurate car-
bon number distribution analysis. Historically, the development of this type of
analysis has arisen from a need by rubber manufacturers for a detailed specifica-
tion for wax blend additives (using packed columns). The lack of both chroma-
tographic resolution and a reproducible GC method made interlaboratory agree-
ment difficult. This could only be accurately achieved by using a reference col-
umn and wax blend, so that the laboratories concerned could adjust the GC
conditions to obtain the same resolution. Also the method of integration had to
be clearly stipulated to obtain reproducible results. The three main methods of
packed column integration, which are the basis for capillary GC, are skim base-
line (following only the main troughs), valley-valley baseline (following every
trough) and horizontal baseline [ 131.
Barker investigated the accuracy of these different methods of integration us-
ing a standard rubber wax blend containing paraffin and microcrystalline waxes
[ 13,321. Using a modified form of the I.P. Method 372/85, the valley-valley and
skim baseline integration results were found to be similar, but the horizontal
baseline results differed markedly. It was noted that the baseline hump was con-
trolled by the degree of resolution obtained by the chromatographic method, and
that the maximum detectable carbon number increased with increase in tempera-
ture (reaching a maximum at 375C). These factors all affected the carbon num-
ber distribution and the ratio of total straight to branched chain alkanes. Inde-
pendent molecular sieve analysis showed that the experimental blend contained
approximately 25% branched chain alkanes. This was found to be similar to the
horizontal baseline result, but very different to the valley-valley and skim base-
line results. The work was repeated on a 15 m X 0.25 mm i.d., 0.1 p m DB-1 col-
umn (J&W) programmed to 370C. The wax blend (containing 5% heptadecane
References pp, 9&93

76 Chapter 3
internal standard) was injected into a PTV, which was programmed to 390C.
Electronic background subtraction was used together with the horizontal base-
line integration method, which gave total branched chain alkanes of 30%. From
this work, the horizontal baseline method appears to give the most accurate re-
su It s.
Michalczyk [67] published a paper on the quantitative analysis of paraffin
waxes, using cool on-column injection onto a 15 m X 25 m id., 0.1 p m OV-101
column. The GC oven maximum temperature was 330"C, whereas the detector
temperature was set at 300C. This leads to condensation of the higher alkanes
eluting from the column and distorts chromatographic results. A hexadecane in-
ternal standard was used and the background was subtracted electronically. He
gave detailed proposals for two methods of area measurement (see Fig. 3.5).
(a) For the first method, a horizontal line is drawn from just before the first
peak to the other side of the chromatogram, forming the baseline. The
main alkane peaks are identified, then perpendicular lines are dropped
from the troughs either side of the peaks to the baseline to enclose the n-
alkanes. The areas between the n-alkane peaks were designated as iso-
alkanes and cycloalkanes, taking into account that the iso-alkanes of a
given carbon number elute from the column before the n-alkanes.
(b) For the second method, the n-alkanes are integrated as if they were sitting
on top of an unresolved hump, above a horizontal baseline. The unre-
solved area of the hump underneath each of the n-alkanes is added to the
total iso-alkane area for the next higher carbon number.
Fig. 3.5. Carbon number distribution integration methods.

Method (b) was preferred by Michalczyk on the grounds that the total
branched chain alkane figures for this type of integration were (for the paraffin
waxes analysed) more comparable to those obtained by the urea adduct and the
molecular sieve methods for separating branched chain alkanes [68,69]. How-
ever, the second, higher melting paraffin wax gave anomalous results. There are
problems with obtaining accurate results with the urea adduct method. It has also
been shown that the molecular sieve method can be unreliable, due to variations
in the amount of branched chain alkanes separated by different batches of mo-
lecular sieve [70]. Therefore, these alternative separation methods are dubious
criteria for justifying an integration method.
The ASTM D.02 Committee has recently produced a draft method for the
capillary GC separation of petroleum waxes [71]. This method uses internal
standardization (by adding n-hexadecane) with cyclohexane as solvent. A variety
of column types and operating conditions are proposed for separating petroleum
waxes. Interlaboratory tests have been carried out to examine the repeatability
and reproducibility of the method. However, problems have been highlighted
and the method is under review.
The development of sophisticated integrators and PC software capable of
storing, reintegrating and redrawing chromatographic data, has enabled the car-
bon number distribution calculation to be more carefully examined. It is now
possible to rapidly collect peak data, subtract the column bleed baseline, exam-
ine the integration points in detail, and alter the integration conditions to obtain
accurate quantitative measurements. As wax chromatograms are complex with
many peaks, it is important to collect data at a rapid rate (10-20 data points per
peak) [72]. The column bleed baseline increases rapidly above 300C. It needs to
be stored as raw data [18] or as an algorithm [73] so that it can be subtracted
from the wax chromatogram to give the true sample baseline. Then the peak de-
tection parameters should be set to accommodate the different peak types (large
n- and small iso-alkanes, plus changes in peak widths over the whole chroma-
togram) [74]. Finally, the most contentious issue, the method of measuring the
peak areas (peak integration mode) has to be chosen.
With most paraffin waxes and for synthetic waxes, it is possible to set the
chromatographic conditions such that all the n-alkanes are resolved to a horizon-
tal baseline. Under these circumstances the horizontal baseline, baseline-
baseline, or the Michalczyk integration method will all give the same carbon
number distribution results. However, with intermediate and microcrystalline
waxes (or blends of them with paraffin waxes), the n-alkane peak resolution de-
creases and the different integration methods give divergent results. This is par-
ticularly true at the high molecular weight end of the distribution where it can be
difficult to distinguish the n-alkane peak areas from the iso-alkanes. In this
situation (see Fig. 3 . 9 , it is important that the position of the integration baseline

78 Chapter 3
and the integration points are clearly marked to establish the exact boundaries of
each peak area measurement. The question then arises as to what is the most ac-
curate way of measuring the area of a major peak when it is overlapped by minor
peaks.
This has been recently addressed in detail by Dyson [66]. For most wax
chromatograms, the n-alkanes are the major symmetrical peaks and the appended
iso-alkanes are much smaller, narrower asymmetric peaks. For these conditions,
the error of measuring the area of the smaller peak by dropping a valley perpen-
dicular increases as it becomes more of a shoulder on the side of the n-alkane
peak. However, if the integration software is written to construct a curving tan-
gential skim, then the error is much reduced. Where the overlapping peaks are
symmetrically shaped, and of the same height and width, then a perpendicular
line can be dropped from the valley to the baseline to enclose the areas. The
valley should be no more than 5% of the peak height. This condition can occur at
the end of the chromatogram. From recent work carried out by the author [75], it
appears that a combination of perpendicular and curved skim lines to a horizon-
tal baseline does accurately demarcate each n-alkane from the group of iso-
alkanes of the same carbon number (see Fig. 3 3 ~ ) ) This. is providing that the
chromatographic conditions are adjusted overall to obtain the best n-alkane peak
symmetry and resolution to baseline.
After identifying the peaks with alkane standards and measuring the peak ar-
eas, each group of iso-alkanes of the same carbon number needs to be totalled
and reported separately from the n-alkanes. If this is carried out manually using a
calculator, it takes a long time, but a PC with the requisite software can save
many hours of laborious work. It is possible to convert the binary data output
from a GC (or from an A/D converter connected to the GC) into ASCII text files
on a PC. These files can then be imported into a spreadsheet [76] (e.g. Lotus 1-2-
3 , QuattroPro, Microsoft Excel) and made into a table of n-alkane, iso-alkane
and total alkane results. From this table, a carbon number distribution graph can
easily be constructed (see Fig. 3.6). This procedure can be automated using a
spreadsheet macro or a specially written program, which makes the whole proc-
ess enjoyable! However, it is always essential with this complex type of analysis
to check the results against the chromatogram, particularly for mistakes in cali-
bration or calculation.
3.3 SUPERCRITICAL FLUID CHROMATOGRAPHY OF WAXES
The subject of supercritical fluid chromatography (SFC) in the petroleum in-

dustry is covered in another chapter and its use for high molecular weight petro-
leum products has been covered previously [77]. But for waxes, the importance
The chromatographic analysis of rejhed and synthetic waxes 79
4.5
3.5
0 2.5
5
$ 2
1.5
0.5
CARBON NUMBER
_ - -
Fig. 3.6. Carbon number distribution of a rubber wax.
and the difficulties of this type of chromatography deserve further comment. In

the mid-l980s, SFC showed great promise as a possible means of resolving high
molecular weight waxes into individual components, because it appeared not to
be restricted by sample volatility. It was thought that SFC would act as a bridge
between the high resolution capillary GC separation of paraffin waxes and the
inadequate resolution of synthetic waxes by size exclusion chromatography.
However, the development of HTGLC in the late 1980s has paralleled SFC ad-
vances and has undervalued the use of the latter for wax separation.
For wax analysis, SFC is a more complex method than HTGLC. To obtain an
even resolution over the whole carbon number distribution, a combination of
pressure or density, and temperature programming is often required. Using mi-
crobore columns it is possible to obtain a resolution somewhere between packed
column and capillary GC.
Sample injection is more complicated than for HTGLC, as a much higher
concentration of wax is required for the split injection system used. This initially
led to difficulties with solubility in the mobile phase and biased quantitative re-
sults. These problems have been largely overcome by the use of heated loop in-
jectors with rapidly rotating valves, which give a minute volume on-column in-
jection. Outlet restrictors have also been improved to give much more accurate
quantitative analysis. The column outlet is normally coupled to an FID, but there
is no reason why evaporative light scattering detection could not be used [78].

80 Chapter 3
Although SFC analysis does not achieve the resolution of HTGLC, its value lies
in being an alternative method giving another insight into the separation of high
molecular weight waxes.
The initial investigations into the separation of paraffin waxes were mainly
carried out using 5 0 p m diameter fused silica columns varying in length up to
20 m [79,80]. The preferred supercritical fluid was carbon dioxide (combined
with pressure or density programming), normally combined with FID. Split in-
jection was usually used at ambient temperature, which produced a tailing off of
the response factors for alkanes above C30. The later introduction of heated in-
jectors minimized this effect. Another common problem was the production of
electronic spikes, which is thought to be due to involatile solute particles enter-
ing the detector 1791. It was also found difficult to obtain an even, high resolu-
tion over the whole carbon number distribution. These last two effects can
clearly be seen in Fig. 3.7.
Some very advanced packed capillary column SFC research has been carried
out by Hirata and Nakata [81]. Using a small heated solid injector at 120C and
pressure programming, they successfully separated Polywax 500 to approxi-
mately C80 and Polywax 655 to C94. Their excellent work in separating syn-
thetic waxes was not improved until the late 1980s [82]. A typical separation of
Polywax 655 is shown in Fig. 3.8. Hawthorn and Miller managed to separate
a Fischer-Tropsch wax to above CIOO, but only resolved the n-alkane peaks
[83]. It has recently been shown that it is possible to separate Polywax 1000 up
I
I
.J.
0 10 20
Fig. 3 . 7 . Capillary SFC analysis of a rubber wax blend. Column, 10 m x 5 0 p m DB-5; oven temp.,
140C; mobile phase, carbon dioxide; pump, Brownlee microgradient system; injector, Valco
C14W l0Oul loop at 55C; FID detector, 350C.
The chromatographic analysis ofrefined and synthetic waxes 81
0 10 20 30 40
Fig. 3.8. Capillary SFC analysis of Polywax 655. Chromatographic conditions as before except:
elution conditions 2000-6000 psi in 25 min; total run time, 40 min.
to C130 using asymptotic density programming of carbon dioxide over 70 min

1841.
Advances in equipment and techniques have proved that it is possible to ob-
tain high resolution of alkanes to above C44 [85], with an even alkane response
factor to C60 [62]. However, it has not been possible to match the resolving
power of HTGLC to produce accurate carbon number distributions of both n-
and iso-alkanes for microcrystalline waxes. Several comparisons of simulated
distillation by HTGLC and SFC have been made (one for microcrystalline wax
[77] and one for Polywax 750 [62]). From these papers it can be concluded that,
by using a heated timed split injector, it is possible to obtain virtually the same
molecular weight distribution curve as by HTGLC.
3.4 SIZE EXCLUSION CHROMATOGRAPHY
Size exclusion chromatography (SEC), also known as gel permeation chroma-

tography (GPC), has been used for nearly 30 years to separate polymers accord-
ing to molecular size. The separating column (usually made of stainless steel) is
tightly packed with particles (up to 20pm diameter) containing pores of a con-
trolled size. A mobile phase is chosen that dissolves the sample, but is compati-

82 Chapter 3
ble with the column packing and the chromatography conditions. Sample mole-
cules that are larger than the pore size of the stationary phase pass through the
column at the same rate as the solvent. Smaller molecules diffuse into the pores
and are retained longer on the column; the smallest molecules being retained for
the longest time. Usually several columns are used to obtain the required resolu-
tion. They are packed with either mixed pore size material, or each column con-
tains a different pore size packing to cover the molecular range of the sample
components. The calibration and data handling have to be specially designed to
produce a molecular weight distribution curve that is the conventional way
round.
There are many books on SEC in general, but only a few which relate to low
molecular weight polymers such as waxes [86,87]. These books adequately
cover the theory, equipment and general practice of SEC. Therefore, only the
relevant points relating to wax chromatography are mentioned. SEC is a low
resolution separation method. For most hydrocarbon waxes, it produces a chro-
matogram consisting of a single curve with no discrete peaks (except for mixed
alkane standards where single peaks will be seen). The curve envelope contains
the sum of all the branched and straight chain alkanes merged together in a dis-
tribution from low to high molecular weight. The resolution of the curve depends
mainly on the size exclusion separation process, but it is also influenced by dis-
persion processes within the chromatography equipment (molecular diffusion,
column packing, etc.).
3.4.1 Early work on the SEC analysis of waxes

The standard procedures for analysing waxes using SEC were established be-
fore 1980. A few years after the establishment of the technique in 1964, it was
used to investigate cross-linked products from the irradiation of long-chain al-
kanes [88,89]. Salovey et al. carried out an excellent piece of research using a
series of polystyrene gel columns at 135"C, with trichlorobenzene mobile phase,
to separate alkanes up to C72. It is unfortunate that their alkane calibration up to
C94 (including response factors) cannot be repeated currently, due to lack of
synthesized standards.
In I97 1. Hillman produced an important comparative paper on the GPC and
GLC separation of waxes [90]. He used a variety of Styragel columns and sol-
vents, with refractive index detection. Tetrahydrofuran (THF) was used at 30C
to separate paraffin waxes, although these materials are more suitably analysed
by GLC. The RI detector response for alkanes, with this solvent, varies widely
and at C10 the response inverts [91]. THF has a maximum operational tempera-
ture of 4OoC, at which it requires careful handling (due to its toxic and unstable
nature), and is inappropriate for separating microcrystalline and synthetic waxes.
Toluene was used at 70C for separating all waxes, but was found to be insensi-
tive when combined with the RI detector for polyethylene waxes. However, at
that time, the preferred mobile phase was ortho-dichlorobenzene, combined with
an inhibitor, which was used at 80C.At high temperature, the chlorobenzenes
are highly toxic, therefore the equipment has to be placed in a fume cupboard
and the solvent has to be disposed of astoxic waste. The carbon number range
and maximum carbon number for both the GPC and GLC analysis of paraffin
and microcrystalline waxes were very similar. Several other papers on the GPC
separation of waxes were written around this time, but they were all based on the
previous papers.
Before the 1980s, porous silica was used, but became unpopular due to the
possibility of non-size exclusion interactions taking place. The Styragel columns
used by HiIIman and others were long and wide (120 cm X 7.8 mm). They
contained a large particle sized packing (37-75pm diameter) which gave typical
efficiencies of 3000 plates. This was equalled later by shorter columns (30 cm
long) containing smaller, 10p m u-Styragel. These columns were coupled to-
gether in a carefully selected combination (or column set) of 60, 100, 200,
400,500 and 1000 A pore size columns. The selection of each pore size depends
on the wax being analysed and the linearity required for the calibration curve
obtained by combining the columns. For the analysis of waxes, the columns and
injector should be placed in a thermostated oven. A long length of stainless steel
tubing has to be placed in the oven, between the pump and the injector, to pre-
heat the mobile phase.
The component parts of the SEC system were usually connected with the
minimum amount of stainless steel tubing (usually 0.25 mm bore). As mentioned
above, ortho-dichlorobenzene(with an antioxidant) was the preferred solvent for
wax analysis, because of its solvation power and compatibility with columns at
high temperature. The solvent was normally pumped at a flow rate of 1 ml min-I,
using either a syringe or a reciprocating pump to give a constant, pulse-free flow.
Before 1980 it was quite common to inject large volumes (up to 2 ml) onto the
column set via a septum, even although this increases the mobile phase viscosity
and sample band width. With the reduction in column particle size, injections
between 5 and 2000pl were made using four port, loop injectors. Automated
injection valves had been introduced by this time but were not commonly used.
Automation and a small sample size reduces band spreading.
The most common detector used in this work was the refractive index (RI)
detector. As electronics and other ideas were developed over the years, so the
detector was improved. However, it lacks sensitivity for waxes and its response
for a homologous series of alkanes can vary dramatically for some solvents
(even going negative) [91]. Also as the working temperature of the RI detector is
raised, its instability increases. In 1975, Dawkins and Hemmings used a modi-

84 Chapter 3
fied infrared detector, with a heated cell, for the high temperature analysis of
polyolefins [92]. Infrared detection is limited by lack of sensitivity and by the
shortage of suitable solvents with an absorbance window at the appropriate
wavelength to detect hydrocarbons. During this period, several experimental de-
tectors were used; these included the Pye Unicam moving belt and FID
transport detector, and the evaporative light scattering detector (ELSD), or
evaporative mass detector which was introduced in 1966 [93].
The detector signal was generally sent to a chart recorder and an integrator.
The integrator would be programmed to carry out a peak area slice routine and
calculate the mean molecular weight. Most SEC researchers in the oil industry
only had access to these basic tools, and calibration had to be carried out
manually. However, a few SEC research teams in other areas did have access to
mainframe computers to calculate the baseline drift, carry out area counts, store
calibrations and calculate the separate molecular weight parameters. It is fortu-
nate for wax chromatographers that calibration can be carried out using stan-
dards of similar molecular size. The pure monodispersed alkanes, and Polywaxes
of very narrow molecular mass distribution were well used as calibrants during
this period. The use of highly characterized broad molecular mass calibrants was
introduced by the US National Bureau of Standards (NBS) in the early 1970s.
However, it was not until 1978 that a low molecular weight, linear polyethylene
standard (NBS SRM 1482) was produced covering the upper range of synthetic
waxes [94].
3.4.2 Present day technology

The precise electronic control of pumps, ovens and detectors, plus advances in
detector design, have made SEC data more accurate and reproducible. Develop-
ments in column technology (i.e. the reduction in particle size and column di-
ameter, plus the development of mixed pore size packings) have enabled waxes
to be separated efficiently. Data handling, using powerful PCs and sophisticated
software, has allowed the analyst to measure and view the data in far greater de-
tail than previously. Also, after a moribund period between the mid-1970s and
mid-l980s, the status quo of high temperature SEC and low molecular weight
polymer analysis is now being questioned. There is a definite need for high
resolution SEC analysis of waxes, particularly for characterizing new synthetic
waxes and for investigating mixed polymer blends containing waxes (e.g. hot
melt adhesives).
By 1980, the use of a series of 25-30 cm long columns, 8 mm in internal di-
ameter, was common. Each column contained 1 0 p m diameter particles of dif-
ferent pore size. The columns were arranged to give the best calibration for a log
molecular weight versus retention time graph (typically a third order polynomial
calibration). In 1982, Waters introduced the Ultrastyragel columns with three

times the efficiency of the u-Styragel columns By the mid-l980s, 20 p m mixed
bed columns (each containing all the pore sizes used in a column series) were
commonly used in tandem to separate polyolefins. However, these mixed bed
columns did not have the resolution appropriate for microcrystalline and syn-
thetic wax analysis. Also the high temperature polyolefin analytical method
(using di- or tri-chlorobenzene eluent) is not appropriate for waxes or wax
blends, as the antioxidant peak in the eluent interferes with the low molecular
weight end of the chromatogram [95].
Following these improvements in packing materials, reliable 5 p m polysty-
rene-divinylbenzene phases, for individual pore size columns or mixed gel col-
umns, became available for low molecular weight analysis [96]. These columns
are much more efficient and have a higher resolution than the 10pm columns.
The Polymer Laboratories 5 p m Mixed-D column has an efficiency of >50 000
platedm and is linear over the molecular weight range 200-400 000. However, it
is important to reduce dead spaces to a minimum, e.g. by reducing connecting
tubing internal diameter and length, or the improvements will be lost in band
broadening. It is also important to use a guard column of the same material to
avoid column blockage at the injection end. Mixed bed columns can be com-
bined together to obtain higher resolution, without the calibration discontinuities
that occur when combining several different pore size columns.
In 1991 Polymer Laboratories introduced a 3 p m mixed pore size column [97]
for separating low molecular weight materials (with a linear calibration up to
30 000 molecular weight). These columns have an exceptional efficiency
(approaching 100 000 plates/m) and can be used for rapid analysis. However,
they can only be used up to 100C and require a system with low dead volume
and an accurate micropump to work effectively. Low dead volume is achieved
by using short, narrow connecting tubing and a low volume injector (i.e. lop1 or
less). A useful advantage is the lower solvent consumption, but the low flow rate
can lead to detection problems. No work has yet been published on the use of
these columns for wax analysis.
Several authors have tried to use packed SEC material in long capillary col-
umns [98]. MicroSEC, as it is called, is a very difficult technique and typically
can give an efficiency up to 50 000 platedm [99] (similar to the standard 5 p m
columns). These columns are also much more expensive than standard columns
[99]. The more practical options are the 4.6 mm i.d. columns (produced by Phe-
nomenex and Polymer Laboratories) and the 2.1 mm i.d. columns (produced by
Phenomenex). These columns are packed with 5 p m mixed pore size material.
Ying et al. [ 1001 used cyclohexane at 70C as a mobile phase to separate iso-
tactic polypropylene (after it had been dissolved in hot decahydronaphthalene to
destroy the crystallinity). This is a much less toxic material than chlorobenzenes
References pp. 9&93

86 Chapter 3
and has a lower viscosity. It has been claimed [87] that aliphatic hydrocarbons
shrink the column packing, causing voids and channels to form, and lowering the
efficiency. Ying et al. found to the contrary. Their u-Styragel columns were used
continuously with cyclohexane for more than 4 months, with no change in reso-
lution. Since then, the stability of styrene-divinylbenzene columns at high tem-
perature has been much improved.
Barker has used cyclohexane at 70C in a series of 10p m columns to success-
fully separate both refined and synthetic waxes [95] (see Fig. 3.9). This solvent
is difficult to use in conjunction with a refractive index detector, because the
refractive index difference between cyclohexane and waxes is very small and
therefore the sensitivity is low. Nor can it be used with an infrared detector as
there is no appropriate solvent window. However, it can easily be used, in con-
junction with the evaporative mass detector, to detect all waxes with higher
sensitivity.
The evaporative mass detector (or ELSD) is manufactured by several compa-
nies worldwide (Polymer Laboratories Ltd, UK; Varex Corporation, USA;
Sedex, France). Each is a different design, but they all work on the same princi-
ples [ l o l l . The column effluent is passed through a nebulizer and converted into
18 -
17 -
lfi -
15 -
14 -
13 -
12 -
11 -
10 -
09 -
08 -
L
0.0 I
0.04 0.05 0.06 0.07 0.08 0.09 0.10 0.11 0.12
Molecular Weight * 10e4
Fig. 3.9. Overlaid SEC analysis of refined and synthetic waxes. -, Polywax 500; ----, mi-
crocrystalline wax; - * - * -, Polywax 655.
a fine spray using pressurized nitrogen. The spray passes along a heated column
where the solvent is volatilized at a controlled temperature, leaving the non-
volatile solute to pass through a light beam (produced by a filament lamp or la-
ser). Light is scattered by reflection and refraction off the separated component
particles and is detected at 90 or 120 to the light source by a photomultiplier or
photodiode detector. If the experimental condition are optimized [102,103], the
output from the detector is proportional to the mass of eluent over the whole of
the molecular weight range. This makes it a sensitive, universal mass detector.
However, it is essential that the connection between the column oven and the
detector is as narrow and as short as possible. This tube should also be heavily
insulated and preferably heated, to minimize the plating out of the high mo-
lecular weight components on the inner walls.
Until recently, ELSD instruments did not have the same response factor for
alkanes over the whole of molecular weight range, with a tailing off of the re-
sponse for lower alkanes (due to their volatility). Also the signal of each compo-
nent may not be directly proportional to the concentration. Varex have produced
an electronic linearizer to overcome the latter effect for single component
peaks. Recently, Polymer Laboratories Ltd. have redesigned the evaporative
mass detector (Model PL-EMD 950) to address the problems inherent in all
previous instruments of non-linear response and concentration effects and have
greatly increased the sensitivity of the detector. This has been mainly achieved
by placing two fibre optic bundle collectors at an optimum angle of 120 either
side of the light source. They are connected to a remote photomultiplier tube.
Also a heated connection with the column oven has been produced. The detector
works best if the sample concentration is small (i.e. 0.001-0.20% of solute) and
the solvent evaporation temperature is as low as possible (vaporization depends
on latent heat not boiling point). These conditions are ideal for waxes, particular
the higher molecular weight synthetic materials which, at room temperature, tend
to be insoluble in many solvents.
A more sophisticated and expensive form of the ELSD is the multi-angle laser
light scattering detector (MALLSD). The column effluent is examined directly
in a specially designed flow cell. A laser light beam passes through the cell and
the scattered light is detected by an array of photodiode detectors arranged in a
circle either side of the light path. Using this arrangement, it is possible to meas-
ure absolute molecular weight and size distribution accurately. This detector was
originally restricted to higher molecular weight polymer analysis, but Wyatt
Technology Corporation, USA have now proved that it is possible to success-
fully use it for high temperature SEC and low molecular weight samples (i.e.
polystyrene molecular weight 760) [ 1041. However, it requires a concentration
detector in series as the sensitivity of MALLSD increases with higher molecular
weight. With waxes, this wouId normally be the refractive index detector, as a

88 Chapter 3
high sample concentration is required for the MALLSD. However, refractive

index is an inaccurate means of mass detection at low molecular weight due to
the changing response factor of the components.
Another detector which could be used is the viscometer, but this also requires
a concentration detector (usually refractive index). Viscosity detection has been
used since the early 1970s [105]. It was not until the mid-1980s that a sensitive
differential viscosity detector, that could be used with SEC, was produced by
Viscotek, USA [106]. In SEC, the molecules are separated by molecular size, i.e.
the hydrodynamic volume. This is equal to the sum of the intrinsic viscosity and
the molecular weight of the polymer. The calibration curve for the logarithm of
the hydrodynamic volume versus retention volume of all polymers is the same.
Therefore, with this detector it is possible to produce a universal calibration and
accurately measure the molecular weight of an unknown. This detector can also
be used in conjunction with known standards to plot a graph of branching fre-
quency versus molecular weight. Although the differential viscometer has been
used extensively for polyethylene analysis, it does not appear to have been used
for waxes, The mathematical relationship of intrinsic viscosity and hydrody-
namic volume with low molecular weight materials is more complex than at
higher molecular weights [ 1071. Also refractive index is an inaccurate means of
mass detection at low molecular weight. Therefore care is needed in using this
detector for analysing waxes.
Other detectors have been developed, but do not appear to be commercially
popular. In 1983, a flame ionization detector for liquid chromatography was pro-
duced by Tracor Instruments, USA [108]. This should have a linear mass re-
sponse for all alkanes. A thermospray interface coupled to a Fourier transform
infrared detector by a moving metal ribbon [lo91 shows promise, but has not
been commercialized.
The most promising recent innovation has been an infrared detection system
produced by Lab Connections Inc. [I lo]. This involves spraying the SEC eluent
onto the edge of a rotating germanium disc, where the solvent is removed, and
then analysing the deposited material by rotating the disc in an infrared spec-
trometer. Lab Connections have used this detector for high temperature SEC
analysis of low density polyethylene and for determining branching as a function
of molecular weight distribution. It shows great promise for wax blend analysis
where additives such as ethylene vinyl acetate are part of the composition. An-
other promising technique that is being increasingly used for LC separation, but
does not appear to have been used in SEC, is electrospray mass spectrometry.
Combined with high resolution microbore SEC, it should be possible to obtain
absolute data on molecular weight distribution and branching.
There have been great advances in data collection from detectors, PC hard-
ware capability and data handling software. This has opened up access to expert
systems for SEC data handling [ 1 111. Polymer Laboratories have been the leader
in using Microsoft Windows software to enable the analyst to write his own
sample data handling method, calibrate his instrument and view the results more
easily than before [ 1121. Fluctuations in flow rate between calibration and sam-
ple runs can be offset using internal standardization techniques, and the chroma-
togram is corrected accordingly. The analyst can now carry out post-analysis
examination, including overlaying chromatograms and exporting the data to a
spreadsheet [ 1121.
Calibration procedures have been clearly dealt with in publications already
cited [87]. In 1968, it was possible to carry out an SEC calibration using alkanes
up to molecular weight 1500 [88]. Today, only alkanes up to molecular weight
962 are available. Polymer Laboratories produce a series of Polywaxes and
polyethylene standards up to 14 000 molecular weight which can be used for
narrow range calibration of synthetic waxes. However, some are not ideal be-
cause they have a polydispersity (M, to M,) greater than 1.1. The author is at
present investigating the use of new synthetic waxes to extend the molecular
weight range for narrow standards. In the United States, NBS Standard SRM
1482 has been well characterized as a low molecular weight broad standard [94].
Broad standards are not readily available, as they require fractionation and char-
acterization of the components to produce a cumulative weight fraction table.
They are a single run standardization and are valid for non-linear calibrations.
With the advent of high quality mixed bed columns and detectors, plus the use
of accurate routine calibration, internal standardization and modem data han-
dling, it is now possible to obtain accurate and reproducible SEC chroma-
tograms. Apart from the standard log. molecular weight distributions, it is now
possible to view the linear molecular weight distributions (dwldMversus M) and
the cumulative linear molecular weight distributions [95]. This is very relevant
to the SEC of waxes where the molecular weight distribution is relatively narrow
(see Fig. 3.9). It now appears possible to carry out a quantitative SEC mass dis-
tribution analysis for waxes, similar to the GC carbon number distribution
analysis (but not with the same precision).
3.5 CONCLUSIONS
Instrumental and computer advances over the past 10 years have given the
analyst the tools to investigate waxes and wax blends in great detail. It is now
possible to use GC to clearly separate waxes into straight and branched chain
alkanes for quantitative analysis. However, there is still a need for a thorough
investigation into the correct method of peak integration. Hopefully the stan-
dardization bodies will resolve this issue in the near future. Harmonization be-

90 Chapter 3
tween wax blend producers and customers would then be easier and product de-
velopment more rapid.
SFC separation has not been popular, but SFE could prove useful for trace
additive and contamination analysis in waxes (particularly with increasing hy-
giene legislation). Recent advances in SEC column technology, detector design
and computer software have made it possible to obtain an accurate molecular
weight distribution of waxes, particularly when they are blended with other
polymers. However, there is a need for more standards between n-C60 and the
highIy characterized NBS polyethylene standards. Accurate SEC analysis of raw
materials and their blends, combined with other methods of analysis (e.g. differ-
ential scanning calorimetry and rheology), could be developed into a powerful
research tool. This would enable the materials scientist to understand raw mate-
rials and predict properties of blends more accurately.
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92 Chapter 3
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CHAPTER 4
Hydrodynamic chromatography of
polymers
Jaap Bos and Robert Tijssen

Amsterdam (Shell Research B. V.), Badhuisweg 3, I031 CM Amsterdam,
Koninklijke/Shell-Luboratorium,
The Netherlands
4.1 INTRODUCTION
Hydrodynamic chromatography (HDC) is nowadays a well-known technique,

which is mainly used for the separation of colloidal particles of different sizes.
Basically, particle size separation takes place when the finite dimensions of the
particles are large enough to interfere with the flow profile in a packed bed or
open tubular flow system. For geometric reasons, particles are excluded from the
slower moving wall regions in the flow channel and will therefore travel at a
velocity greater than the average velocity of the solvent. Hence, larger particles
elute first from the flow system, followed by the smaller ones, as is also the case
in the well-known polymer separation technique of gel permeation chromatogra-
phy (GPC), also called size exclusion chromatography (SEC).
The technique was pioneered by Small [1-31, who also coined the name HDC.
In his work, he demonstrated the practicability of HDC for the separation of
mixtures of polymer latexes and colloidal suspensions, using packed beds of im-
permeable particles as the separation column. Pedersen [4], Mori et al. [5],
Prudhomme et al. [6] and Hoagland et al. [7] showed that packed-column HDC
was a suitable technique not only for particle separations but in principle also for
the separation of large molecules such as proteins, polystyrenes, and water-
soluble polymers of high molecular mass (such as xanthanes and polyacryla-
mides). More recently, Kraak et al. [8] and Stegeman et al. [9] greatly improved
the separating power of HDC by using columns packed with extremely small
(1.2pm) non-porous silica particles.

96 Chapter 4
Mullins and Orr [lo], Noel et al. [ll], Brough et al. [12] and Shuster et al.
[ 131 demonstrated experimentally that HDC separations of differently sized par-
ticles can also be effected in a single open capillary tube with a diameter of the
order of several tenths of a millimetre. Elie and Renaud [ 141 succeeded in sepa-
rating fibrous particles in paper fibre suspensions using even wider open tubes of
15 mm internal diameter.
Although microcapillaries (defined here as having internal diameters less than
10pm) had already been used by Silebi and DosRamos [ 151 and DosRamos and
Silebi [16] for the separation of particles, Tijssen et al. [17,18] and Bos et al.
[ 191 specifically used these capillaries for the separation of polystyrenes and for
studying the dissociation behaviour of micelles of two-block polymers.
Only for microcapillary HDC has the theory been sufficiently developed to
calculate the size of an eluting species solely fiom its residence time relative to
that of a small molecule, without recourse to calibration standards. In capillaries
of a larger diameter, and in most packed-bed columns, the situation is more dif-
ficult, since exclusion fiom the wall is then not the only separating mechanism,
because other, only partly understood phenomena, such as tubular pinch and
molecular elongation, also strongly influence the transport mechanism through
the column. Moreover, in the separation of particles that are charged or can ob-
tain a surface charge, such as polystyrene latexes in water, there is a pronounced
influence of the electric double layer at the column wall on the migration
mechanism. In this case, ionic strength and velocity of the mobile phase are of
importance. Since in the context of Chromatography in the Petroleum Indus-
try, HDC separations of polymeric species are the more important ones, they
will be treated in detail. For a review of particle separations by HDC, the reader
is referred to articles by Dodds [20] and McHugh [21].
4.2 MICROCAPILLARY HYDRODYNAMtC CHROMATOGRAPHY
4.2.1 Theory
Throughout this chapter, a microcapillary is (rather arbitrarily) defined as a

column of circular cross-section with an internal diameter of less than 1Opm.
This size is sufficiently small to ensure that in normal HDC operation with the
flow of dilute solutions of macromolecules, extra effects such as tubular pinch
are absent. It can safely be assumed that the flow profile is laminar, i.e. a para-
bolic Poiseuille profile exists (see Fig. 4.1), where the local velocity u(r) for a
radial position r in a tube of radius R (0 2 r IR) is given by
U(r)=2u[l-(r/R)*] (4.1)
Hydrodynamic chromatography of polymers 97
I /
Fig. 4.1. Schematic representation of the laminar flow transport of a spherical solute particle
(diameter 2a) in Poiseuille flow through a cylindrical capillary (diameter 2R).
here the average velocity ii is

R
U = (2 / R 2 ) lu(r)r dr (4.2)
for point molecules that rapidly exchange between all available streamlines.
These molecules (e.g. the solvent molecules) emerge from a column of length L
after time
L
t m =-- (4.3)
24
Now, as is schematically depicted in Fig. 4.1, the centre of gravity of a parti-

cle or macromolecule of size 2a in the flow field cannot sample the low-velocity
regions near the wall. The average particle velocity lip is now
again provided that the diffusion process with coefficient Dm of the particles is
sufficiently fast to sample all the available streamlines in the time t of the trans-
port process, i.e. the dimensionless Fourier number Fo = DJR2 % 1. Combining

98 Chapter 4
Eqs. (4.1) and (4.4), and introducing the aspect ratio A = a/R, we find
zp=i7(1+213.-2) (4.5)
The average residence time for the particle is thus found from tp= L/iip, and the
dimensionless residence time z (compared to a small molecule) is now
t = t , I t , =(1+2A-A2)-1 (4.6)
Equation (4.6) is still rather idealized, because the polymer molecules have
been regarded as rigid, non-rotating particles. Actually, since the part of a mac-
romolecule that is closest to the capillary wall is in a region of a lower velocity
line than the opposite side of the molecule, the resultant couple of forces will
make the particle rotate, which influences the translational velocity. This hydro-
dynamic problem has been investigated theoretically by DiMarzio and Guttman
[22-241 and Brenner and Gaydos [25]. Tijssen et al. [18] showed that their re-
sults could be described by a modified quadratic term in Eq. (4.6):
t = (1 + 22 -a?)-' (4.7)
where C has a value of 2.698 for permeable macromolecular coils, while for im-
permeable hard spheres its value is 4.89.
The importance of Eq. (4.7) is that it shows that microcapillary HDC can be
used to calculate the size of an eluting species without recourse to calibration
standards. From the measured relative residence time, z, the aspect ratio, A, can
be calculated, and from this and the known diameter of the capillary, the size of
the particle can be obtained. Even if the conformation of the macromolecule is
unknown, and so the exact value of constant C in Eq. (4.7) is in doubt, the size
can be estimated with only a small error. By assuming that for random coil mac-
romolecules, C will have a value between the two extremes mentioned above
(C = 2.698 for macromolecules in a good solvent, and C = 4.89 for the limiting
case of an impermeable hard sphere), Bos et al. [ 191 showed in a practical ex-
ample for a separation with z = 0.95 that the sizes calculated with these two val-
ues differed by less than 4%.
A plot of 13. versus t,the calibration curve of microcapillary HDC, is shown in
Fig. 4.2 (Eq. (4.7) with C = 2.698 and 4.89). The curve is very steep close to
z = 1, where the exact value of C is not too important, but where a small error in
the determination o f t will lead to a large error in A, and hence a large error in
the calculated size of an eluting species. This is not the case at lower t values,
where the slope of the calibration curve is less, but in this region exact knowl-
edge of the conformation of the macromolecule or particle is needed to use the
Hydrodynamic chromatography ofpolymers 99
Aspect R a t i o , A
I00
Dimensionless residence time
Fig. 4.2. Calibration curves for microcapillary HDC (Eq. (4.7), 't = (1+U- C,E2)-') with C = 2.698
(random coils) and C = 4.89 (hard spheres).
correct value of C, and so to obtain meaningful values for the calculated sizes.
We do not think that the upper part of the calibration curves where the quadratic
term in Eq. (4.7) is the leading term, and where ever larger particles will elute at
ever longer residence times, has any physical meaning. Here, the aspect ratio il is
so large (>0.2) that the concept of a particle eluting in an undisturbed Poiseuille
profile that is used in the derivation of Eq. (4.7) is probably invalid. Mavrovoun-
iotis and Brenner [26] proposed a complicated theory to describe this case of
closely fitting particles.
4.2.2 Expressing the size of macromolecules
For macromolecules, there exist a number of different measures for their

mean size, e.g. the radius of gyration (rg),the hydrodynamic radius (rh),the end-
to-end distance, etc. In hydrodynamic chromatography, the size that we deter-
mine is yet another one, the effective radius (T), related but not equal to the ra-
dius of gyration. To appreciate the difference, let us look more closely at the
time scale on which these two measures are determined. The radius of gyration
is usually determined by dynamic light scattering, by the interaction of a photon
with a macromolecule. This is a very fast process and during this interaction, the
conformation of a given macromolecular coil is not changed. Averaging over the
conformation of a number of molecules will give the radius of gyration.

100 Chapter 4
By contrast, the interaction of a macromolecule with a wall is a slow process

compared with the time scale on which a given coil will change its conformation.
So now we average over a number of macromolecules that all have assumed
every possible spatial configuration. The resulting concept of the mean maxi-
mal cross-section, 2F, as introduced by Volkenstein [27] and further discussed
by van Kreveld and van den Hoed [28], determines the distance, r,by which a
random-coil macromolecule is excluded from the wall. Now T is related to the
radius of gyration by the following equation:
F = (& / 2)rg = 0.886rg (4.8)
The factor of 0.886 in this equation is, however, only valid for random coil
macromolecules in good solvents where internal rotation of the segments is pos-
sible, since the macromolecular coil is expanded by the interaction with the sol-
vent molecules. However, in a theta solvent, where, by definition, the expansion
of the coil by solventlsegment interaction is exactly compensated by intra-
molecular segmentlsegment attraction, the polymer chain is rather contracted
and behaves more as a rigid sphere with hindered internal rotation. In this case,
the coefficient in Eq. (4.8) will more probably approach a value of 1 [18]. For a
more detailed discussion on the size of polymer molecules, the reader is referred
to [29].
4.2.3 Instrumentation
4.2.3.1General aspects
The demands for the instrumentation that is needed to work with microcapil-
laries follow from the characteristic values of the chromatographic parameters
that are presented in Table 4.1 for a typical microcapillary HDC column that has
been used in [ 181:
TABLE 4.1
TYPICAL VALUES FOR A MICROCAPILLARY HDC COLUMN
Length (cm) 330

Internal diameter (um) 2.68
Votume (nl) 16
Head pressure (bar) 200
Volumetric flow rate (nl/min) 0.7
Length of detected zones (cm) -4
Volume of detected zones (pl) -230
From the values, we see that a standard liquid chromatograph is not suitable
for this type of analysis. High-pressure pumps that can deliver volumetric flows
of the order of several nl/min do not exist and a standard detector (with a flow
cell of say 1 ,u1) cannot accommodate peak volumes in the picoliter range. Also,
operation of such a column would require an injection size in the picoliter range,
another specification not met by standard liquid chromatographic injectors. All
this leads to a home-made instrumental set-up schematically represented in Fig.
4.3. A standard HPLC pump is used to deliver the mobile phase through a T-
piece at a volumetric flow rate of 2-3 ml/min. The major part of the solvent
flows through a restriction capillary dimensioned such that the required head
pressure on the microcapillary HDC column is attained. The length L and the
radius R (in cm) of the restriction capillary can be calculated from the Poiseuille
equation
where F is the volumetric flow rate (cm3/s), q is the dynamic viscosity (Poise),
and AP the required head pressure (bar). The sample is introduced into the mi-
crocapillary by means of an injector, the loop of which is only partly emptied by
actuating the valve for say 1-4 s. This ensures that the injected sample reaches
I D,,...* resistance capillary
Mobile phase vessel-
- Injector
Fig. 4.3. Schematic representationof the apparatus for microcapillaryHDC.

-
UV Defector

102 Chapter 4
the microcapillary as a rectangular pulse. Injecting the whole (or a partially

filled) loop will give badly tailing peaks in the highly efficient HDC columns
where, in theory, plate numbers of several hundred thousands are obtainable.
4.2.3.2 Detection
A most important part of a microcapillary separation technique, the detection
of the separated zones, is also the operation that strongly limits the practicability
of the technique. Sensitive techniques such as thermal lens detection are difficult
to apply, while others techniques, e.g. potentiometry and laser-induced fluores-
cence are not particularly suitable for the majority of the industrial polymers.
Laser-based refractive index detectors are not sensitive enough for use in micro-
capillaries, so the only technique left that currently has practical application is
on-column ultraviolet (UV) absorption. This technique is performed by remov-
ing part of the polyimide coating of the capillary and using the resulting trans-
parent section as a flow cell. Not all commercial UV detectors are equipped to
accommodate a microcapillary column, since the detector compartment must be
easily accessible to place the column into the light path and install movable knife
edges to block (most of) the stray light (i.e. light not passing through the capil-
lary in the very centre of the column). Even with these modifications, the poly-
mers under study should contain strong chromophoric groups. As an example,
with detection at 215 nm, a polymer should contain more than 10% weight of
e.g. aromatic rings to make detection with a sufficiently high signalhoke ratio
possible. Beneficial effects can be expected from the recent improvements in the
instrumentation for capillary zone electrophoresis (CZE). Although at present
the W detectors for CZE are designed for columns with an internal diameter of
50,um, they can, even without modification, be used for microcapillary HDC,
albeit with a signahoise ratio that is roughly five times poorer than that of the
detector used by the authors in [ 181. In view of the current rapid improvements
in CZE technology, it can be envisaged that in the near future, microcapillary
HDC will be performed with standard commercial instrumentation.
A sensitive detector is very important for microcapillary HDC separations,
since the injected concentrations of polymers cannot be too high to avoid con-
centration effects (entanglement between the macromolecular coils). For most
polymers with a relatively low molecular mass of say <2 X lo5Dalton (Da) a
concentration of about 1 g/f is sufficiently low; for higher molecular masses, it
may be as low as 0.1 g/l. Even then, microcapillary HDC can, to date, only be
used to study polymers of which the polydispersity D (D= weight average mo-
lecular masshumber average molecular mass) is not too large, say D < 1.3.
Polymers with a larger D will give elution profiles which on account of the ex-
cellent molecular mass separating abilities of microcapillary HDC, are very
broad and low, and difficult to detect with a reasonable signalhoise ratio.
Another detection technique that has been used with microcapillaries is the
use of a make-up flow at the end of the column. In this case, the volumetric flow
rates can be so high that a standard detector for microbore HPLC columns could
be used, but in view of the very large dilution of the eluting zones, a sensitive
technique like fluorescence detection is needed, which is not a viable option for
most commercial polymers. The make-up flow technique has, however, been
successfully applied by DosRamos and Silebi [161 in the separation of latex par-
ticles in a 4 pm internal diameter column, because in that case, the elution zones
were very narrow (and so the signals relatively high) due to the tubular pinch
effect, which we will treat later in more detail.
4.2.3.3 The column
For calculating the size of eluting species, the internal diameter of the micro-
capillary should be accurately known. Basically, there are two simple methods to
determine the average value of this diameter:
(1) The flow method at different pressure drops AP, accurately measured
with a pressure transducer, the average linear velocity ii is determined
from the residence time of a pulse injection of small molecules (e.g. tolu-
ene). From Eq. (4.9), it can be derived that a plot of ii versus AP should
yield a straight line with a slope of 106R2/8yL,from which the average
value of R can be calculated.
(2) The volumetric method the microcapillary is filled with toluene. After
this, the front end (with the restriction capillary disconnected) and the exit
side of the column are rinsed with acetonitrile. With the restriction capil-
lary in place, the toluene is flushed from the column into a known volume
(1-5 ml) of acetonitrile by pumping say at least three column volumes of
acetonitrile through the microcapillary. In this way, solutions containing
some mg/l of toluene are obtained, the concentration of which can be ac-
curately determined by, e.g. reversed-phase liquid chromatography with
ultraviolet detection at 205 nm. From this concentration, the internal vol-
ume V of the microcapillary can be calculated, from which the average
value of the column radius is obtained as R = d( V/nL).
From the description above we see that, apart from the detector, the instru-
mentation needed for microcapillary HDC is rather simple and inexpensive.
A practical point in HDC instrumentation relates to the coiled form in which
the capillary is generally used during the experiments. It is known that coiling
can give rise to secondary flow effects that disturb the Poiseuille profile. Accord-
ing to Tijssen [30] this disturbance starts to become important for residence time
distributions if the dimensionless group
Re*Sd, > 50 (4.9)

104 Chapter 4
where Re = 2UNv is the Reynolds number, Sc = v/D, is the Schmidt number

and ,Icis the coil ratio (internal diameter capillaqdcoil diameter), with v being
the kinematic viscosity. In microcapillary HDC, the condition of Re2Sdc< 50
can easily be met, as is shown in the following example, where the effective ra-
dius and the molecular diffusion coefficient for polystyrene in tetrahydrofuran
(THF) has been calculated using the experiments of Schulz and Baumann [31]
and Mandema and Zeldenrust [32] as
F = 1.23 X 10-5Mo,558 Olm) (4.10)
D,=3.45 x 104M-0.564 (cmVs) (4.1 1)
A typical microcapillary HDC experiment uses a column of 300 cm length with

an internal diameter of 3 pm. With an analysis time of 20 min, the average ve-
locity U = 0.25 cm/s. A typical diffusion coefficient D, would be 6.7 x
10-8 cm2/s (e.g. a polystyrene of molecular mass 3.8 X lo6 Da in THF eluting at
z = 0.90). With a value of 0.005 cm2/s for the kinematic viscosity and a coil di-
ameter of 10 cm, we calculate that:
R e = 1.5 x Sc=7.5 x 104, 1,=3 x
The group Re2Sd, has a value of 5 X lo4, so Eq. (4.9) tells us that no coiling
effects are to be expected. With larger diameter capillaries where the Reynolds
number and 1, are generally much higher, the effect can, however, be very pro-
found. We will come back to this aspect later.
4.2.4 Applications
An example of a microcapillary HDC separation is shown in Fig. 4.4. Separat-

ing these four polystyrene standards in THF in 25 min is, of course, not unique
since the standard GPC technique can easily perform this. But the advantages of
HDC over GPC are that it can work with solvents that are not compatible with
the GPC packing materials, and that it provides direct information on the size of
the eluting species. An example of this is given in Fig. 4.5. Here, again, poly-
styrenes are separated in THF, but also in diethyl malonate (DEM), which is a
theta solvent for polystyrene at 3 1-36C. As we see from Fig. 4.5, a polystyrene
of a given molecular mass elutes later in DEM than in THF, indicating, as might
be expected, that the size of the molecule is smaller in the theta solvent. The
size, calculated from the relative residence time z in the 1.342pm radius col-
umn, is given in Table 4.2, together with the size determined by light scattering
0.900 0.925 0.950 0.975 1.000

Dimensionless residence time , r
Fig. 4.4. Separation of four polystyrene standards and toluene by microcapillary HDC. Column
diameter 3.0pm, mobile phase THF. From left to right: polystyrene standards of molecular mass
1280,775,315 and 51 kDa, and toluene.
techniques (Table I1 in [18], U calculated from rg using Eq. (4.8)). To avoid un-
certainty in the exact value of the C-term in Eq. (4.7), we have calculated the
results only for the three lowest molecular mass polystyrenes, that elute within a
z region where the value of C is not important.
The table clearly proves the strength of microcapillary HDC; even with very
simple instrumentation, it is possible to obtain information on the size of mac-
romolecular species, without recourse to calibration standards.
TABLE 4.2
COMPARISON OF RADII (nm) OBTAINED BY HDC AND LIGHT SCA'ITERING
Molecular Radii by HDC Radii by light scattering

mass
Pa) THF DEM THF Theta solvent
127000 12.2 9.7 12.3 9.8

233000 17.8 13.5 17.6 13.2
41 1000 22.3 18.0 24.6 17.5
~

106 Chapter 4
Mol. Moss (Polystyrene)

SWIO~ r
* X
X X
Mobile Phose'
3c T H F * x
X Dlethyl rnolonote
I * x
*x
1051 I I 1 1 1 1 1 8 I I I
0 87 0 89 0 91 0 93 0 95 0 97 10 99
Dimensionless r e s l d e n c e t l m e . r
Fig. 4.5.Comparison of experiments on polystyrenes in THF and in diethyl malonate. Column

diameter 2.68 p m .
Other applications are found in the study of two-block polymers consisting of

hydrogenated poly(styrene-isoprene). In non-polar solvents at room temperature,
these polymers form micellar aggregates with the more polar polystyrene part of
the molecule in the core of the micelle. An example is given in Fig. 4.6, where
such a polymer (weight average molecular mass 92 000 Da, styrene content
35.8% d m ) is analysed in a 3.70,um internal diameter microcapillary in two
different solvents. In the polar solvent, THF, the polymer forms a molecular so-
lution; from the relative residence time t = 0.9855 an effective radius
-
I' = 13.8 nm can be calculated. In the non-polar solvent, n-decane, the polymer
elutes much earlier at t = 0.9496, leading to a radius of V = 52.8 nm, indicating
that the micellar aggregate has been formed. The present analysis can also be
extended to find the temperature range in which the micellar solution in n-
decane dissociates into a normal molecular solution. Although this process can
be studied by a number of other analytical techniques, e.g. light scattering, HDC
has again the benefits of simple instrumentation and ease of interpretation. The
results are shown in Fig. 4.7, where a number of HDC separations at different
temperatures are presented. It is clear that up to 80C, the polymer exists in the
micellar form, while at a temperature of 105"C, a true molecular solution is
formed, in which the size of the polymer is the same as in THF at room tempera-
ture. A potential complication in the interpretation of these experiments arises if
Toluene
I I I I 1 I
0.900 0.925 0.950 0.975 1.000 1.025
Dimensionless r e s i d e n c e time
Fig. 4.6. Hydrodynamic chromatography of a styrene-isoprene two-block polymer in two different

mobile phases at room temperature: (A) in n-decane, elution of the micellar aggregate; (B) in THF,
elution as the molecular species. Reprinted from [19].
shear degradation of the micelle in the column were to occur. This, however, is
almost certainly not the case due to the limited shear stress at the wall (<SO0 s-')
during the flow process.
For experiments like these, involving more or less polar species in a non-polar
solvent, it is necessary to deactivate the polar silanol groups on the fused silica
column wall by silylation. Details of this procedure are given in [191.
Another application of microcapillary HDC is the investigation of the mobil-
ity of the monomers of two-block polymers in their micelle. These micelles dif-
fer in character from the more well-known micelles of detergents in aqueous
systems. In water, the detergent forms a molecular solution up to a certain con-
centration, the critical micelle concentration. If more detergent is added, the
additional molecules build into micelles, but there is a rapid dynamic inter-
change between the detergent molecules in the micelles and those in the sur-
rounding solution. The case of the micelles of the two-block polymers mentioned
above is quite different. Here, it is not possible to dissolve the polymer at room
temperature in a non-polar solvent, but only at elevated temperatures, say 140C.
Only upon cooling down this molecular solution, are the micelles formed even-

I08 Chapter 4
1 I I I I I
0.900 0.925 0.950 0.975 1.000 1.025
Dimensionless r e s i d e n c e time r
Fig. 4.7. Temperature range of the dissociation of the micelle of a styrene-isoprene two-block
polymer in n-decane. Reprinted from [ 191.
tually. For a number of reasons, it has thus far been assumed that these micelles
are static, rigid structures at room temperature, without exchange between the
monomers in the micelle and those in the (sparingly concentrated) solution. We
studied this problem with microcapillary HDC in a 3.85pm internal diameter
column with micellar solutions of two different two-block polymers of molecular
masses 105 and 1.5 X los Da. In Fig. 4.8, it is shown that a room-temperature
mixture of the two micelles (effective radii 56.8 and 71.0pm) gives the two
peaks expected. If the solution is heated to 120C and cooled down to room tem-
perature, a mixed micelle with a radius of 63.2pm is formed. Any exchange, at
room temperature, of the monomers in the micelles and those in the surrounding
solution should thus show up as a gradual appearance of the mixed micelle. Fig-
ure 4.9 shows that this is actually the case; at 20"C, the two discrete micelle dis-
tributions slowly change into the single distribution of the mixed micelle. Al-
though the process is slow at this temperature, taking about 1000 h to comple-
tion, it takes only 24 h at 30C while at 40"C, the mixed micelle is completely
formed after just 1 h. Thus, these microcapillary HDC experiments show that the
micelles of two-block polymers in non-polar solvents are not static structures,
but that there is in fact a fairly rapid exchange between the monomers in the mi-
celle and those in the surrounding solution, even at about 40C below their dis-
sociation temperature.
1 I I I 1 I
0.900 0.925 0.950 0.975 1.000 1.025
Dimensionless residence time r
Fig. 4.8. HDC separation of micelles from styrene-isoprene two-block polymers in n-decane. (A)
Molecular mass of polymer 1.0 X lo5 Da. (B) Molecular mass of polymer 1.5 X lo5 Da. (C) Room
temperature mixture of (A) and (B). (D) Mixture of (A) and (B), after being heated to 120C for
2 h.
The last application we present shows that, apart from molecular species, mi-
crocapillary HDC is also suited to the characterization of small solid particles,
e.g. metal detergents. These metal detergents consist of a solid core of inorganic
material, covered with an organic ligand. Figure 4.10 shows the separation of
metal-detergent particles in n-decane in a 3.63 p m internal diameter column, the
material eluting at t = 1 being free ligand material not attached to particles. The
relative residence time t = 0.9940 for the peak top position leads to a calculated
radius of 5.5 nm for this material. However, we must bear in mind that for rela-
tive residence times close to unity, the ;Z versus t calibration curve according to
Eq. (4.7) is very steep, so a small error in the determination of t leads to a large
difference in A, and so in a large error in the calculated size. However, t deter-
minations in microcapillary HDC can be precise; in our case, a four-fold deter-
mination of t gave a standard deviation of 1.7 X lo4 t units, which leads to a
95% confidence interval for the radius of 5.3-5.8 nm. To attain this small stan-
dard deviation in t,we had to increase the analysis time to 90 min, instead of the
more common 20 min (note that at t = 0.9940 and at analysis time of 20 min, the

110 Chapter 4
979 h
478 h
I16 h
48 h
24 h
8 h
Oh
0.900 0.925 0.950 0.975 1.000 1.025

Dimensionless residence t i m e T
Fig. 4.9. Formation of a mixed micelle from styrene-isoprene polymers of two different molecular
masses in n-decane at 20C.
0.900 0.925 0.950 0.975 1.000 1.025

Dimensionless r e s i d e n c e t i m e r
Fig. 4.10. HDC separation of metal detergent particles in n-decane.

Hydrodynamic chromatographyofpolymers 111
difference in residence time between the particles and the toluene t = 1 peak
amounts to only 7 s). If we were to try to improve the (time) separation between
the two peaks by prolonging the analysis times to several hours, difficulties
might be expected; firstly, it is very difficult to ensure a constant pump perform-
ance over a long time period, and secondly, since the dispersion of the small
molecules at t = 1 in microcapillaries is mainly governed by axial broadening,
their peaks become progressively broader, lower and less easy to detect at very
low linear velocities.
Up to now the applications we have described are in the field of separations of
polymers in organic solvents. In principle, it is equally well possible to perform
microcapillary HDC in aqueous mobile phases. To this end, the silanol groups
on the silica surface have first to be derivatized with e.g. glycidyloxyalkylsilanes
to produce a Glycophase type column. Even then, separations of e.g. polysty-
rene sulfonates have to be carried out in phosphate buffers to diminish electro-
static interactions with the column wall. But these separations could not be de-
scribed by Eq. (4.7), so most probably there is still residual interaction with the
silica column material. This is not a problem for the separations obtained, but it
obscures the quantitative interpretation of sizes. Other, more successhl separa-
tions were performed by the authors on biopolymers such as hemoglobin and
ferritin. Since the polymers mentioned can easily and more efficiently be sepa-
rated by capillary zone electrophoresis, we do not think that, in general, aqueous
microcapillary HDC is a technique worth developing for the separation of bio-
polymers.
4.3 CAPILLARY HYDRODYNAMIC CHROMATOGRAPHY
4.3.1 From hydrodynamic chromatography to tubular pinch
Since Eq. (4.7) is dimensionless, we may expect that there are no difficulties
in going from microcapillary HDC to capillary HDC, i.e. there is no upper limit
to the internal diameter of the column that can be used. So, if we have particles
that elute at t = 0.90 in a 3 p m column, and we want to have particles 10 times
as large elute at the same relative residence time, we could simply take a capil-
lary that is 30pm in internal diameter. This is not true, however, as Eq. (4.7) is
derived assuming that the particles during the time of the chromatographic proc-
ess can sample all the different velocity lines that are available to them in the
parabolic Poiseuille profile. The particles can move through the different veloc-
ity lines by molecular diffusion, but as large particles have small diffusion coef-
ficients, and they are separated in wider columns, there is an appreciable chance
that this sampling of the different velocity lines will be insufficient.
Refirencespp. 125-126
I12 Chapter 4
In the theory section, the dimensionless Fourier number Fo = D,t/R2 was in-
troduced. Fo can be interpreted as the number of times a particle moves through
the column cross-section during the time t of the chromatographic process. If Fo
is much smaller than 1, it means that a particle that is introduced at a given radial
position in the column virtually stays on the same velocity line all the time.
Thus, particles introduced at the exact centre of the column elute at a velocity
twice as high as the average velocity, while particles introduced near the column
wall, where the velocity is close to zero, elute only after a very long time. Under
these conditions one, of course, cannot expect to make any useful separation. In
our practical applications we have found that Fo 1 10 is sufficient to ensure a
sufficiently high rate of radial mass transfer.
Now, when can we expect that this insufficient diffusion and so too low a
Fourier number will occur? As an example, we have calculated Fo for a number
of column and particle size combinations. The data, tabulated in Table 4.3, per-
tain to a column length of 300 cm, an analysis time of 20 min and polystyrene in
THF, eluting at t = 0.90.
We see from Table 4.3 that for a column somewhere between 10 and 30 p m in
diameter, the Fourier number will become smaller than 10. In the l00pm col-
umn, Fo is even lower than 1, albeit for the analysis of a polystyrene of ex-
tremely high molecular mass. But even with polystyrenes of lower molecular
mass, the Fourier number will be low, since the numerical value of the Fourier
number is largely determined by the R2 term in the denominator of Fo. If we take
the case of a polystyrene with a molecular mass of 3.8 X lo6 Da (Fo = 3500 in a
3 p m column), this polymer would only have a Fo = 3 in a l00pm diameter col-
umn (now of course with a larger value for the relative residence time; t = 0.996
instead of 0.90).
In the literature on capillary HDC, one often encounters columns as wide as
500 pm, in which particles of e.g. 25 p m diameter are separated in a few min-
utes. However, these large particles which have a very low diffusion coefficient,
are eluted under conditions in which Fo Q 1 . Separation is still established be-
tween particles of different diameter, but it will be clear that Eq. (4.7) is not
TABLE 4.3
Fo FOR DIFFERENT COLUMNPARTICLE SIZE COMBmATIONS
Column diameter Molecular mass Diffusion coefficient Fo

Pm) ( D a x 10) ((cm2/s) x lo9)
3 3.8 67 3500
10 29 21 102
30 190 7.4 3.9
100 1500 2.3 0.1
Hydrodynamic chromatography of polymers I13
applicable to these separations. Therefore, another mechanism must be respon-

sible for the separation in these wider-bore HDC capillaries. This only partly
understood phenomenon, called tubular pinch, is thought to consist of two op-
posite forces that work on particles with not too small an aspect ratio, which are
transported under conditions of a high reduced velocity ReSc. One of these
forces moves the particles away from the wall region; the other force transports
the particles away from the column centre. The net effect is that the particles are
concentrated into an annulus at a certain radial distance from the column centre,
the position of which depends, among other factors, upon the aspect ratio and the
reduced velocity. For a more extensive treatment, the reader is referred to Ti-
jssen and Bos [29].
An example of a tubular pinch separation is given in Fig. 4.1 1, which shows
the separation of a polystyrene of molecular mass 6.77 X lo6 Da in a 83 1-cm
long capillary of 38.5pm internal diameter, with THF as the mobile phase.
While the relative residence time according to Eq. (4.7) should be high, viz.
0.987, and close to the toluene peak (z= 1 indicator), we find a much lower t
value of 0.835 experimentally. The Fourier number in this experiment is only
0.73, so, on average, the molecules did not have the time to diffuse even once
through the column cross-section during the analysis. Still, instead of the dis-
torted, tailing elution profile that one would expect from experiments with such
-Toluene
polystyrene elution time

according t o HDC theory
(eq. 4.7)
I I I I I I
0 20 40 60 80 100
Residence time ,s
Fig. 4.11. Tubular pinch effect on a polystyrene sample [molecular mass 6.77 x lo5 Da) in a
38.5 pm wide capillary. Mobile phase: THF at 12 c d s .

114 Chapter 4
a low Fourier number, the tubular pinch effect gives rise to a reasonably sym-
metrical peak form.
We introduce 8 as the reduced relative residence time:
t(experimental)
6=
t(according to Eq. (4.7))
as the relative measure for the tubular pinch effect. If 8 = 1, then the experiment
is in agreement with Eq. (4.7), and no tubular pinch effect is present. In the ex-
periment mentioned above, we find a reduced relative residence time
8 = 0.835/0.987 = 0.846. Experimentally we find that, for fixed values of the
linear velocity and the column diameter, 8 is inversely proportional to the mo-
lecular mass of the eluted species; at fixed values of the molecular mass and the
column diameter 8 is inversely proportional to the linear velocity of the mobile
phase. Dimensional analysis reveals that 8 should be a function of the dimen-
sionless numbers Re, Sc and I. such that
8 =f(ReScA2) (4.13)
In Fig. 4.12, a plot of 8 versus ReSd* is presented for 219 experiments, in-
volving six polystyrenes (molecular masses 2.89 X 106-8.42 X lo6 Da) in five
columns of different internal diameter (7.53-38.5 pm), with THF as the mobile
phase. Column lengths varied between 200 and 835 cm, with mobile-phase linear
velocities ranging between 0.3 and 14 cm/s. The columns were used in a
straightened out horizontal position to avoid curvature effects (see Section
4.3.2). We see that all the measurements lie close to a single curve, indicating
that Eq. (4.13) is valid, although the exact form of the function f is still not
known (see, however, [29]). From Fig. 4.12, we conclude that for Eq. (4.7) to be
valid. a capillary HDC experiment must be conducted under conditions such that
ReSd2 5 1 (4.14)
At ReSd2 > 6, the functionfhas a high negative slope so, especially in that re-
gion, estimation of the particle size from a measured relative residence time t
alone, will lead to large errors.
It is stressed that tubular pinch effects are not necessarily found only in wide-
bore capillaries with high linear velocities. Even in microcapillary HDC, the ef-
fect can be found. For example, let us go back to Fig. 4.5, which shows the ex-
periments in a 323-cm long column with radius 1.342,um. The datapoint farthest
to the left stems from a polystyrene of molecular mass 3.61 X lo6 Da, that is
eluted in THF at a linear velocity of 0.28 c d s . Using Eqs. (4.10) and (4.1 l), we
can calculate a ReScjZ2 value of 4.7, indicating that the tubular pinch effect is
Hya'roa!ynamic chromatography ofpolymers 115
Reduced relative residence time, 6
0.90 $5 R
%r
09
Columns (10) Used:

0.85 - 0 PS 2M89 7.53 p m
0 0 d
0 PS 3M84
0 PS 4M48 11.7 p m v-
o PS 5M48
A PS 6M77 18.1 pm ?v
0.80 - V PS 8M42 2 5 . 0 pm T
VQ
38.5 prn
0 .75 I I 1 I 1 IlII I I I I 1 1 1 1 1 1 I 1 I I l l l j
lo-' loo 10' lo2

ReSCX2
Fig. 4.12. Tubular pinch effect in experiments on polystyrenes (six different molecular masses, five
different column diameters, in total 2 19 experiments).
already present in this typical microcapillary experiment. For ReSd2= 4.7, we

read an approximate 8 value of 0.99 from Fig. 4.12. This 1% deviation may seem
a minor effect, but it means that the experimental relative residence time of
0.8836, corrected for tubular pinch, results in a z value of 0.8925. When this
latter value is used in Eq. (4.7), it gives a radius of 89 nm for this polymer, close
to the expected value of 88 nm that can be calculated from Eq. (4.10). Using the
uncorrected value of 0.8836 in Eq. (4.7) would lead to an overestimation of
about lo%, viz. 98 nm, for this radius.
Now, how do we recognize whether and if in a capillary HDC experiment the
tubular pinch effect is present? Normally we do not know the diffusion coeffi-
cient or the radius of the polymers under study, so a calculation such as the
above cannot be made. In performing HDC experiments at different mobile
phase velocities, however, the tubular pinch effect is easily recognized. Since the
magnitude of the tubular pinch effect depends upon the linear velocity of the
mobile phase, an increase in the z value upon lowering the mobile phase velocity
indicates that tubular pinch is present. If the z value is invariant with the mobile

I I6 Chapter 4
phase velocity, then the experiment has been conducted under conditions such
that no tubular pinch occurs and Eq. (4.7) is valid.
If the HDC experiment i s an old one that we cannot or do not want to repeat,
or if we examine experiments from the literature, we still can make an estimate if
tubular pinch effects have not been present. To that end, we calculate the appar-
ent molecular radius r using the experimental t value in Eq. (4.7). If tubular
pinch effects are present, t values are shifted towards lower values, and the cal-
culated radius will be over-estimated, so r > F. From r, we calculate the appar-
ent molecular diffusion coefficient D, from the StokesEinstein equation
(4.15)
where k is the Boltzman number (1.38 X 10-l6 g cm2/s2K) and T is the absolute
temperature (K). Since ris over-estimated, D, will be smaller than the true, un-
known D,. As the Stokes/Einstein equation is in fact only valid for spherical
solid particles, application of Eq. (4.15) to a macromolecular coil will, in that
case, give an even lower apparent diffusion coefficient (compare e.g. Eqs. (4.10)
and (4.1 1) with Eq. (4.15) for polystyrene in THF to see this effect). Also,
A = r/R will be higher than the true aspect ratio A = r / R . Taking all this into
account, the apparent value (ReSd2) will be larger than the true value for
ReSd. Only if (ReSd2)< 1 can we be sure that there was no tubular pinch ef-
fect in the experiment.
As an example, let us calculate this for the polystyrene of molecular mass
3.61 X lo6 Da that was mentioned earlier as an example of tubular pinch in a
microcapillary HDC experiment (datapoint farthest to the left in Fig. 4.5). From
the uncorrected relative residence time z = 0.8836, we found the apparent radius
of 98 nm. From this, the apparent diffusion coefficient of D, = 4.51 X
10-8 cm2/s is calculated according to Eq. (4.1 5) (with 0.00488 P for the viscosity
at 22C). With 0.28 cm/s for the linear velocity and an internal radius of
1.342pm for the column, we calculate that (ReSd2) = 8.9. Since this value is
larger than 1, the experimental value for t is suspect; tubular pinch may well
have been present, although we do not know if the real R e S d 2 value is also lar-
ger than 1 (earlier, for this specific case, we calculated that R e S d 2 = 4.7, is in-
deed larger than 1,using the exact F and D, values from Eqs. (4.10) and (4.11)).
If we take the lowest left data point in Fig. 4.5, with a relative residence time
t = 0.9823, we can calculate that (ReSd2)= 0.017. This value is so much
smaller than 1 that there will have been no tubular pinch effect in the experi-
ment. Thus, it is permissible to use Eq. (4.7) to calculate the radius from the
relative residence time. This is, of course, confirmed by the result for this poly-
styrene of molecular mass 1.27 X lo5 Da, that has already been presented in
Table 4.2.
As a last remark on this subject, we want to point out that, although tubular
pinch has been identified as a separating mechanism that can operate at low
Fourier numbers, this does not mean that the tubular pinch effect is therefore
absent at high Fourier numbers. In our experiments leading to the construction of
Fig. 4.12, sometimes Fourier numbers in excess of 200 could be calculated. This
means that there is a rapid radial mixing of the polymer through the available
column cross-section, but still the relative residence time was lower than it
should have been according to Eq. (4.7). Consequently, tubular pinch is a rela-
tively strong effect, that can accumulate molecules into an annulus at a certain
radial position in a column, even when opposed by counteracting diffusion or
dispersion mechanisms that would tend to re-distribute these molecules through-
out the whole available cross-section.
4.3.2 Coiling effects
With the columns used to establish the 8 versus ReSd2 graph in Fig. 4.12, it
is possible to separate polystyrenes with different molecular masses, using the
tubular pinch effect as the separating mechanism in a single HDC experiment.
PS 2M89
I I I
I
75 90 105 I20 I35 I50

Residence t i m e , s
Fig. 4.13. Tubular pinch separation of two different polystyrenes in an 18.1 pm wide capillary.
Mobile phase: THF at 3.6 c d s .
References pp. 125-1 26

118 Chapter 4
An example of this is given in Fig. 4.13 for a mixture of two polystyrenes with
molecular masses of 8.42 X 1 O6 and 2.89 X 1 O6 Da in a column of 18.1 p m in-
ternal diameter, using THF as the mobile phase at a linear velocity of 3.6 c d s .
From the relative residence times (0.80 and 0.97), determined in a run in which
toluene had been added as the t = 1 indicator, it is easy to calculate that the sepa-
ration mechanism in this experiment is due to tubular pinch, since (ReSd2) is
much larger than 1 for these peaks ( 5 X lo4 and 52, respectively). Also, since in
this case we know the exact size of these polymers (Eq. (4.10)), it can be pre-
dicted that a HDC separation without tubular pinch would produce peaks that
would hardly be separated at all (relative residence times 0.97 and 0.98). Al-
though the separation is still incomplete, it should be noted that two polymers of
rather high molecular mass have been separated in less than 2.5 min.
It is clear that the chromatographic spreading of the polymers must be dimin-
ished to make such a separation more useful. To attain this, coiling of the HDC
column could potentially be of importance. As mentioned earlier, coiling distorts
the Poiseuille profile, an effect that becomes notable when the dimensionless
quantity Re2Sd, is larger than 50. Coiling has the effect of increasing the radial
Toluene
Cotled column
. . . . . . . . . Straight column
I I 1
40 56 72 88 I04 120
Residence time , s
Fig. 4.14.Effect of column coiling on the width of the elution profile of a polystyrene of molecular
mass 3.84 X lo6 Da in a 38.5pm wide capillary. Mobile phase: THF at 9.7 c d s . Coil diameter
6.3cm.
mixing in the column, which shows up as a narrowing of the chromatographic

zones [30]. An example of this is shown in Fig. 4.14 for a polystyrene of molecu-
lar mass 3.84 X lo6 Daytransported at a linear velocity ii = 9.7 cm/s through a
83 1-cm long capillary with an internal diameter of 38.5 pm. In a straight column,
the polymer elutes at z = 0.93, showing that there is a tubular pinch effect
((ReSd2) = 8000, ReSd2= 13). The peak has a standard deviation of (T =
10.6 s. When the experiment is repeated, but now with the column coiled (coil
diameter 6.3 cm), the polymer elutes at the same relative residence time, but it is
distinctly narrower with (T = 7.5 s. In this last experiment, Re2Sd, = 2300.
In a number of experiments in this coiled column with polystyrenes of other
molecular mass, we invariably found that the relative residence times were the
same as in the straight column. Thus tubular pinch concentrates the polymer in
an annulus in the column, and the strong secondary flow introduced by coiling is
unable to redistribute the molecules through the available column cross-section.
However, the increased radial transport of the molecules within the annulus
leads to a reduced chromatographic band spreading.
To diminish the band spreading to such an extent that the polymers in Fig.
4.13 are baseline separated would require much higher Re2ScA-, numbers. This
implies higher linear velocities in a longer column that is more tightly wound.
With the fused silica column used in the experiment of Fig. 4.13, this is impos-
sible, since the required pump pressure would be too high. Also, fused silica
capillaries cannot be wound in a very tight coil. With the wider metal and Teflon
columns that have been reported in the literature, this poses no problem, so in
those cases the best separations can be expected with tightly coiled columns that
are operated at the highest possible mobile-phase linear velocity.
4.3.3 Applications
Almost all the applications that are found in the field of capillary HDC are
separations of particles, e.g. latex, pollen, garnet, paper fibres, bacterial spores,
silica, paints, etc. The diameters of the capillaries used span a wide range of in-
ternal diameters (14-500pm). In the separation of paper fibres even a column
with an internal diameter of 15 mm has been used, which shows that the term
capillary is a rather flexible description. This wide range in column diameters
leads to various instrumental designs, where the small internal diameter experi-
ments are carried out in an instrument like the one described in Section 4.2.3 on
microcapillary HDC, while with larger internal diameter columns, standard
HPLC injection and detection techniques can be used. Normally ultraviolet ab-
sorption is used as the detection technique, although Zarrin and Dovichi [33] use
a sheath flow cuvette with a light-scattering detector.

120 Chapter 4
The applications that describe separations of polymers are rare. Revillon and
Boucher [34] separated a cross-linked polystyrene sample in THF in a 120-m
long capillary of 250pm internal diameter. Brough et al. [12] applied a 50-m
long, 242pm internal diameter column to separate a used engine oil into two
peaks, one of which was due to particulate debris such as carbon, metals and
polymeric products. They also analysed a THF solution of a water-based emul-
sion paint on a 50-m long, 450pm internal diameter column to fingerprint
successive batches of paint. Especially in the case of the analysis of the used
engine oil, we feel that the separation between the two peaks could have been
improved if a much smaller coil diameter than the reported 30 cm had been used.
Tazaki and Homma [ 3 5 ] used a stainless steel column (90 m long, 250pm inter-
nal diameter) and a fused silica column (25 m long, 100pm internal diameter)
for the separation of fluorescence-tagged xanthane polysaccharides in water and
aqueous buffers. As there is no possibility of calculating molecular dimensions
from the relative residence time alone, when tubular pinch is present, Tazaki ex-
presses the size as the equivalent particle diameter, i.e. the diameter of a poly-
styrene latex particle that would have eluted at the same relative residence time.
He states that his result, 330 nm, is in good agreement with the HDC experi-
ments in packed columns of Prudhomme and Hoagland [36] who found a value
of 150-300 nm.
In summary, examples of polymer separations in capillary HDC are not abun-
dant. Still, we feel that the method has potential for the characterization of high-
molecular-mass materials that are difficult to handle in GPC or packed-column
HDC. For efficient separations in packed columns, we have to use column
packing materials of small size, e.g. 3-5 p m particles. Since these packings are
retained in the column by still finer metal frits of e.g. OSpm, high-molecular
species tend to be filtered off on these frits. In addition, mechanical degrada-
tion of the polymers can take place in the frits or in the injection system, where,
to ensure a good injection quality, the injection pulse is transported at a very
high linear velocity through narrow-bore tubes before entering the region of
lower linear velocities in the wider-bore chromatographic column. With capil-
lary HDC there are no frits through which the polymers have to pass and the in-
jection can be made with good efficiency, without subjecting the polymers to
excessively high linear velocities. Yet, the absence of detectors (other than UV
absorbance) that can be used in narrow-bore capillaries is a serious drawback in
the capillary HDC technique.
4.4 HYDRODYNAMIC CHROMATOGRAPHY IN PACKED COLUMNS
The packing material in column HDC consists, in general, of spherical, im-

permeable spheres of e.g. glass, silica or ion-exchange resin. The separation of
differently sized species takes place in the interstitial volume between the parti-
cles, where due to a velocity profile with the lower velocities close to the parti-
cles, the larger species that are excluded from this region elute earlier than the
smaller species. Since the particles of the packing material generally are not uni-
form in size, the dimensions of the interstitial spaces between the particles is not
clearly defined. Also, the exact form of the velocity profile is not known. As a
result, estimation of the size of eluting species from relative residence times
alone, as can be done in microcapillary HDC, is hardly possible.
Ever since Small [l-31 pioneered this technique in the early 1970s, numerous
applications have been reported in the literature. Again, as in capillary HDC, the
majority of the articles deal with the separation of discrete particles. The popu-
larity of this technique stems, among other things, from the fact that under cer-
tain conditions, it is possible to construct a universal calibration curve. Nagy
[37] showed that latex particles with a different chemical composition, e.g.
polystyrene, vinyl acetate or vinyl chloride-ethylene, but also inorganic species
such as colloidal silica fall on the same curve of particle size versus relative
residence time. Thus, for particles of unknown composition, HDC in packed
columns makes it possible to calculate a reliable size. For a review of these
particle separations by packed column HDC, the reader is referred to McHugh
P11.
Again, applications of the separation of polymeric species are not numerous.
The majority are concerned with the characterization of xanthane polysaccharide
and polyacrylamide of high molecular mass, compounds that find use in en-
hanced oil recovery and as flocculants. Examples of the separation of these com-
pounds are given by Hoagland et al. [7] and Prudhomme et al. [6]. Since no
calibration standards are available, their results are expressed as the equivalent
particle diameter. A problem with the separations mentioned above is the gen-
erally poor separation between the polymer elution profile and the peak of the
low molecular mass component that is used to determine the t = 1 point, so often
the delayed marker injection technique, described by McGowan and Langhorst
[38], has to be used.
Langhorst and Stanley [39] used a low-angle light scattering detector to obtain
absolute molecular mass information on partially hydrolysed polyacrylamides.
Using a column filled with 15 pm ion-exchange particles, they found very broad
molecular mass distributions, extending up to a value of about 6 X lo7 Da.
These are values that cannot easily be determined by size exclusion chromatog-
raphy due to the lack of efficient, small-particle packing materials with large
enough pores. Lecourtier and Chauveteau [40] used a capillary model to predict
polymer velocity in flow through porous media. They found good agreement
between predictions and HDC experiments on xanthane in columns packed with
irregular silicon carbide particles.

122 Chapter 4
Other polymers that have been studied by packed-column HDC include dex-
tran [7], liposomes [41] and polystyrenes [42]. Mori et al. [5] used a column that
is a hybrid between a packed column and a microcapillary column by sintering
together a bundle of small-diameter glass rods. The resulting column, consisting
of numerous parallel microcolumns with hole sizes of 0.9-1.4 pm, was used in
HDC experiments on polystyrenes. Cheng [42] discussed the similarity and dif-
ference between size exclusion chromatography and packed-column HDC, using
separations of polystyrenes in columns packed with 16-20 and 1-4pm glass
beads.
The performance of column HDC can be expected to improve when very
small particles of a uniform size are used. Work in this field has been reported
by Kraak et al. [8] and Stegeman et al. [9] with separations of polystyrenes,
colloidal silica particles and proteins on impermeable silica spheres of 1.4-
2.67 p m diameter. To compare their experiments with microcapillary HDC, they
used the hydraulic radius, &, i.e. the radius of a capillary having the same sur-
face-to-volume ratio as the packed bed column, to characterize the average value
of the flow channel radius. From this they calculated the aspect ratios of polysty-
renes in these separations. & can be obtained as
(4.16)
where dpis the particle size and E is the bed porosity. In their experiments with
the 1.4 p m silica beads column, with a porosity of E = 0.380, the hydraulic radius
& amounts to only 0.286 pm, a size that, up till now, has never been attained for
a true microcapillary HDC column where the smallest column radius reported is
0.6pm [18]. Good separation of six polystyrenes of molecular mass 2.75 X lo6-
3.45 X lo4 Da and toluene in THF were shown in less than 6 min analysis time
(Fig. 4.15). They fitted their experimental relative residence times to the calcu-
lated aspect ratios for polystyrenes and obtained a relation in the form of Eq.
(4.7), so t = (1 + U,- C$)-, but with a value of C = 3.7 instead of C = 2.698
that would be expected for random coil macromolecules in a good solvent in a
cylindrical capillary. But as has already been pointed out in the theoretical sec-
tion, the exact value of C is not too important for t values in excess of say 0.95.
So with their columns, it is possible to determine the size of eluting species from
relative residence times alone, for polymer sizes up to 16nm in their column
packed with 1.4 p m particles (up to 30 nm diameter in the column packed with
2.67 p m material). This is important, as it indicates that a number of applications
that have been described in the section on microcapillary HDC can, in principle,
be performed on packed columns (although, at present, preparation of very uni-
form small particles and the efficient packing of these into a column is still a
problem). The main advantage of using a packed column is, of course, the fact
I . . ,
0 ' 2 4 6
time ,min
Fig. 4.15. HDC separation of polystyrenes and toluene in a column packed with 1.40pm diameter
non-porous silica spheres. Peaks 1-6, polystyrenes of molecular mass 2750, 1260, 700, 310, 127
and 34.5 ma,respectively; peak 7, toluene. Reprinted from [9].
that detectors other than the ultraviolet detectors used in microcapillary HDC
can be applied. Still, with the columns packed with these highly efficient, small
sized materials, not every standard liquid chromatographic detector can be used.
For instance, even a standard ultraviolet detector cannot be used as such, since a
detector cell volume of only a few microliters deteriorates the separation of the
very narrow eluting zones too much. Therefore, in [8,9], a small length of a
100pm internal diameter capillary was placed at the outlet of the HDC column
to act as the UV detector cell. Standard refractive index detectors cannot be used
either, but since this detection principle, using laser sources, has already been
described for use in capillary applications [43-45],this more or less universal
detection method could well be applied in HDC separations if the efficient, small
particle size packed columns were equipped with a capillary detection cell.
Another application of packed columns is the HDC separation that can take
place in the interstitial volume in a size exclusion chromatographic column [46].
Thus, there is a possibility of having a chromatographic column that can handle
Refirences pp. 125-126

124 Chapter 4
r I I I 1 I
0 10 20 30 40 50
t i m e , min
Fig. 4.16. HDC-SEC separation of polystyrenes and toluene on a column packed with 3 pm porous
silica particles (pore size 6 nm). Peaks 1-8, polystyrenes of molecular mass 4000, 2200, 775, 336,
127, 43.9, 12.5 and 2.2 kDa, respectively; peak 9, toluene. Reprinted from [47].
a very wide range of molecular masses, the smaller components being separated
in the pores of the packing material by a GPC mechanism, while the very large
molecules are separated by HDC. This has been studied by Stegeman et al. [47]
with GPC columns containing 3 ,urn porous silica particles ( 6 nm pore size) and
3 p m cross-linked poly(styrene-divinylbenzene) particles (10 and 30 nm pore
sizes). The separation of eight polystyrenes and toluene on the silica column by
this mixed mechanism is shown in Fig. 4.16. It is claimed that the separation
range of this column is for molecular masses ranging fiom a few hundred up to
2 X lo7 Da, a dynamic range that cannot be attained, without sacrificing selec-
tivity and efficiency, with columns working with a pure size exclusion mecha-
nism.
4.5 CONCLUSIONS
From this overview we have seen that HDC is used mainly for the separation
of discrete particles rather than for polymeric species. This technique, which
uses only simple instrumentation, is potentially very useful for the separation of
polymers, especially those with very high molecular masses. It has been shown
that both with columns packed with very small impermeable spheres of uniform
size, and with microcapillary columns, information on the size of separated spe-
cies can be obtained without recourse to calibration standards. The working
range is for molecular radii ranging from say 1 nm up to about 500 nm. Cur-
rently, the detection in microcapillary HDC is limited to UV absorption, but
promising new detection principles based on laser technology are emerging.
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E.R. Adlard @id.),Chromatography in the Petroleum Industry
Q 1995 Elsevier Science B.V. All rights reserved 127
CHAPTER 5
Chromatography in petroleum
geochemistry
S.J. Rowlanda and A.T. R e v i l l b

apeboleurn and Environmental Geochemistry Group, Department of Environmental Sciences, University of
Plymouth, Drake Circus, Plymouth, PL4 8AA, UK; bCSIRO Division of Oceanography, Castray Esplanade,
Hobart9Tasmania, Australia
5.1 INTRODUCTION
Petroleum is formed by the natural degradation of a heterogeneous macro-

molecular substance known as kerogen. Whilst a proportion of kerogen can be
made amenable to examination by chromatographic techniques if it is first de-
graded to more volatile compounds, it is for the analysis of petroleum itself that
chromatography has found most widespread use. Indeed, the complex nature of
petroleum makes the use of chromatographic methods more or less essential for
any detailed geochemical study. Pioneering studies, prior to the advent of mod-
ern day chromatography, employed distillation as the principal separation
method (see e.g. [13) but whilst distillation remains valuable, the large scale of
apparatus and samples required, the expertise necessary and the time-consuming
nature of the work have made chromatography-based methods much more
popular for geochemistry in recent times. This chapter reviews some of the most
recent developments.
5.1.1 Recent reviews
The use of chromatographic and other methods to characterize kerogen has

been reviewed by Rullkotter and Michaelis [2], whilst the applications of high
temperature gas chromatography (GC) to geochemistry have also been discussed
recently [3]. Horsfield [4]has reviewed the applications of pyrolysis techniques,

128 Chapter 5
such as pyrolysis-GC, to petroleum geochemistry and reviews published regu-

larly in Analytical Chemistry include useful summaries of some of the applica-
tions of chromatography in this field up to about 1991 (see e.g. [5,6]). Several
up-to-date texts have appeared which discuss the analysis of so-called bio-
logical marker compounds in petroleum [7-91. These compounds are valuable
for oil-source rock and oil-oil correlations and numerous applications of chroma-
tography in this area have been reported. Indeed, some techniques, such as GC
and GC-MS are used so frequently in this area that in the present review we have
selected only examples which are intended to illustrate particular advances.
Other chromatographic methods are used more rarely but represent areas of great
potential for future geochemical research and these are also discussed. Thus, we
have attempted a somewhat selective review, confining ourselves mainly to re-
cent (i.e. post- 1990) research.
5.2 KEROGEN AND OTHER PETROLEUM MACROMOLECULES
By definition, kerogen is insoluble in organic solvents [2] and is not amenable

to conventional chromatographic fractionation. Many studies have used bulk
characterization techniques but these have provided relatively little molecular
information. However, chromatography has played an increasingly important
role, usually in conjunction with degradation techniques such as thermal or se-
lective and non-selective chemical degradation. The latter reduce the macro-
molecular kerogen to smaller sub-units which are more amenable to chroma-
tographic analysis. Oxidative degradation usually produces functionalized com-
pounds such as acids which require derivatization prior to GC or GC-MS
[10,111. More selective chemical degradations have included sequential use of
desulphurizing agents such as nickel(0)cene and Li/EtNH2 for the characteriza-
tion of sulphur-rich kerogens and other macromolecular fractions of oils fol-
lowed by LC, TLC and GC [ 12,131. The more severe thermal techniques, such as
flash pyrolysis (py) are often interfaced directly to a chromatography system, for
which GC and GC-MS are most commonly used. Such techniques require small
samples, have a fast sample throughput and reduce the need for sample work-up.
A significant advance in this technique was the quantification of pyrolysis prod-
ucts via the addition of internal standards [ 141. The latter approach is illustrated
in Fig. 5.1 which shows a py-GC-MS total ion current chromatogram of a sample
of asphaltenes from an Iranian bunker oil admixed with poly-t-butylstyrene in-
ternal standard. Pyrolysis produces the t-butylstyrene monomer which can be
used to semi-quantify the pyrolysis products of the asphaltenes.
Py-GC methods have found increasing use in the studies of kerogen or other
macromolecules [ 15,161. For example in the determination of sulphur-rich kero-
Chromatography in petroleum geochemistry 129
Internal standard
100
90
TIC /
80
70
60
50
n-lcosane
40
30
YaneI I I J n-Triacon tane
20
10
0
Scan 500 1000 1500
R.T. 26: 2 1 52: 46 79: 11
Fig. 5.1. Pyrolysis-gas chromatography-mass spectrometry total ion current chromatogram of as-
phaltenes (ca. 200 pg) isolated from Iranian bunker oil mixed with t-butylpolystyrene internal
standard (10 pg) and pyrolysed at 600C for 20 s. Components from methane to triacontane can be
measured if the p+olysis behaviour of the internal standard is reproducible and linear. Kratos MS
25 GC-MS operated with CDS 120 pyroprobe (modified). GC oven temperature 4 0 to +300"C at
5C /min. GC column: 30 m X 0.32 mm DB-1 (J & W).
gens, unique distributions of pyrolysis compounds have provided prima facie

evidence that unsaturated kerogen moieties derived from algae take up sulphur
and form di- and polysulphide linkages [ 17,181. In other studies, py-GC-MS has
aided identification of the origins of polycyclic oligoterpenoids in crude oils via
resinous polymers [19] and has established that some kerogens are formed by
selective preservation of algaenan-containing thin outer walls of Chlorophyceae
algae [20].
The availability of a range of detectors is also important for the progress of
py-GC characterization of macromolecular geochemical materials. The most
common detector for GC is the flame ionization detector (FID), which is non-
selective and responds to almost any compound containing C-H bonds. The

130 Chapter 5
main use of py-GC-FID is the typing of kerogens [ 141 on the basis of the dis-
tribution of pyrolysate compounds and their relative quantities. However, ele-
ment specific detectors also have important applications in py-GC of kerogens.
Perhaps the most common of these is the flame photometric detector (FPD)
which is selective for sulphur-containing compounds. The sensitivity of both the
FID and FPD allows the utilization of column splitting technology to obtain con-
current chromatograms from both detectors [ 181.
A less widely used selective detector is the nitrogen-phosphorus detector
(NPD), generally used for nitrogen containing compounds. This has enabled
pyridines, quinolines and benzoquinolines to be observed in the pyrolysates of
asphaltenes, coals and source rocks [21].
The atomic emission detector (AED), which is a multi-element detector, has
also found increasing use and has typical limits of detection in the pg/s range for
several elements [22]. For example, alkylpyrroles have been detected recently in
kerogen pyrolysates by py-GC with FID-AED and MS detection [24]. AED al-
lows monitoring of several elements at once and importantly these include car-
bon, sulphur, nitrogen and oxygen, making it potentially a valuable tool in kero-
gen analysis [23]. An advantage of the technique is that the carbon chroma-
togram is very similar to that from an FID, making the two readily comparable.
In addition, the emission spectrum of a compound can be obtained to ensure it
contains the element in question. A drawback is that response is directly related
to the strength of the emission line which for some elements, e.g. oxygen, is very
weak.
One impressive multidetection study involved py-GC coupled with FTIR and
FID [25]. Fifty-nine pyrolysate compounds were determined in a peat deposit,
indicating a variety of decomposition reactions in the woody tissues. The use of
integrated chromatography detectors in similar studies may prove valuable in
future kerogen research.
5.3 GEOCHEMISTRY OF PETROLEUM
The degradation of kerogen which, as described above, is utilized by geo-

chemists to produce compounds which are easier to identify, is also accom-
plished by nature over much greater time spans and generally speaking, less se-
vere conditions. The product of this natural degradation or catagenesis is liquid
petroleum which is somewhat simpler to characterize by existing techniques than
the precursor macromolecules and a wide variety of chromatographic methods
have assisted in this characterization. Since fresh petroleum comprises compo-
nents which range from simple hydrocarbons to bulky micelles, a significant
proportion can be lost by evaporation if care is not taken in sampling and analy-
sis (e.g. use of special cylinders, such as Welker bombs, where the sample is
kept at its original high pressure) or pressurized oil syringe sampling methods
[26,27]. For this reason, most geochemical analyses are conducted on a distillate
fraction (so called CIS+ fraction) of petroleum where the volatile components
have been removed in a controlled manner. However, this may imply a signifi-
cant loss of information and whole oil analysis is usually to be preferred if at
all possible [28].
5.3.1 LC, TLC and TLC-FID
The instrumentation and equipment available for column LC and planar

chromatography has been reviewed [6]. Column LC with silica and/or alumina
stationary phases is a standard technique for petroleum fractionation, with or
without assisted gas pressure, and typically results in the isolation of so-called
saturate, aromatic and polar compound classes [7]. If care is taken to
avoid losses during subsequent evaporation of solvents from these fractions (e.g.
by use of Kuderna-Danish apparatus [29]), the resulting data can be reasonably
quantitative, particularly if internal standards are used to calibrate the measure-
ments [7,30]. However, some losses are unavoidable and recovery from the col-
umn is seldom 100%. Of the classes eluted, most detailed studies have continued
to concentrate on the examination of the saturate and aromatic hydrocarbon
fractions. However, a notable expansion of research into the heavy ends of
crudes (e.g. asphalt, tar and pitch fractions) has seen quite widespread applica-
tion of LC, for instance for the preparative isolation of acid-base fiactions of
asphaltenes and maltenes [3 11 and for characterization of asphaltenes before and
after visbreaking [32].
LC has also proved extremely useful for the isolation of compounds from
complex geochemical extracts of oils, coals and shales for structure elucidation.
The use of LC with silica gel and alumina columns and a variety of solvent
mixtures, followed by HPLC, allowed the isolation of seven alkylporphyrins and
thirteen porphyrin carboxylic acids from Messel shale [33], whilst pure vanadyl
porphyrins were isolated from other lacustrine sediments by LC on alumina fol-
lowed by propanesulphonic acid-bonded silica and HPLC [34]. Others have iso-
lated aromatic compounds such as basic nitrogen compounds (azaarenes) on sil-
ica [35] and 17 synthetic trimethylbiphenyls on silica and alumina [36]. Novel
C-ring cleaved triterpenoid aromatics were identified in Tertiary brown coal af-
ter LC fractionation of extracts on deactivated alumina and silica [37].
TLC is a cheap and rapid technique (especially if automated) which has found
widespread application for oil fractionation. However, losses of volatiles are
usually significant.

132 Chapter 5
Yamamoto [38] used TLC on silica gel to obtain 11 fractions of azaarenes

from crude oils and rock extracts from Japan and Sumatra.
Reversed phases (RP), impregnated-phases (e.g. Ag+) and multi-dimensional
TLC methods have not found very widespread use in oil geochemistry, unlike
many other areas 161 and future investigations may prove valuable for charac-
terization of polar fractions. For example, argentation TLC was useful in the
separation of various classes of sulphur-containing components from the malte-
nes of an immature bituminous shale [ l s ] and for the separation of unsaturated
components from the off-line pyrolysates of algal ultralaminae [20].
Non-planar TLC-FID (Iatroscan) is an under-utilized method which allows re-
producible (and if &+ fractions are examined, quantitative) analysis of whole
oils or oil fractions [39,40]. The method is particularly useful for replicate analy-
ses from which a meaningful statistical appraisal of results can be obtained. In-
teresting refinements include the use of Age-TLC-FID chromarods [41] and the
addition of a flame thermionic detector for the analysis of nitrogenous com-
pounds [42].
5.3.2 HPLC
HPLC is one of the most widely used modern chromatographic techniques

and numerous routine applications have been found in petroleum geochemistry.
Recent interesting studies include preparative and automated compound class
separations on silica and cyano-bonded silica with good (1 00 f 1.3%) recoveries
[43]; HPLC coupled with Iatroscan has been used for the quantitative analysis of
coal-derived oils [44] and determination of aromatics in oil distillates by 2-D
HPLC with column switching [45]. Use of reversed phases allowed Garrigues
and colleagues to study the retention behaviour of all 25 dimethylphenanthrene
isomers and to identify the compounds in crude oil [46] whilst a class separation
of nitrogen aromatics was possible with polar aminocyano-silica HPLC columns
[47]. Use of HPLC with refractive index detection facilitated identification of a
new series of rearranged hopanes in sediments and petroleum [48].
A quantitative method using backflush HPLC techniques allowed a determi-
nation of the heavy constituents of a crude oil to be made [49]. HPLC continues
to be routinely used for characterization of both free-base and metalloporphyrins
in oils and sediments [50] and new compounds are still being found [33,34]. In-
deed, HPLC coupled to element-selective detection is increasing in popularity
for the determination of metal speciation in oils (for review, see [51]). A recent
example is the determination of metalloporphyrins in coal extracts by HPLC-
ICP-MS [52] as illustrated in Fig. 5.2, showing the distribution of gallium por-
phyrins in coal. A conventional HPLC-UV chromatogram is shown for compari-
750
1 A
B
600
0
300
150
I I I I I
10 30 50 70 90 110
Time (min)
Fig. 5.2. HPLC-ICP-MS mass chromatogram (mlz 71) illustrating the distribution of gallium-
containing porphyrins in a coal from the British Coal National Coal Bank (Markham Main). Inset
shows more conventional HPLC-UV chromatogram. Mobile phase 1 mYmin, 1 mM t-butyl-
ammonium dihydrogen phosphate (150/) in methanol; column 30 mm X 3.9 mm octadecyl silane,
4p m Novapak (Waters). ICP-MS was VG Plasmaquad 2. For operating conditions, see [52].
son. Such methods show promise for the future study of metalloporphyrins, al-
though further research will be needed to achieve reproducible quantitation [52].
5.3.3 GC
Capillary GC continues to be used extensively for crude oil characterization

and can be considered a routine technique in the field, particularly for biomark-
ers and other hydrocarbons [7-91. A review of up-to-date GC publications has
appeared [ 5 ] . A recent study reported separation and identification of 21 light

134 Chapter 5
end, 83 whole oil and 17 heavy end components of Vlaska oil from the Adriatic
Basin [53].
Routine analyses typically use polydimethylsiloxane or phenylmethylsiloxane
stationary phases (e.g. OV-1 or OV-5 type) but even with these phases retention
indices of compounds are unfortunately reported irregularly. More unusual GC
phases are sometimes employed. For example, certain liquid crystal phases have
proved to be unrivalled for separation of isomers of alkylaromatics such as di-
methylphenanthrenes and dibenzothiophenes [46,54] although column availabil-
ity and the limited range of operating temperatures may place practical restric-
tions on their extensive use for crude oil analysis. Impressive separations can
also be obtained by 2-D GC; a good example is the quantitation of aa and &?
isomers of dimethylnaphthalenes for maturity index calculations with the added
advantage that no sample preseparation was needed [55]. Many of the compo-
nents of crude oil contain chiral centres and this stereochemistry confers high
information content on such molecules [7,9]. The separation of enantiomers of
various biomarkers on chiral GC phases is therefore of interest [56] and the col-
umns suitable for such analyses are readily available as a result of a dramatic
expansion of research into the GC properties of cyclodextrin phases.
An automated method for molecular weight determination of crude oils by GC
has been published [57] and a good correlation with the results of cryoscopic
molecular weights found. The usual molecular weight range of GC analyses (viz.
typically C1-C40)has also been considerably extended (e.g. to Cso) since capil-
lary high temperature GC columns have become commercially available (see
reviews [3,58-60] and Chapter 3 of this volume). Although SIMDIST GC analy-
ses have in fact allowed some separation of >C, components for some years
[61,62], present HTGC columns allow analysis of even relatively involatile or-
ganometallic species such as metalloporphyrins whilst retaining reasonable GC
resolution [63,64]. Indeed, when coupled with element selective detectors (e.g.
MIP, ICP-MS [22,65]) HTGC offers a rapid fingerprinting method for or-
ganometallic compounds in oils. The use of element-selective detection with GC
has been reviewed [66].
Despite the high resolving power and extended working range of modern
capillary GC, a substantial proportion (3690%) of fresh and biodegraded crude
oils is still unresolved and chromatograms of crude oils often contain humps
or so-called unresolved complex mixtures (UCMs). This is illustrated by the
flow diagram shown in Fig. 5.3 which shows that for heavily biodegraded Vene-
zuelan Tia Juana Pesado crude, which is imported into the UK for lubricant
manufacture; current methods result in a detailed knowledge of only about 5% of
the petroleum.
A few attempts to characterize hydrocarbon UCMs have been made recently
[67-703. These have used degradative oxidation followed by GC and GC-MS in
Asphaltene
-2 100%
(20% of oil) (80% of 011)
Aliphatic Aromatic
(23.2%of oil) (35.2% of oil) (21.6% of oil)
I I I I 1 I1 I
Re;ol;d
(2.3% of oil)
Unrgewlved
(20.9% of oil)
d
R
;;e
( I 8% of oil)
Unrgee3;ved
(33.4% of oil)
Fig. 5.3. The limitations of chromatography in petroleum geochemistry? Flow diagram illustrating
the lack of molecular information available for a biodegraded Venezuelan crude oil. Although a
greater proportion of non-biodegraded oils is identifiable, a substantial proportion of even these
remains as unresolved complex mixtures.
conjunction with other methods. The method typically produces about 20% of
resolvable and identifiable oxidized products, mainly acids, lactones and ketones
(Fig. 5.4). The bulk of the UCM, although oxidized, is still unresolved and re-
mains a challenge for chromatographers.
5.3.4 GC-MS
Capillary GC-MS has perhaps made the single greatest contribution to the
analysis of petroleum hydrocarbons in the last 20years and hundreds, if not
thousands, of compounds have been identified. For example, over 700 com-
pounds were identified in a shale oil [71] and 450 compounds in a coal tar an-
thracene oil [72] and coal tar pitch on two stationary phases [73]. However, it
should be emphasized, that the proportion of oil represented by these resolved
components is not always high, and many remain unresolved and unidentified. A
review lists retention indices for 400 terpenoids on methylsilicone and Carbowax
20M phases [74] and many of those compounds discovered in petroleum which
Refirences pp. 139-141

136 Chapter 5
4 4 12 20 20 36 44 52
0 .
L 0
1
12
20 25 36 52
4
Time (mid
Fig. 5.4. Gas chromatograms of (a) a fuel oil unresolved complex mixture of hydrocarbons and (b)
enhanced resolution achieved by oxidative degradation to n-acids (e), lactones and ketones. Re-
solved components comprise less than 1% in the fuel oil and about 20% in the oxidized oil
(reproduced from [70]).
provide petroleum geochemists with palaeoenvironment, maturity, biodegrada-

tion and migration information are terpenoids, so such data may be important.
The term biomarker has been used to describe those compounds whose struc-
tures suggest an unambiguous link with a precursor biochemical and such com-
pounds are used for oil-source rock and oil-oil correlations [7-91. Despite the
usefulness of MS and tandem MS in identifying many of these compounds, the
occurrence of many structural and stereoisomers of hydrocarbons in oils means
that GC retention time is still an extremely valuable guide to compound identity.
For example, an extended injection temperature hold time GC method was
needed to separate 14a(H), 17a(H) and 14&H), 17P(H) isomers of C21-norchol-
estanes in mature sediments [76]. A review of the compound classes discovered
is outside the scope of this chapter and readers are referred to other sources for a
fuller discussion [7-9,79,80] but recent advances have been made in the identifi-
cation of non-hydrocarbons such as thiophenes and sulphides [ 18,s13, phenols
[82] and naphthenic acids [83]. A rapid method of biomarker analysis in oils and
source rocks is thermal extraction GC-MS [841.
5.3.5 GC-isotope ratio-MS
GC-isotope ratio-mass spectrometry is a potentially valuable addition to the

petroleum geochemists armoury of instrumental techniques and commercial in-
struments are now available. The method allows the rapid determination of the
13CP2C ratio of individual organic compounds in complex mixtures, which is
useful for correlation and possibly for assignment of the biological origins of the
compounds [SS-881. However, GC co-elution of certain components may mean
that further developments in chromatography are necessary before accurate 613C
values can be obtained. For example, y-Cerane from eukaryotic heterotrophs was
found to co-elute with 3p (CH,) hopane from methanotrophs in extracts of the
Green River oil shale. Peak deconvolution was needed to obtain adequate 6I3C
data [89].
5.3.6 LC-MS
LC-MS has still not found wide application in petroleum geochemistry, even
though the combination of HPLC technology with mass spectrometry would ap-
pear to be a powerful combination for the characterization of petroleum frac-
tions. An efficient and innovative data reduction process has been developed for
LC-MS analysis of hydrocarbon mixtures [90] and LC-MS has been evaluated
for the analysis of lubricating oils [91] the hydrocarbons of which are predomi-

138 Chapter 5
nately UCMs [69,70]. LC-MS has proved to be extremely valuable for the
analysis of chlorophyll-derived pigments in sediments [92] and may therefore be
valuable for the examination of geological analogues of these pigments (viz.
geoporphyrins) in the future.
5.3.7 SFC, GPC and SEC
Some of the fractions of petroleum which are less accessible to the geochem-
ist (e.g. Heavy Ends) and which are not amenable to examination by GC and
GC-MS, can nonetheless be examined by SFC-GPC andor SEC methods. These
chromatographic techniques can be coupled with a variety of detectors including
FID and MS [93,94,97,98]. For instance, saturates, aromatics, resins and asphal-
tenes in petroleum were separated in 15 min by SFC-FID [94] and carbon num-
ber ranges up to C, can be eluted [95]. SFC-MS is well suited to PAH analysis
in fossil fuels [96] and has good selectivity for porphyrins [93].
Applications of GPC to petroleum and coal analysis have been reviewed [6].
GPC has been used to study the behaviour of C3G52petroporphyrins from Ath-
abasca asphaltenes [99] and maltenes from Utah bitumens were found to have
average molecular weights of 300 to nearly 12 000 [IOO] by GPC. Porphyrins,
aromatics and alkenes can be determined by GPC in petroleum without sample
preparation [loll.
SEC on pure silicon carbide columns has been used to determine the molecu-
lar weight of V and Ni complexes in heavy ends of petroleum (see [6]) and the
use of SEC with element-specific detectors has been reviewed [102]. The mo-
lecular weight distributions of aromatics and alkanes has been determined by
SEC and used to differentiate natural gas condensates from petroleum [ 1031.
5.4 SUMMARY
A wide variety of chromatographic techniques has been used in petroleum

geochemical studies, but GC and GC-MS remain overwhelmingly most popular,
and these can be considered as routine methods. Whilst this has led to the ac-
quisition of a great deal of knowledge about a small proportion of the compo-
nents of petroleum, much remains to be learned about the geochemistry of even
the hydrocarbons, a substantial proportion of which are left unresolved by the
best chromatographic methods. Methods such as LC-MS and SFC may prove
increasingly useful for fractions such as resins, asphaltenes and polars, which
have previously proven difficult to characterize.
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E.R. Adlard (Ed.), Chromatography in the Petroleum Industv
CHAPTER 6
The 0-FID and its applications in

petroleum product analysis
A. Sironi and G.R. Verga

Fisons Instruments, Strada Rivoltana, 20090 Rodano (Milan), Italy
6.1 INTRODUCTION
Some 10 years ago Schneider et al. presented a gas chromatographic detection

technique for the selective determination of oxygen-containing compounds [I].
In this publication, a post-column system was described where two microreac-
tors combined in series allowed a conventional FID to be operated as an oxygen
specific detector: the 0-FID was born!
Briefly the system consisted of a high temperature cracking reactor to convert
any oxygen-containing compounds separated by a GC capillary column into car-
bon monoxide and an FID equipped with a micromethanizer for the catalytic hy-
drogenation of CO and its detection as methane. Hydrocarbons are not detected
because they are hlly cracked and reduced to the elemental states, i.e. carbon
(graphite) and hydrogen.
The 0-FID, originally described as a stand-alone GC device, was primarily
used for the determination of oxygenated anti-knock additives in modern fuels in
response to the growing demand for analytical procedures able to comply with
the requirements of quality control and process development in the petroleum
industry. The 0-FID method in fact satisfied the requirements for processing
gasoline samples with a single-column, direct injection, valveless gas chroma-
tographic procedure for quantifling the oxygenates.
The potential of the technique proposed by Schneider and his co-workers has
now been fully exploited and implemented. The first commercially available 0-
FID analyzer was presented at the Eighth International Symposium on Capillary
Chromatography held in Riva del Garda in 1987 [2].
Referencespp. 157-158
I44 Chapter 6
6.2 OXYGENATES AS COMPONENTS OF MOTOR GASOLINE
Today, it is widely recognized and accepted that energy and environmental

problems are closely related so that any possible solution to one must, of neces-
sity, take into account and meet the requirements of both. In this light, a major
target in the automotive field has become the need to achieve the highest energy
yield while reducing environmental impact to a minimum.
As far as energy is concerned, the supply of oil, which is in practice the most
important source of energy for the automotive industry, is often threatened by
the instability of the international situation and this may lead to very serious
consequences.
From the environmental viewpoint, it was a common belief some years ago
that air pollution due to car emissions in the main urban areas had attained its
maximum value and that traffic was not supposed to increase any further since it
had already reached a critical value for the existing road system.
A number of studies carried out during the 1980s have proved that this as-
sumption was totally incorrect since the traffic density remained almost steady
but the time period and the areas of highest concentration and density have dra-
matically increased with unavoidable adverse effects on the emission of CO,
lead, hydrocarbons (HC) and oxides of nitrogen (NO,). The primary objectives
of each country and particularly the US and Europe has been to combine the pos-
sibility of reducing the emission of these dangerous, toxic compounds in the at-
mosphere while at the same time decreasing oil consumption through the use of
reformulated gasolines and alternative fuels. In addition, there is also the increas-
ing use of catalytic converters fitted as standard in the exhaust systems of most
modern automobiles put on the market since the early 1980s in those European
countries where sensitivity to environmental problems was particularly high.
The accelerated phase-out timetable in the USA and Europe has imposed
major changes in fuel specifications. The need to maintain octane without the
use of lead compounds has forced refiners to look for alternatives, mainly aro-
matics such as benzene, toluene, and xylene (BTX) or oxygenated compounds
such as methyl tertiary butyl ether (MTBE), gasoline tertiary butyl alcohol
(GTBA), methanol and ethanol.
Oxygenated enhancers of either natural or industrial origin have therefore
found wider and wider application as fuel components because their premium
antiknock rating enables the preparation of unleaded gasolines with high octane
number suitable for high efficiency engines. Additionally their reactivity and
combustion properties provide a significant reduction of emitted pollutants with
evident advantages for the environment. Studies performed in the USA and
Europe have demonstrated that when used in gasoline, oxygenates greatly con-
tribute to the reduction of:
The 0-FID and its applications in petroleum product analysis 145
- CO emission by 25-30%
- benzene emission by about 15-20%
- particulate emission and associated PNAs by about 40%
- the reactivity of the emissions with consequent decreased probability of
ozone formation
- lead level in the environment with a remarkable decrease in blood concen-
trations following the lead phase-out program as shown in Fig. 6.1.
Finally, it is also important to emphasize that when oxygenated compounds
are used as neat fuels, they can easily replace gasoline in cars causing a marked
reduction of ozone and when used in place of diesel fuels they contribute to a
considerable reduction of particulate and NO, emissions.
The European Union, following a program of energy and environment con-
servation has also been moving closer and closer to gasoline with no lead. An
EU decree reduced the lead content from 0.4 g/1 to a limit of 0.15 g/1 from 1986
and it was projected that all member states had to have unleaded gasoline avail-
able by July 1989. Lead phase-out in Europe was completed in July 1992 and
several countries have followed the examples of Germany, France, and the UK
in introducing premium gasoline (98 RON) in the marketplace. At that date,
unleaded gasoline occupied 43% of Europes pool, but this average, in reality,
concealed a wide range from country to country. While German gasoline, for
example, consists of almost 80% unleaded and growing, fuels sold in Mediterra-
Toul lMd wed per B month period

(mllllon ton.)
110
100 11
90 14
80 1s
70 12
00 11
50 10
-
40
1977 . 1978 . 1979 .
-
1976 1980
Amrage Mood lead I m l a
Lead wed In gaeollne
Fig. 6.1. Correlation between the use of lead in petrol and blood lead level. (Source: USA Envi-
ronmental Protection Agency. Taken from EFOA Newsletter Issues No. 2, June 1988.)

146 Chapter 6
TABLE 6.1
LIMITS FOR ORGANIC OXYGENATE COMPOUNDS IN FUEL BLENDS ACCORDING TO
EEC COUNCIL DIRECTIVE NO. 85-536 OF 5 DECEMBER 1985
Oxygenates Aa Bb
(vol. Yo) (vol. %)
Methanol (stabilizers to be added) 3 3

Ethanol (stabilizers may be added) 5 5
Iso-propyl alcohol 5 10
TBA 7 7
Iso-butyl alcohol 7 10
Ethers, with five or more C-atoms 10 15
Other oxygenates 7 10
Total oxygen content by weight % 2.5 3.7
nean countries are still largely leaded although unleaded gasoline sales are now
increasing.
This encouraging result in Europe has also been made possible by the EEC
Council Directive of 5th December 1985 on crude oil savings through the use of
substitute fuel components in petrol. This directive stated the limits of oxygen-
ates valid throughout Europe for reformulated gasoline. Table 6.1 reports these
limits for the individual components and also shows the total oxygen content
permitted.
6.2.1 Determination of oxygenates in unleaded fuels
As stated above, unleaded gasoline has received increased worldwide atten-

tion with the natural consequence that the importance of oxygenated compounds
in the petroleum industry has also increased and is expected to further grow in
the future.
Similarly, the demand for routine analytical methods for quality control and
process development has also grown with the more widespread use of these oxy-
genated enhancers in fuels. An ad hoc European committee, CEN/TC 19
WG9, was charged with the evaluation of methods available for this analysis
which, at that time, included infrared [3] and near infrared [4]spectroscopy, nu-
clear magnetic resonance [5], liquid chromatography [6,7] and also sophisticated
GC techniques coupled with mass spectrometry [8,9] and with microwave-
induced plasma emission [ 10,l I]. Other GC methods have also been taken into
consideration including those based on multidimensional techniques using both
packed and capillary columns and switching valves [12-171.
In general, all these methods were able to provide sufficient sensitivity and
resolution for the major oxygenated compounds present in fuels but many lacked
the important capability of being able to detect all oxygenates individually as
requested by the CEN/TC 19 WG9 or they were too matrix-dependent or too
expensive or, finally, limited by operational complexity [ 18,191.
In this light and after a careful evaluation of the different options available,
CEN/TC 19 WG9 decided to adopt the 0-FID method which is based on the se-
lective gas chromatographic analysis of oxygen-containing compounds with a
modified flame ionization detector [ 1,2,20]. This method has been extensively
tested for a number of years through severe interlaboratory tests which have
proved the use of the 0-FID to be the best method in terms of accuracy and pre-
cision.
The 0-FID method has become the official European standard method for the
selective determination of individual oxygenates in gasoline and is recognized as
the only one valid in case of dispute.
6.3 0-FID ANALYSER
The 0-FID analyser is essentially a high resolution GC fitted with a split in-
jector and equipped with two microreactors between the separating capillary
column and the flame ionization detector. The cracking reactor, connected im-
mediately after the capillary column, converts oxygenated compounds into car-
bon monoxide, while the micromethanizer, inserted in the FID jet, converts the
CO into methane, which is then detected by the FID. Figure 6.2 shows the sche-
matic configuration of the 0-FID analyser.
6.3.1 Cracking reactor
The cracking reactor (Fig. 6.3) consists of a platinudrhodium capillary tube

coiled around a special insulating support. The cracking element, thermally and
electrically insulated, is directly heated by means of a low voltage power supply
and temperature controlled in the range 200-1200C. To protect the Pt/Rh ele-
ment against air oxidation, a stream of nitrogen flows into the cracking reactor
from a purge line maintaining the reactor in an atmosphere of inert gas. When an
oxygenated compound eluted from the capillary column enters the reactor,
heated at a temperature higher than 1000C, each atom of oxygen will produce a
molecule of CO while all hydrocarbons will be decomposed into carbon and hy-
drogen according to the following reactions:

148 Chapter 6
C,HmOx-+ x C 0 + %mH, + (n - x)C (6.1)
The carbon formed in both reactions deposits out as a graphite layer on the
internal walls of Pt/Rh tube, further promoting the conversion process.
Under the assumption of carbon in excess, the following reactions can take
place:
c + coz -+ 2 c o (6.3)
C + H,O -+ CO + H, (6.4)
It is possible to verify that at a working temperature of 1100-1 150C, the

equilibria of the reactions (6.3) and (6.4) are completely shifted to CO forma-
tion. A certain amount of carbon deposited on the reactor walls is continuously
required to maintain the reactions involved in the pyrolysis process under kinetic
control. For this reason, a continuous deposit of carbon within the Pt/Rh capil-
lary tube is obtained by doping the carrier gas (normally nitrogen) with a hydro-
carbon such as pentane or hexane at 10-50 ppm [2]. Alternatively a carbon layer
1 - FID
Hydrogenation
microreactor
Gas
filter
Carrier
Capillan/ temperature
column cracker
Fig. 6.2. Schematic diagram of the O-FID analytical system.
The 0-FIDand its applications in petroleum product analysis 149
+
Purge gas line
7
Temperature Power supply
sensor
- Cracking
element
Insulating -
material
Capillary
column
it
Fig. 6.3. Principle of the cracking reactor.
Transfer line
can be provided by the injection of several microliters of a hydrocarbon before

starting the analytical cycle [ 1,2 11.
The presence of oxygen due to leaks in the system or to other oxygenated im-
purities in the carrier gas depletes the carbon layer, reducing the cracking capac-
ity and the lifetime of the reactor. In addition approximately 1% of hydrogen has
to be mixed with the carrier gas to counteract chemisorption effects and promote
the pyrolysis process.
These requirements are met by connecting an auxiliary hydrogen line to the
carrier line before it enters the split injector.
The cracking capacity of the reactor, that defines the selectivity of the O-FID
system and its application range, is strictly dependent upon the reactor tempera-
ture and, to a minor extent, on the type of compound to be cracked. As reported
by Schneider et al. [I], the full conversion into carbon and hydrogen can range
between 2 and 20pg for aliphatic hydrocarbons (n-heptane) with a reactor tem-
perature between 1200 and 1400C (Fig. 6.4).Temperature increase, however,
drastically reduces the lifetime of the Pt/Rh cracking element. For this reason, a

150 Chapter 6
% conversion
-
14OOOC
70 I I I I 1 1 I I I , I I
0 10 20 30 40 I
nHeptane (pg)
Fig. 6.4. Cracking efficiency for n-heptane as a h c t i o n of reactor temperature. FID (without mi-
cromethanizer) temperature, 350C.
practical compromise must be reached between the expected selectivity (carbon

rejection) and the cracker lifetime. Under standard temperature condition (1 100-
1200C) and nitrogen purge flow, the lifetime of the reactor is expected to be in
the range of 1500-2000 h. After extensive operation or after high temperature
operation, the Pt/Rh tube can become brittle and porous, allowing oxygen (air) to
enter the tube and cause depletion of the carbon layer. Under these conditions,
the cracker cannot work properly and generally needs to be replaced as the re-
sulting drastic decrease in selectivity and sensitivity cannot be reversed.
6.3.I . I Low temperature cracker
Considering that cracking reactor lifetime is a vital point in the economy of
the 0-FID, efforts have been made to overcome this limitation. Initially im-
provement was obtained by using a purging line inside the PtlRh tube holder in
order to maintain the cracking element under nitrogen instead of air to avoid
oxide formation. More recently, a new cracking reactor able to operate at tem-
peratures lower than 850C has been developed. This has been achieved by
means of a new catalytic cracker manufactured by Fisons Instruments (patent
pending). This low temperature reactor extends the lifetime to several thousands
hours while preserving the same cracking capacity.
6.3.2 Hydrogenation microreactor
The catalytic hydrogenation reactor (Fig. 6.5) converts the carbon monoxide
produced by the cracking reactor into methane according to the well known re-
action
The micromethanizer consists of a short PLOT glass capillary tube coated

with aluminium oxide on which a nickel catalyst is adsorbed in metallic form.
jet
Ni catalyst
iealte r
+ H2
Air +=
Transfer line
Fig. 6.5. FID base body with catalytic hydrogenation microreactor (methanizer).

152 Chapter 6
This microreactor, directly inserted into the standard FID base body, is sup-
plied with the same hydrogen flow used for the detector and it is kept at an op-
erating temperature of 330-350C. The micromethanizer is connected to the
cracking reactor outlet by means of a short fused silica transfer line of virtually
zero dead volume.
6.4 ANALYTICAL PROCEDURE
The sample, generally with an added internal standard, is split injected and the
oxygen-containing compounds are separated by a suitable capillary column. A
prior separation of hydrocarbons or other carbon-based compounds in the sample
is not required. As a consequence, the analysis time is limited to the elution of
the last oxygenated compound. The analysis is usually performed isothermally,
which further reduces the total analysis time. The split injection system is used
as standard and the splitting ratio (1:100/200) as well as the injection volume
(0.5-1 pl) are selected to maintain the 0-FID within its cracking capacity. The
absolute amount of a sample that can be completely cracked represents the ca-
pacity of the cracking element which is dependent on the reactor temperature
and on the type of molecule to be cracked (e.g. aromatic hydrocarbons are
cracked easier than aliphatics).
As previously discussed, increasing the temperature of the reactor increases
the capacity but drastically reduces the lifetime of the PtRh reactor tube. By
maintaining the operating temperature at 85OoC, the cracker lifetime is extend-
ed while still preserving the required selectivity and sensitivity. Peak identifi-
cation is performed on the basis of retention times while quantitation is gener-
ally accomplished by means of an internal standard. In addition, it has to be
remembered that oxygen (the first peak eluted) and water are normally de-
tected.
6.4.1 Quantitative analysis
Considering that any oxygen atom is stoichiometrically converted into a

methane molecule, the 0-FID response for oxygenated compounds is strictly
related to their oxygen content and because of this, a preliminary calibration is
normally required for response factor determination. Calibration mixtures can be
prepared gravimetrically by blending the oxygen-containing compounds with
gasoline that has been previously found to be free of oxygenates or with a syn-
thetic mixture of hydrocarbons. A known amount of internal standard is added as
reference to reduce errors caused by variations in injection volumes.
The peak area of each oxygenate and of internal standard are determined and
the response factors relative to the internal standard can be calculated as follows:
where RF is the response factor, A, is the area of the internal standard peak, A, is
the area of the oxygenate peak, Qsis the quantity of internal standard added in
the calibration mixture, Qo is the quantity of the oxygenate in the calibration
mixture.
Alternatively, for any identified oxygenated compound, a theoretical factor
can be easily calculated in advance. The good correspondence between the ex-
perimentally determined factors and the calculated theoretical values allows the
calibration procedure to be simplified for one component only, generally the in-
ternal standard. Table 6.2 shows calculated response factors for the most com-
mon oxygenated compounds of interest using methyl propyl ketone (MPK) an n-
propanol (1-PROPOH) as reference standards. The chromatogram in Fig. 6.6
TABLE 6.2
CALCULATED RESPONSE FACTORS FOR THE MOST COMMON OXYGENATED COM-
POUNDS OF INTEREST IN GASOLINE ANALYSIS
Oxygenate Response factors
DEE, diethyl ether 1.233 0.860

DIPE, di-isopropyl ether 1.701 1.186
MTBE, methyl tert-butyl ether 1.467 1.023
ETBE, ethyl tert-butyl ether 1.701 1.186
BME, butyl methyl ether 1.467 1.023
DPE, dipropyl ether 1.701 1.186
TAME, tert-amyl methyl ether 1.701 1.186
Acetone 0.967 0.674
DIBE, di-isobutyl ether 2.167 1.510
MEK, methyl ethyl ketone 1.199 0.837
MEOH, methanol 0.533 0.372
tert-BUTOH, tert-butanol 1.233 0.860
2-PROPOH, 2-propanol 1 .oo 0.697
ETOH, ethanol 0.766 0.535
DBE, dibutyl ether 2.167 1.510
MPK, methyl propyl ketone 1.433 1 .OOoa
sec-BUTOH, sec-butanol 1.233 0.860
1 -PROPOH, n-propanol 1.000a 0.697
iso-BUTOH, isobutanol 1.233 0.860
1-BUTOH, n-butanol 1.233 0.860
acornpounds chosen as reference with response factor of 1 .OOO.

154 Chapter 6
NAME % MASS NAME Yo MASS

1) DlPE 1.49 10) Ten-ButOH 1.92
2) MTBE 0.76 11) 2-PropOH 2.68
3) ETBE I.49 12) DBE 0.76
4) BME 0.71 13) See-ButOH 1.14
5) DPE 1.13 14) 1-PropOH (IS.) 2.01
6) TAME 2.63 9 15) Water 0.04
7) Acetone 0.58 11 16) IsobutOH 2.69
8) DlBE 1.15 17) I-ButOH 1.13
9) MEOH 1.5
r
i
i
2.
12
5
L
0 5 10 15 20 min.
I I
35 35 80 Temperature ("C)
Fig. 6.6. Determination of oxygenates in unleaded gasoline. Column, 30 m X 0.25 mm id.; DB
WAX (J&W) 0.5 p m film thickness. Sample injected, 1 pI (split 1:200). Cracker temperature,
850C.
refers to the analysis of a gasoline sample where the quantitation has been per-
formed using n-propanol as internal standard and the calculated response factors
(Table 6.2)
6.4.2 Total oxygen determination
The primary purpose of the 0-FID is to determine the content of oxygenated

compounds in a complex hydrocarbon matrix. The total mass percent of the or-
ganic bonded oxygen in the sample may also be determined without the need to
identify any individual oxygenate and without complete resolution. The total
1) Oxygen
2) Methyl isopropyl ether
3) Methyl-tert-butyl ether
4 4) Diisopropyl ether
5) Methyl-sec-butyl ether
6) Ethyl-tert-butyl ether
0-FI D 7) Methyl-nbutyl ether
8) Ethyl-isobutyl ether
9) Isopropyl-tert-butylether
10) Dipropyl ether
11) Ethyl-nbutyl ether
12) Methyl-npentyl ether
13) sec-Butyl-tert-butylether
f ' " ' 1 . ' . . I . . ' . I . " ' I .

6 11 16 21 26 min.
Fig. 6.7. Determination of ethers (0.2% each) in gasoline sample. Comparison of the 0-FID re-
sponse with the FID trace. Column, 97 m X 0.2 mm i.d. UCON 550 LBX, 0.2pm film thickness.
Column temperature, 60C.
oxygen content is easily calculated by summation of all the peak areas, exclud-
ing dissolved oxygen, water and internal standard.
6.4.2.I Selectivity for oxygenates and sensitivity
The 0-FID method is highly selective, allowing the non-oxygenated com-
pounds (e.g. hydrocarbons) to be entirely suppressed when the cracker operates
within its capacity limits and the selectivity (oxygenhydrocarbon) approaches
lo6. To give an idea of the selectivity, the 0-FID and standard FID chroma-
tograms of a sample of gasoline spiked with ethers are shown in Fig. 6.7.
As shown in Fig. 6.8 with a blend containing 200 ppm by weight of each oxy-
genate, the 0-FID response is strictly related to the oxygen content of the mole-
cule entering the cracker. It is also clear that the sensitivity is lower in compari-
Referencespp. 15 7-1 58
156 Chapter 6
1) Oxygen
2) Acetone
3) Ethyl acetate
4) Methyl alcohol
5) lsopropyl alcohol
6) Ethyl alcohol
7) sec-Butyl alcohol
2 6 8) 1-Propano1
9) Water
I I I I
0 4 8 10 min.
Fig. 6.8. Detection of oxygenates in gasoline at 200 ppm level. Column, 30 m X 0.25 mm i.d. DB
WAX (J&W) 0.5pm film thickness. Column temperature, 50C. Sample injected, lyl (split
1 :100). Cracker temperature, 850C.
son to a standard FID since any oxygenated compound is reduced to methane

and the FID response is mass-dependent.
The detection limit is, therefore, strictly related to the expected selectivity,
sample amount entering the column, column resolution and matrix composition.
The O-FID was developed specifically for the selective determination of in-
dividual oxygenates in fuels. For this purpose, it represents an ideal tool comply-
ing with the sensitivity requirements of official methods (0.1-20% by mass). A
detection limit of 20-30 ppm as methanol can be estimated by injecting 1 p l of
gasoline with a split ratio of 1:lOO. The detector is linear over a range of about
1 O5 which is lower than a standard FID.
6.5 O-FID APPLICATIONS
Besides the determination of oxygenates in gasoline, the O-FID can be used

for any other GC applications where oxygen-containing compounds have to be
selectivity detected in complex organic matrices. For example, the use of the 0-
FID has been reported for the determination of oxygenated species in coal-
derived liquid fuels which have been recently investigated as alternative feed-
stocks [22].
Since the 0-FID response is proportional to the oxygen content, compounds
containing several oxygen atoms can be analysed with evident advantage. This is
the case in the GC determination of some glycol ethers and glycol ether acetates
in biological matrices [23].
Other applications have been described such as the characterization of fla-
vours and fragrances used in the cosmetic industry [24] and the investigation of
essential oils of pharmaceutical importance [25]. The use of the 0-FID for these
and other applications will no doubt increase in the future.
6.6 CONCLUSION
The 0-FID had been widely accepted as a reliable method for oxygenates de-
termination in fuels. The main features can be summarized as follows:
(1) simple to operate, single column, valveless direct injection GC technique;
(2) highly selective response to oxygenates, totally uninfluenced by sample
matrix;
(3) total oxygen determination capability, not related to the identification of
individual oxygenates;
(4) short analysis time, limited only by the elution of oxygenated compounds;
(5) simple calibration procedure.
Weak points of the 0-FID are the relatively limited cracking capacity and the
low sensitivity and linearity compared to a standard FID. Future possible 0-FID
developments may overcome these limitations and extend the range of analytical
interest.
6.7 REFERENCES
1 W. Schneider, J.C. Frohne, and H. Bruderreck, J. Chromatogr. 245 (1982) 71.

2 G.R. Verga, A. Sironi, W. Schneider and J.C. Frohne, High Resolut. Chromatogr. Chroma-
togr. Commun. 1 1 (1988) 248.
3 D.R. Battiste, S.E. Fry, F.T. White, M.W. Scoggins and T.B. Mc Williams, Anal. Chem. 53
(1981) 1096.
4 J.L. Wong and B. Jaselskis, Analyst 107 (1982) 1282.
5 G.E. Renzoni, E.G. Shankland, J.A. Gaines and J.B. Callis, Anal. Chem. 57 (1985) 2864.
6 M. Zinbo, Anal. Chem. 56 (1984) 244.
7 R.E. Pauls, J. Chromatogr. Sci. 23 (1985) 437.
8 R. Orlando and B. Munson, Anal. Chem. 58 (1986) 2788.
158 Chapter 6
9 J.H. Shofstahl and J.K. Hardy, Anal. Chem. 58 (1986) 2412.

10 K.J. Slatkavitz, P.C. Uden and R.M. Barnes, J. Chromatogr. 355 (1986) 117.
11 C. Bradley and J.W. Carnahan, Anal. Chem. 60 (1988) 858.
12 J. Sevcik, High Resolut. Chromatogr. Chromatogr. Commun. 3 (1980) 166.
13 R.E. P a d s and R.W. McCoy, J. Chromatogr. Sci. 19 (1981) 558.
14 A.F. Lockwood and B.D. Caddock, Chromatographia 17 (1983) 65.
15 L.A. Luke and J.E. Ray, Analyst 109 (1984) 989.
16 J.M. Levy and J.A. Yancey, High Resolut. Chromatogr. Chromatogr. Commun. 9 (1986) 383.
17 G.V. Johnson, J. Chromatogr. Sci. 25 (1987) 65.
18 N.G. Johansen, High Resolut. Chromatogr. Chromatogr. Commun. 7 (1984) 487.
19 S.W.S. McCreadie, D.F. Swan, G.M. Ogle and R. Pintus, Proc. 6th Int. Symp. on Capillary
Chromatography, Riva del Garda, Italy, 1985. Huethig, Heidelberg (1985), p. 456.
20 D. Steinmueller, International Lab. June (1988).
21 F.P. Di Sanzo, J. Chromatogr. Sci. 28 (1990) 73.
22 R.B. Howard, E.M. Steward, S.D. Anderson and J.L. Moler, Pittsburgh Conf., New York
March (1 990), Paper 1107.
23 C. Giachetti, G.R. Verga, G. Zanolo and F. Perovanni, unpublished.
24 K. Lakszner and L. Szepesy, Chromatographia, 26 (1988) 91.
25 K. Lakszner, L. Szepesy, I. Torok and E. Csapo-Barthos, Chromatographia 30 (1990) 47.
Journal of Chromatography Library Series, Val. 56
0 1995 Elsevier Science B.V.All rights reserved 159
CHAPTER 7
Microwave plasma detectors
Arie de Wit and Jan Beens

KoninklijkeShell-Laboratorium,Amsterdam (Shell Research B. V.), Badhuisweg 3,
1031 CMAmsterdam, The Nethedands
7.1 INTRODUCTION
Detectors in gas chromatography (GC) can be roughly divided into two cate-
gories:
1. Detectors that are virtually non-selective, responding to every component

in the carrier gas. Examples are the thermal conductivity and the ultra-
sonic detector.
2. Detectors that are selective, responding only to certain properties, e.g. UV
absorption, electron affinity. Examples are the flame photometric, Hall
conductivity and electron capture detectors.
In general, selective detectors are used to facilitate detection of a particular

compound in the presence of a high background signal due to insufficient sepa-
ration from other components. These selective detectors are also used to obtain
qualitative information on the presence of hetero atoms in the gas chroma-
tographic effluent. However, most of these detectors can only determine one
type of hetero atom, which implies that in a diversified laboratory, many differ-
ent selective detectors have to be used. For the determination of elemental ratios,
two or more selective detectors have to be operated simultaneously. Such a sys-
tem is very complicated and sometimes it is difficult to combine two or more
selective detectors. In addition, some detectors respond to a class of compounds,
e.g. the electron capture detector responds to all halogens and not just to chlorine
or bromine. Furthermore, no selective detectors are available for hydrogen, oxy-
gen and the noble gases. A detector that is capable of monitoring any of a large
Referencesp. 200
160 Chapter 7
number of elements with a high degree of selectivity and sensitivity would there-
fore be highly desirable.
Ideally, a selective, multi-element detection system for gas chromatography
should:
1 . measure simultaneously all elements which can emerge from a gas chroma-
tographic column;
2. have a very low limit of detection for each element, at least better than 100
Pg/%
3 . have a wide linear dynamic range for each element, e.g. from 100 pg/s up
to at least 10 pg/s;
4. have a fast response, to enable the determination of components eluting
from narrow-bore capillary columns;
5 . insensitive towards all other elements, i.e. the selectivity ratio should be
be
better than 10 000: 1;
6. yield a response independent of molecular structure.
Spectrometric detection systems based on measurement of atomic weight and

of atomic emission can potentially fulfil these requirements. Mass spectrometry
can measure either the mass of a molecule or the mass of the fragments pro-
duced. The molecular mass does not produce the desired information and from
the mass of the fragments, it is difficult to obtain the elemental information, es-
pecially when large amounts of other fragments are present. In atomic emission
spectroscopy, each atom produces its own pattern of emission lines. As every
atom can produce an emission line, in principle each atom can selectively be
determined.
7.2 PRINCIPLE OF OPERATION OF AN ATOMIC EMISSION

DETECTOR (AED)
The principle of an atomic emission detector is based on the spectral atomic

emission which occurs when a gas is heated to temperatures above a few thou-
sand degrees centigrade. This gas constitutes a distinct fourth state of matter -
the plasma - which differs from ordinary gases particularly as regards the follow-
ing properties.
Apart from the collisions occurring in a normal gas between individual mole-
cules, as a result of the applied energy, there is a considerable number of colli-
sions that lead to the production of atoms in excited quantum states. These proc-
esses involve the formation of various particles: molecules, atoms, positive and
negative ions, electrons, photons, etc. Because of the presence of electrons and
Microwave plasma detectors 161
ions, electric and magnetic fields can act upon this ionized gas. Being conduc-
tive, the gas mixture can absorb energy from an electric or magnetic field or
transfer energy to it. The mixture as a whole is neutral, because at every point
the concentrations of positive and negative charges roughly equal each other.
The state of a plasma which is enclosed in a chamber with walls having the
same temperature as the plasma can be described by a few parameters: tempera-
ture, mass density, energy density and composition. If the walls of the chamber
are at a lower temperature than the plasma, energy is dissipated on account of
radiation and heat conduction. In order to obtain a stationary state, this energy
loss has to be compensated for by heating the plasma. This heating can be done
in a chemical way, by means of a flame, however the maximum temperature is
then only about 2000C. The most efficient way to obtain higher temperatures is
by using electrical energy.
The plasma can be generated within a quartz tube, provided with two elec-
trodes with an electric current forced between them. The fluorescent passed
lamp is a well-known example of such a plasma tube. Depending upon the type
of current, the plasma is called an A.C. or D.C. current plasma. The plasma can
also be generated in a quartz tube placed within an electromagnetic field. This
electromagnetic field can be created within a coil, induced by an electrical cur-
rent, and the plasma is then called Inductively Coupled Plasma. The most effi-
cient power transfer from generator to plasma occurs when the quartz tube is
placed within a resonance cavity. When the resonance frequency of the vessel is
the same as the frequency of the generator, a strong magnetic field is generated
within the cavity. The quartz tube is placed in a position where the electromag-
netic field strength is maximal. The generator has to replenish the loss of energy
in the cavity, by means of a small antenna in the cavity. For practical reasons the
diameter of the cavity is between 1 and 10 cm so the resonance frequency is in
the order of 1-5 GHz. Generators with sufficient power (50-500 W) and a fre-
quency of 2.45 GHz are generally used ( e g the magnetron oven in the kitchen).
Waves of these lengths are known as microwaves and the plasma generated is
called Microwave (induced) Plasma. As an illustration the power per unit vol-
ume transferred to the various types is given below.
Fluorescent tube 0.1 Wlcm3

DCIAC plasma 10 WIcm3
ICP 100 Wlcm3
Microwave 10000 WIcm3
The energy of excited atoms and molecules is liberated as radiation when the
electrons return to the ground state. They emit light of a varying number of very
discrete wavelengths. This light is collected by a mirror or lens and subsequently
References p . 200
162 Chapter 7
one or more of these emission lines are filtered by an optical filter or spectrome-
ter. The intensity of the line is a measure of the concentration of the selected
atom.
7.3 HISTORICAL DEVELOPMENT OF THE PLASMA DETECTOR
Broida and Morgan [ 13 were the first, in 1952, to describe a system for the
analysis of gaseous mixtures of hydrogen, deuterium and air based on optical
emission spectroscopy with photoelectric detection. An electrodeless discharge
at 150 MHz in a continuous flow gas system was used in conjunction with a
high-resolution grating monochromator. Each component could be determined
with an accuracy better than 0.1 % of the total mixture. Gas chromatography was
hardly in use at that time, and it was not until 1965 that McCormack, et al. [2]
first reported a system in which the effluent of a gas chromatographic column
was fed into an argon microwave discharge of 2450 MHz at atmospheric pres-
sure. The emission was detected in the W-visible region. Under the conditions
described by these authors the eluted components were only partly fragmented in
the plasma and the emitted spectrum consisted of both atomic and molecular
spectra. They found very low selectivity ratios relative to n-hexane (between 10
and loo), the sensitivity being strongly dependent on the compound involved.
Bache and Lisk [3-61 were able to detect selectively the atomic lines of halogens,
phosphorus and sulphur: they used a low-pressure microwave-induced plasma of
helium providing nearly complete atomization of organic compounds. However,
reproducible operation was impeded by the deposition of carbon on the wall of
the quartz tube in which the plasma was confined.
Moye [7] used an almost similar system for the detection of phosphorus and
halogen compounds. He used argon-helium gas mixtures and found lower limits
of detection, between 0.1 and 1 ng. He examined a few experimental parameters,
such as the quartz tube diameter. The GC column was operated under reduced
pressure and the pressure in the plasma was maintained at 30 mbar. Experimen-
tal work was also done with DC plasmas. In 1968 Braman and Dynako [8] used a
DC discharge in helium together with a GC column. These authors used interfer-
ence filters or a spectrometer to yield limits of detection in the picogram-per-
second range for elements such as F, C1, Br and I. The maximum amount of car-
bon that could enter the detector was 0.1 mg with an optimum power supplied to
the plasma of 10 W. Dagnall [9] used a microwave system and found that the
carbon deposits on the walls of the quartz tube could be burned off by operating
the plasma with air. However, this led to degradation and the need for replace-
ment of the quartz tube. Braun et al.[ 101 removed the carbon by addition of 0.5 -
5% O2to the helium carrier gas. This procedure allowed the determination of C
and N through atomic emission lines in the vacuum UV region. In 1972 these
authors tested a DC-based plasma detection system in combination with a gas
chromatograph. At high sensitivities the metal electrodes started to evaporate,
which made the quartz tube less bright and impeded long term stability. Moreo-
ver, the electrodes reacted with halogens.
With a microwave plasma McLean et al. [ 111 reduced the oxygen concentra-
tion to 0.1 - 1% and used spectral lines in the visible region for the selective de-
tection of halogens, H, D, C and N. They also found that nitrogen could act as a
carbon scavenger and this discovery enabled oxygen to be included in the range
of detectable elements. A commercially available system based on this publica-
tion was produced by Applied Research Laboratories in England. This system
used a low-pressure plasma, generated within an Evenson 214L resonance cav-
ity. The minimum detectable levels (MDL) were about 100 pg/s. The selectivity
of the elements relative to carbon was only about 100.
Another possibility to improve the MDLs of the low-pressure plasma, was by
increasing the plasma pressure. With the Evenson 214L cavity, used to create the
plasma, the power reflected to the microwave generator increased with the pres-
sure in the quartz tube. With this cavity it was not possible to operate at an at-
mospheric pressure. Beenakker [12,13] described a microwave cavity that is able
to work with He at atmospheric pressure. He reported lower limits of detection
between 1 and 100 pg/s. Using this type of cavity, Quimby et al. [14] in 1978,
measured limits around 10 pg/s. At that time it was not yet clear which design
was best to create a plasma. The same group [15] reported the use of a DC
plasma, using Ar as a carrier for the determination of metals. Moisan [16], and
later Abdallah [17], described a Surfatron to create a stable plasma. Our own
experiments with all four types gave rise to the following findings: the Surfatron
appeared to be very difficult to operate and high MDLs were obtained. The DC
or AC plasma did not produce sufficiently low limits of detection for non-metals.
We furthermore encountered problems with the electrodes (reaction and glow-
ing) at high power. Both the low-pressure plasma with the Evenson cavity and
the atmospheric plasma of Beenakker produced lower limits of detection, be-
tween 0.1 and 10 pg/s. For the Beenakker cavity this was also reported by Estes
et al. [18,19]. However the selectivity relative to C impeded the use of these
systems.
In order to improve this ratio (about loo), Applied Research Laboratories in
the first commercially available instrument, made an improvement by subtract-
ing a fraction of the carbon signal from the measured line (e.g. chlorine). This
improved the selectivity ratio relative to carbon to about 1000. However, the cor-
rection had to be adjusted for every emission line and moreover, the correction
was not always proportional to the carbon concentration and could only be ap-
plied over a small concentration range. Wavelength modulation with a refractor
References p . 200
164 Chapter 7
plate in the spectrometer allowed correction via nearby wavelengths on either

side of the spectral line. When a system requires short response times, e.g. for
capillary columns, the high frequencies necessary cause a decrease in signal-to-
noise ratio. The resulting loss in limit of detection compared to non-modulating
systems was demonstrated by Koirtyohann et al. [20]. The sectored wheel sys-
tem as described by McCaffrey [21] had a better signal-to-noise ratio, because
this system wasted less time travelling from the element line to the background.
However, the observation time on the element emission line is maximally 50%
of the overall measuring time, while noise is measured continuously. De Wit et
al. 1221 developed a triple-slit-exit system. This construction permitted continu-
ous and simultaneous measurement of the spectral line intensity and the back-
ground and could be used up to very short system time constants without loss
in lower limit of detection. In the next part of this chapter we will discuss a
number of parameters influencing the performance of both the low-pressure and
the atmospheric pressure plasma together with a triple-slit exit system. In 1990
Quimby [23,24] described an atomic emission detector (AED) which combined
the good lower limits of detection of the atmospheric helium plasma with
the good selectivity obtainable with diode array detection. The performance of
this system, together with some applications, will be described in sections 6
and 7.
7.4 DESCRIPTION AND EVALUATION OF A HOME-BUILT ATOMIC

EMISSION DETECTOR
This section contains a description and evaluation of the two types of micro-
wave plasma detectors used, i.e. the low-pressure plasma (LPP) and the atmos-
pheric-pressure plasma (APP) detector.
A few parameters that influence the minimum and upper detectable level
(MDL and UDL) and the selectivity will be briefly discussed. A schematic dia-
gram of the set-up for both plasmas is given in Fig. 7.1. Further details are also
given in Table 7.1.
7.4.1 Description of the apparatus
7.4.I . 1 Microwave cavities

The low-pressure plasma is created in a quartz tube placed (Fig. 7.2) in a 114
wave 214L Evenson type microwave cavity. It is provided with two tuning de-
vices in order to adjust the resonance frequency of the cavity to the frequency of
the microwave power supply, which is fixed. The quartz tube together with the
COMPUTER
2 0 ml /min SC~VE
LOW PRESSURE VACUUM PUMP

PLASMA
EXPONENTIAL DILUTION FLASK
Fig. 7.1. Schematic set-up of detection system.
cavity is mounted in a frame in order to adjust the plasma in the focus of the
spectrometer. The quartz tube is connected to a vacuum pump via a back-
pressure regulator and pressure monitor. The emitted radiation is monitored
sideways via the quartz wall of the tube.
The atmospheric-pressure plasma is generated in a quartz tube in a cylindrical
cavity (Fig. 7.3), as suggested by Beenakker, in which case the internal diameter
determines the resonance frequency. The resonance frequency is independent of
the height of the cylinder but the electric field strength increases inversely with
the height of the cylinder. Cavities with a height of 5 mm showed the best per-
formance. For a frequency of 2450 MHz with the cavity filled with air, the inter-
nal diameter is calculated to be 93.7 mm. The quartz tube is placed axially in the
centre of the cavity where the electric field strength is maximal. The introduction
of a dielectric into the cavity shifts the resonance frequency to lower values.
Therefore two tuning devices were installed and the diameter was made some-
what smaller. Cavities with 92 and 93 mm could both be tuned to a very low
level of reflected power. In later versions the coupling loop fixing block (7) was
made movable and both tuning devices removed. After one final adjustment of
the coupling loop further tuning appeared to be unnecessary and the reflected
power always remained below 0.5 %. For high-temperature work a ceramic plate
was constructed between the cavity (2) and the inlet (3). This part can be heated
References p . 200
I66 Chapter 7
TABLE 7 .I
DETAILS OF APPARATUS USED
Atmospheric plasma Cavity Beenakker type, modified depth, 5 mm

Helium flow rate 30 mVmin
Quartz tube i.d. 1.0 mm
Microwave power 50 W
Low-pressure plasma Cavity Evenson 214 L 1/4 wavelength
Helium flow rate 30 mVmin
Quartz tube i d . 1.0 mm
Microwave power 120 w
Plasma pressure 40 mbar
Lenses Material Fused silica
Diameter 38 mm
Focal length 50 mm
Monochromator Jobin Yvon HR 1000
Slit heights 15 nun
Slit widths 50 p m
Grating Holographically ruled 120 X 140mm
Lines 1200/mm
Dispersion 0.8 nm/mm
Wavelength range 180-800 nm
Photomultipliers Hamamatsu R446S or R955
Amplifiers Measuring resistance 1 MS2
Time constant 0.1 s
Scavenger Oxygen 0.2 and 0.02% v/v low and atm. plasma
Except for Hg/OP
Emission lines As given in tables
up to 450C. Air cooling is supplied via the open end (9). The cavity was
mounted on a movable support in order to adjust the plasma, which was moni-
tored via the open end of the tube, in the focus of the spectrometer.
7.4.1.2Microwave power supply
Both microwave generators are of the same type for both cavity types and in-
clude a high-voltage power supply (1600-2000 V) and a 200 W magnetron oscil-
lator, Mullard type 7090 with a resonance frequency of 2450 MHz. The power is
adjustable between 20 and 200 W. The microwave output of the magnetron is
coupled to the cavity via a reflection unit, a directional coupler Narda 3003/30
and a high power coaxial cable. When the resonance frequency of the cavity is
not exactly the same as the frequency of the generator, part of the power is re-
flected backwards into the generator and heats the magnetron. Therefore a re-
flected-current monitoring and overload protecting circuit is incorporated in or-
der to prevent damage to the magnetron if a serious mismatch occurs.
n*
COMPONENTS
SPECTROMETER
ENTRANCE
LENS
Fig. 7.2. Microwave cavity for low pressure plasma.
7.4.1.3Spectrometer
The light emitted by both the low-pressure and atmospheric-pressure plasmas
is monitored using a Jobin Yvon HR 1000 (100 cm) monochromator, equipped
with a holographically ruled grating with 1200 lines per mm. The dispersion is
0.8 d m m and the wavelength range from 180 up to 800 nm while the aperture
of the system is 1/63. The spectrometer is provided with two entrance slits and
adjustable in width from 0 to 3 mm. The desired slit can be selected by a motor-
driven mirror. It is also provided with two exit slits. One exit is the standard exit
slit of 15 mm height and adjustable in width from 0 to 3 mm. The other contains
the triple-slit exit system shown in Fig. 7.4, it consists of two small mirrors
mounted in a V-shape with an adjustable opening between the mirrors of 10-100
pm (exit slit width). These mirrors are placed in the middle of the adjustable exit
slits, thus providing two extra slits of 25-250 pm on either side of the central slit.
References p . 200
I68 Chapter 7
Fig. 7.3. Microwave cavity for atmospheric pressure cavity.
The spectral line plus background passes the central slit and is measured by the
first photo-multiplier tube. The background emission passes the side slits and is
reflected towards the second phototube. The spectral line and background photo-
tube currents are amplified and subtracted from each other.
7.3.1.3Optical system
The light emitted by the plasma is collected by two fused silica lenses with a
diameter of 38 mm and a focal length of 50 mm. The position of plasma and lens
can be changed such that a varying image from I to 3 up to 3 to 1 is projected on
the entrance slit of the spectrometer. For the atmospheric plasma a 1: 1 projection
P.M. TUBE
0
I /
/.
t-i
1 mm
Fig. 7.4. Multiple slit exit system.
is commonly used. For the low-pressure plasma a 3: 1 projection is used in order

to collect a U2.3 cone of emitted light using the F=1/6.8 spectrometer.
7.4. I.5 Sample introduction system
The gas chromatograph (GC) is equipped with both a capillary column and a
packed column and uses helium as the carrier gas. Additional helium is added to
the column exit gas to a total gas flow rate of 60 mumin for experiments where
both plasmas are used simultaneously. The low-pressure plasma is connected to
the column exit via a heated fused silica capillary (length about 30 cm, I.D. 0.22
mm) in order to suck 30 ml/min into the plasma. The remaining part goes to the
atmospheric plasma. For sensitivity and linearity measurements, the gas chroma-
tographic column was replaced by an exponential dilution flask. Both plasmas
are provided with scavenger gas supply lines, to enable the addition of various
scavenger gases (02, NZ,H2) in concentrations up to 1% of the helium. In most
experiments oxygen is used as a scavenger gas with a concentration of 2000 ppm
for the low-pressure plasma and 200 ppm for the atmospheric version.
7.4.2 Plasma start-up
The total helium flow rate through the discharge tube (column effluent, scav-
enger and make-up helium) is adjusted to the desired level, generally between 10
References p. 200
170 Chapter 7
and 50 ml/min and the microwave power set to about 50 W for the atmospheric-
pressure plasma and 120 W for the low-pressure plasma. The APP is then initi-
ated by momentarily inserting the tip of a small-diameter wire, held in a piece of
insulator, into the open end of the discharge tube. Plasma is initiated and stable
within a second. For the LPP a spark produced by a Tesla coil is used. After the
first ignition of the plasma the reflected power is turned to a minimum by mov-
ing the coupling loop a little in or out the cavity to find a position with minimum
reflected power as measured by the generator. Under normal operating condi-
tions the reflected power is less than 0.2 W at a forward power of 50 W. After a
few seconds the plasma itself has stabilized but it takes about 15 minutes for the
cavity to obtain a constant temperature and a slight retuning is sometimes re-
quired. For safety reasons the microwave radiation around the cavities was
monitored with an electromagnetic leakage monitor model 8201 of the Narda
Microwave Corporation. The stray radiation appeared to be less than 0.05
mW/cm2 at a distance of 10 cm from the cavity, which is well within the safety
limits. Only in front of the open end is the radiation level higher and the area
between lens and cavity must be covered.
7.4.3 Operating limits of the detector
Gas chromatographic detectors can be classified into two categories. For the
first group of detectors, the signal at any moment depends on the concentration
of the sample in the carrier gas. The sensitivity of these detectors, including the
well-known thermal conductivity detector, should be expressed as signal per unit
of concentration.
In the second group of detectors, the signal is dependent on the mass flow rate
of the sample. The response of the flame ionization detector, for example, is
therefore expressed in signal per mass flow rate of sample. For both plasma
types, the response is almost constant over a column flow-rate from 5 to
50 ml/min of He. This clearly indicates that the atomic emission detector is a
concentration-type detector. The minimal flow rate for the APP is limited by
upstream diffusion of air from the open end of the plasma tube.
The linear dynamic range (LDR) of the detector is the span over which the
detector exhibits a constant sensitivity to various and increasing sample concen-
trations. The lower end of this range is called minimum detectable level (MDL).
It is generally accepted that the smallest detectable amount of element is that
quantity which produces a signal equal to twice the bandwidth of the background
variations. Some parameters influencing the MDL and the LDR are discussed.
The upper detection limit (UDL) is attained when the signal is no longer line-
arly proportional to the amount of sample introduced into the detector. This up-
per limit is not a fixed point; the signal obtained begins to deviate from the ex-
trapolated linear relationship. When this deviation is more than a certain per-
centage (generally 5%), the upper limit of detection is reached. The ratio be-
tween upper limit and lower detection limit is generally expressed in decades
and should be as large as possible, e.g. larger than 4 decades. Two important pa-
rameters influencing the UDL will be mentioned, viz. the concentration of atoms
not measured in the plasma, and the type and amount of scavenger gas.
Concentrations above the UDL can be introduced into the plasma, but the sig-
nal is no longer proportional to the concentration, carbon is deposited onto the
wall of the quartz tube and finally the plasma extinguishes. Moreover, these high
concentrations also decrease the signal produced by other elements, which them-
selves are within their LDR.
7.4.3.1Emission line intensity
Helium is used as carrier gas because it has the highest excitation energy of
all elements and therefore all other elements can, in principle, be detected. Each
element that can be detected by atomic emission, has more than one emission
line between 180 and 800 nm from which of course the most intense lines pro-
duce the best MDLs (Table 7.2). Only when an emission line is too close to a
helium line is it better to use a less intense line. The intensity ratios of the vari-
ous lines of an element are constant, a criterion that can be used to increase the
reliability of the results. Except in the case of H and D, the emission lines of the
elements are well separated and interference due to overlapping emission lines
presents no problem.
TABLE 7.2
MDL AS A FUNCTION OF SENSITIVITY
Element Emission line Sensitivity h4DL

(nm) OIVlppm) (PP@
Carbon 495.8 (2nd order) 150 0.05
247.9 1880 0.006
193.1 9500 0.001
Hydrogen 656.3 2600 0.008
486.1 800 0.027
Sulphur 545.4 260 0.07
180.7 1100 0.01
Krypton 476.6 240 0.08
435.6 800 0.03
Conditions Plasma tube 1 mm i.d.
Pressure 40 mbar
Power 100 w
Scavenger, O2 3000 ppm v 0
References p . 200
172 Chapter 7
7.4.3.2Plasma pressure
With the low-pressure plasma, the intensity of the emission lines increases
with pressure, except for He and Ne whose line intensities decrease with pres-
sure. At pressures above about 200 mbar, the reflected current to the microwave
generator increases. This means that part of the power supplied to the cavity is
not absorbed by the plasma but is reflected into the generator. This reflected
power heats the magnetron and operation under these conditions is not permitted
for prolonged periods of time. Operation at plasma pressures between 80 and
120 mbar seems to be the best compromise. These conditions give excellent ac-
curacy and sensitivity. However, pressure maintenance is important.
7.4.3.3 Microwave power
With helium as a carrier gas, the plasma remains stable down to a minimum of
about 20 W. Below this power level, the plasma is extinguished with all quartz
tube diameters used (0.6 up to 6 mm i.d.). Up to the maximum power of the mi-
crowave generator (200 W) can be used. The intensity of all element lines in-
creases and the MDL decreases with power. With a 1 mm i.d. quartz tube, the
atmospheric plasma can be sustained up to about 70 W power input without
cooling. Above this power, the quartz tube melts and Si02 evaporates, which is
evident fiom a change in colour of the plasma. Without scavenger, the plasma is
OXYGEN SIGNAL, mV
QUARTZ TUBE 10 mm I D
HI FLOW R A T E 25 m l / m l n
m
I 1 I 1 I I
22 48 76 I04 I65 200
W
Fig. 7.5. S O z evaporation as a function of power.

yellow and when the SiOz evaporates, the colour changes to purple due to the
high silicon concentration. When the quartz tube is cooled, in our detector with
air or nitrogen, higher powers may be used as can be seen from Fig. 7.5. Cooling
of the quartz tube, together with not too high a power (50 W) prevents erosion of
the tube, which prolongs the lifetime to more than a month of continuous opera-
tion. Reducing erosion of the quartz tube also minimizes adsorption of compo-
nents onto the surface of the tube and improves the gas chromatographic peak
shape.
7.4.3.4Quartz tube diameter
Owing to the electromagnetic field strength in the cavity, the plasma remains
as a thin rod in the centre of the quartz tube. If the tube is too wide, i.e. more
than 2 mm, a large amount of the carrier gas passes through the tube without
passing through the plasma and is therefore not excited, resulting in a low sensi-
tivity. The APP is viewed via the open end of the quartz tube and burns at a dis-
tance of at least 5 mm from the end. Decreasing the diameter of the tube also
decreases the angle of the emitted light. For a 1 mm i d . tube, the aperture is not
much better than 1/5. With a narrower tube, a smaller cone of light is collected
and the sensitivity decreases. For both types of plasma, a 1 mm i.d. tube is the
best compromise between lifetime and sensitivity.
7.4.3.5 Optical system
In these experiments, we determined the influence of the aperture of the light
collecting system on the LDL. By varying the lenses between plasma and en-
trance slit, the cone of light collected from the plasma could be changed from
F = U2.3 up to 1/31. For the LPP, Table 7.3 demonstrates that the amount of
light reaching the photomultiplier tube (sensitivity) increases as a squared func-
tion with the aperture and the MDL improves about proportionally. The selec-
TABLE 7.3
MDL AS A FUNCTION OF THE APERTURE
Aperture Sensitivity MDL Selectivity Continuum + stray

(1/F) OIV/PPm) (PPm) ratio to C light in ppm of
element
2.7 6080 0.008 930 0.12

4.6 1832 0.010 976 0.10
6.8 905 0.013 940 0.10
10.9 3 84 0.022 946 0.10
30.8 48 0.12 940 0.10
Conditions Chlorine 479.45 nm
Slit width 50 p m
Referencesp. 200
174 Chapter 7
tivity ratio with respect to carbon remains the same for the various apertures be-
cause of the constant slit width (resolution). The last column shows the back-
ground signal due to stray light plus continuum emission from the plasma. With
an interference filter between plasma and entrance slit, we established that at
least 80% is continuum radiation from the plasma and not stray light from the
very intense helium lines. The best lower limit of detection is obtained for F/2.7
and is 0.008 ppm of CI in He. For the APP, the cone of light collected from the
plasma is not very important, whereas for the LPP, the largest possible aperture
gives the best results.
7.4.3.6Slit width
With the slit width, not only the amount of light entering the monochromator
is varied, but also the part of the spectrum (bandwidth) which is monitored.
Therefore, the effects of the slit width on sensitivity, continuum, MDL and se-
lectivity ratio to carbon were studied. For these experiments, both the entrance
and exit slit width were the same. The best lower limit of detection is obtained
using the largest aperture and 0.1 mm slit width (Fig. 7.6).
7.4.3.7 Upper limit of detection
When the concentration of an element in the plasma increases the signal of
that element increases proportionally at first. Above a certain concentration, the
increase in signal is no longer proportional to the concentration and upon further
increase of the concentration the signal levels off and the plasma can eventually
be extinguished. It is also possible to add a compound of which the signal is not
monitored, in which case the plasma is easily overloaded without visible indica-
tion. Thus, overloading should be avoided as the signal does not represent the
correct concentration and it is even possible that no signal at all is obtained. We
first tried to use the excitation temperature as an indicator for plasma overload-
ing. The excitation temperature (T',,) was calculated from the ratio of intensities
of different emission lines of the same element, in this case helium lines. At
higher temperatures, the intensity of the lines at short wavelengths were higher.
For the atmospheric and the low pressure helium plasma, we found T,,, about
3000 K, depending on conditions as power and pressure. Tanabe [25] calculated
a Text between 3 150 and 3400 K for the Beenakker cavity. The decrease in tem-
perature observed when C 0 2 was added to the plasma shown in Table 7.4.
However, the calculation of the excitation temperature takes too much time
and moreover we could not find any large differences when other parameters
were varied. Thus, it seemed much more practical to use the intensity of a he-
lium line itself as an indicator of overloading. Figure 7.7 shows the helium
587 nm line intensity as a function of the total foreign atom concentration using
the LPP. From the plot, we can see that the decrease in helium intensity occurs at
MDL
PPm
Conditions:
Low pressure plasma
tube I.D.: l.Q mm
Helium : 30 ml/rnih
Pbwer : 80 'iid
Chlorine : 479.4 nm
I
lo1 lo2
Fig. 7.6. MDL as a function of slit width.
the same total atom concentration for all atom types; and we may conclude that
the plasma starts changing when total foreign atom concentration is above 1%.
This result is in good agreement with data published by Brassem [26], who
found that as soon as the amount of the added component reaches I%, the exci-
References p. 200
I76 Chapter 7
TABLE 7.4
EXCITATION TEMPERATURE OF THE PLASMA
Type of plasma YOCOz added re,,. (K)

Atmospheric pressure 0.000 2970
0.007 2850
0.010 2780
0.014 2650
Low-pressure 0.00 2950
2.0 2680
tation conditions start to change and the line intensities are no longer propor-
tional to the concentration. The maximum concentration for the LPP is about 5%
and for the APP version about 0.5%.
7.4.3.8 Type and amount of scavenger gas
Undoubtedly, the major factor which has led to quantitative selective detec-
tion by means of a microwave induced plasma has been the use of a scavenger
gas. The plasma is created in a quartz tube, which must be kept at a temperature
well below its melting point of about 1700C. Carbon-containing compounds
60
50
+ c02
> 40 +
E
- O* A cc14
++
.-
30 +A
rn A 0 N2
3 20 O & + CH4
10
0 I , , , ,,,..I I I I I *..,I I I I . .LU
10' 1o* 10" 104 10"
Concentration ppm
Fig. 7.7. He signal for total atom concentration.

entering the plasma are pyrolysed and since carbon is hardly volatile below
35OO0C,it is to be expected that elemental carbon will deposit on the relatively
cold wall. We carefully examined the behaviour of the plasma when compounds
are introduced with and without different scavenger gases. Without any scaven-
ger, gases like 02,N2, H2, C02 and the noble gases can be added and determined
up to the maximum concentration as discussed previously. When increasing
amounts of methane are introduced, in very low concentrations ( 4 0 ppm), both
the C and H line intensities increase. At higher concentrations, the C line inten-
sity levels off, whereas the H intensity still increases. At very high concentra-
tions, the carbon is deposited onto the surface of the quartz tube and remains
there for a long period of time, continuously producing C line emission and a
continuum over the complete spectrum.
When a small amount of O2 is added to the helium, as the scavenger, methane
can be supplied up to higher concentrations without the C peak levelling out.
The overload point is about proportional to the O2 concentration. At ever higher
methane concentrations, again tailing of the C peak occurs and at still higher
concentrations, carbon is finally deposited. This carbon is burnt off by the oxy-
gen and as soon as all carbon has disappeared, the line emission of C returns to
zero. In order to maximize the linear range of the plasma detector for C the O2
scavenger concentration must be as high as possible. It cannot be increased in-
definitely, as discussed previously. N2 and H2 have the same effect on the work-
ing range of the detector. However, when carbon has been deposited onto the
wall of the quartz tube, it is not removed as quickly as with 02.The O2 concen-
trations we found to be optimal were 0.3 and 0.07% for the LPP and APP, re-
spectively. For N2 the concentrations were about 0.4 and 0.14% and H2 concen-
trations about 0.1 and 0.05%. The type of scavenger gas also depends upon the
compounds to be determined and influences the selectivity and tailing of chro-
matographic peaks.
7.4.3.9Linear dynamic range of the detector
In the previous section, we discussed the means to obtain the best MDL and
the maximum concentration of atoms that may enter the plasma. In between
these two limiting values, the LDR of the detector is found. The linear ranges
were determined for a number of elements, on a few emission lines. For these
measurements, two techniques were used in combination: viz. injecting samples
of different sizes into the GC column, (mainly used for compounds with a boil-
ing point above 50C) and the exponential dilution flask technique. The two sets
of results were then compared. The dilution vessel was installed in place of the
capillary column. For these measurements, we used 1 .O mm quartz tubes and O2
as a scavenger gas. As the detector is a concentration-type detector, the sensitiv-
ity is expressed as a signal (microvolts) per unit of concentration (ppm of the
References p. 200
1'78 Chapter 7
I 1 I I I I I
lo-* lo-' loo 10' lo2 lo3 lo4
ppm CARBON
Fig. '7.8. Linear range for carbon.
determined element in pure helium). In the graphs, this value, which is constant
in the linear range, is plotted versus the concentration of the measured element in
He. With the known gas flow rate, the weight per unit of time of component en-
tering the plasma can be calculated and a comparison can be made with known
data, e.g. from the flame ionization detector, which is mass-dependent. As the
accuracy in the lowest part of the range is low, the first decade in concentration
is not plotted.
Figure 7.8 shows the LDR for carbon using the APP with four different com-
pounds, COz, Cfi, C5H,, and CH,CI. The detection range for the latter three
compounds is linear up to a concentration of about 300 ppm of carbon in the
carrier gas. The MDL for C, using the emission line at 247.8 nm, is 0.003 ppm
so that the linear range for these compounds is about lo5.The total concentration
of foreign atoms in the plasma, at the end of the linear range, is about 2000 ppm.
The maximum for CO,, without scavenger gas, is about 600 ppm of carbon and
the total foreign atom concentration 1800 ppm. Without deposition of carbon,
the plasma can be used up to a carbon concentration of about 2000ppm, al-
though the signal is non-linear above 300 ppm. More impressive results can be
obtained at the 193.1 nm emission line, because the maximum concentration is
the same, whereas the MDL is five times better, and therefore the linear range is
also five times better.
Figure 7.9 demonstrates the difference in LDR, for argon, between the atmos-
pheric- and low-pressure plasmas. The plasmas were operated without scavenger
gas and the quartz tube diameter used was 0.6 mm i.d. The upper limit of the
linear range using the APP is about 2000 ppm, the upper limit with the LPP is
pVlvPm
Fig. 7.9. Linear range for argon.
20 000 ppm, being roughly a decade more. As we did not use scavenger gas, this
maximum concentration is also the total foreign atom concentration producing
the same values as mentioned before. The lower limits of detection for the LPP
and APP at the argon 750.3 nm line are 0.04 and 0.1 ppm and the LDRs 6 X l o 5
and 2 X 104, respectively.
7.4.3.10 Linear dynamic range for hydrogen
Most element lines exhibit a good linear relationship over an extended range.
Only a few atoms behave differently, mainly F, P and H. Fluorine appears to re-
act with the walls of the plasma-containing tube. Phosphorus is linear over only
a limited range. The flame photometric detector is a far better alternative with a
good lower limit of detection and wide range, The most important non-linearly
behaving element is H, as this atom is present in almost every component. Yet
the AED is a useful detector for H as no other selective H detector is available.
Figure 7.10 presents the sensitivity for hydrogen over a concentration range from
1 up to 5000 ppm. Up to about 1000 ppm, the sensitivity increases exponentially,
followed by a decrease due to overloading of the plasma. The same exponential
response is obtained with the LPP except that the maximum is reached at about
5000-6000 ppm. The increase in sensitivity over this range is about a factor of
two. These curves are reproducible over very long periods and are therefore still
useful in practice as long as one accepts the necessity to correct for non-linear
behaviour.
References p . 200
180 Chapter 7
30
!i 2o - CHC13
z
a
-
-___
iso-Oc term
; --- cH4
3 10
0 "
1oa 10' 10' 10' 1o4
ppm Hydrogen
Fig. 7.10. Linear range for hydrogen, LPP.
7.4.4 Selectivity
Selective detectors are very useful provided that the response of the system is
mainly due to the presence of the measured element. However, large amounts of
other compounds nearly always give a response with selective detectors. As a
measure of this interference or matrix effects, the concept of selectivity is used.
The selectivity for an element is defined as the ratio of the response per mole of
the measured element to the response per mole of another element. In GC, the
most common element is carbon, and the selectivity is often determined with
respect to carbon. As H (from hydrocarbons), 0 and N are also regularly present
in the GC effluent, the selectivity ratio to these elements was also determined.
For a limited number of combinations, we also measured e.g. the C l B r and SKI
selectivity ratios. As long as the concentration of the measured element is within
the linear range of the detector, no interference of one with the other occurs.
Only the selectivity of H versus D is rather low, as the emission lines differ very
little, being 656.28 and 656.10 nm, respectively. The selectivity ratio depends
largely on the dispersion and slit width used in the spectrometer. In the present
instrument with a dispersion of 0.8 n d m m and a slit width of 0.2 mm, the H/D
selectivity was about 10; with a slit width of 0.05 mm it was better than 500.
7 4 . 4 .I Selectivity to H, 0 and N
The selectivity ratio is not always constant over the whole concentration range
of the added element. The selectivity ratio decreases when the plasma is over-
loaded, therefore we added 1000 ppm of H, 0 and N because this is about the
maximum concentration without overloading the plasma. Selectivity ratios are
determined using the atmospheric plasma without any correction by the triple-
slit system. From Table 7.5 we can see that the selectivity for some elements
relative to H and 0 is very good (>>lOOO). The selectivity with respect to N is
not so good.
With N2 as a scavenger gas, it is necessary to use a limited concentration,
because even when the supply is constant, small variations will produce a vary-
ing background and this increases the lower limit of detection of the ele-
ments. As the ratio of H, 0, N to for example S in a molecule is never in excess
of 100, the determination of the element in a gas chromatographic peak is not
impeded.
7.4.4.2Selectivig to carbon
The selectivity to C is very important, because almost all compounds contain
C and it is most likely that an element co-elutes with much larger amounts of
carbon. Moreover, carbon produces not only line emission but also a continuum
over the complete spectrum. This continuum produces a signal at every position
of a measured elemental line. Table 7.3 demonstrates that the aperture of the
system does not have any influence on the selectivity ratio to C. The selectivity
ratio does, however, depend on the bandwidth of the spectrum that is monitored.
TABLE 7.5
SELECTIVITYTO H, 0 AND N ( X 1000)
Measured Emission Element added to plasma

element line
(nm) LPP APP
H 0 N H 0 N
C 495.8 6 15 1.5 12 14 2.0

H 486.1 - 24 0.4 - 1 0.1
c1 479.4 100 100 1.6 21 30 1.4
Br 470.5 100 100 2.2 35 80 1.3
I 516.1 100 100 7.9 100 70 10.0
S 545.4 6 3 1.o 1 7 0.6
Pressure (mbar) 20 1000
Tube i.d. (mm) 1.o 1.o
Added element (ppm) 1000 100
Power (W) 100 0
Helium (ml/min) 20 20
Referencesp . 200
Chapter 7
lo000
- F=1/31
lo00
-__- F=l/lI
--- F= 1/46
100 ....- ..... F= 112.7
10
10' 10'
Slit width JJITI
Fig. 7.1 1. Selectivity for chlorine as a function of slit width.
The selectivity improves with decreasing slit widths, as shown in Fig. 7.1 1 for
the C1479.4 nm emission line. There is not a perfect linear relationship between
slit width and selectivity because of the limited resolution of the monochroma-
tor. As the slit width also has consequences for the MDL, a compromise must be
made between MDL and selectivity. Without means to improve the selectivity,
our choice was generally a 50pm slit, because there is only a small loss in LDL
and twice the selectivity compared to a l00pm slit. In general, the selectivity
ratios for the LPP are somewhat better than for the APP.
With the triple-slit exit system, the selectivity ratios are increased by a factor
of 30-200. The precise increase in selectivity depends upon the accuracy with
which the compensation for the background variation is adjusted. This adjust-
ment can be made by injecting a pure hydrocarbon, while monitoring both the
element line emission and the background. Within the linear range of the detec-
tor, there is a constant ratio between the C signal of the background and the C
signal in the element line. In a GC analysis, both line emissions are monitored
and after the analysis, the complete chromatogram of the element line is cor-
rected for the background. When the correction factor is known, it is also possi-
ble to correct the element line for the background continuously.
When the plasma is overloaded, positive and negative peaks can occur after
correction; moreover the element signal may no longer be proportional to the
element concentration.
TABLE 7.6
PERFORMANCE OF PLASMA DETECTORS
Element Line Low-pressure plasma Atmospheric-pressure plasma

(nm)
MDL MDL Sel. Sel. LDR MDL h4DL Sel. Sel. LDR
(ppm) (PP/s) uric- xl000 xl000 (ppm) (pgk) un- x 1000 x 1000
con. corr. corr. con.
C 241.8 0.006 1.o - - 300 0.003 0.5 - - 60

193.1 0.001 0.2 - - 1000 0.001 0.2 - - 200
H 656.2 0.008 0.1 800 80 (200) 0.03 0.4 270 I0 (20)
486.1 0.04 0.6 250 20 (40) 0.17 2.4 70 20 (4)
D 656.4 0.008 0.2 600 50 (200) 0.04 1.1 100 10
0 771.2 0.13 29 40 0.038 8.4 10
N 146.9 0.13 25 1300 10 40 0.3 58 750 10 2
CI 479.5 0.009 4.4 940 100 100 0.012 5.9 950 100 8
Br 470.5 0.006 6.7 1600 40 200 0.005 5.6 620 20 20
I 516.1 0.008 14 1040 40 200 0.004 7.1 210 20 20
F 685.6 0.07 19 740 0.08 22 400
S 545.4 0.05 22 180 20 20 0.06 21 120 10 1
180.7 0.01 4.5 850 80 100 0.01 4.5 500 30 5
Hg 365.0 0.0002 0.6 105 >lo00 1000 0.0001 0.2 105 >loo0 1000
Ar 750.3 0.03 17 2900 100 0.2 100 100 7
Kr 473.8 0.03 40 460 20 50 0.15 160 140 10 4
Xe 529.2 0.008 13 4200 100 80 0.025 29 1200 100 3
Ne 640.2 0.05 14 540 50 100 0.06 17 460 30 87
e
8
184 Chapter 7
7.5 CONCLUSIONS
Table 7.1 presents the conditions used for the evaluation of both types of
plasma. A summary of the results is given in Table 7.6. For comparison, for the
MDL, the results are also given in pg/s. From these results, we may draw the
following conclusions:
1. The MDLs for both types of plasma are the same. The MDLs are between
0.1 and 100 pgs, depending on the element to be measured and on the in-
tensity of the emission line used. The MDLs of the atmospheric plasma are
almost the same as the data presented by Quimby et al. [23].
2. In order to obtain the same MDLs, the light-collecting system of the low-
pressure plasma must be better and is more expensive.
3 . The maximum concentration that can be introduced into the low-pressure
plasma is about ten times that of the atmospheric plasma.
4. The linear dynamic range is between lo3 and lo5 for the atmospheric- plasma
and between lo4 and 5 X lo5 for the low-pressure plasma.
5. The selectivity ratio with respect to carbon is about the same for both plas-
mas and ranges from 100 up to 2000 without a correction system in use. With
corrections (triple-slit system) the selectivity ratios relative to carbon are
between lo4 and lo6.
6. Because of the good sensitivity and very short. response times, both types of
plasma detectors are well suited for use with capillary gas chromatographic
columns.
7.6 DESCRIPTION OF THE HEWLETT PACKARD 5921A AED
Apart from the plasma emission detectors described in the previous

paragraphs, supplied by Applied Research Laboratories, and then by Applied
Chromatographic Systems, a number of other plasma emission detectors
have been announced by other instrument manufacturers. At the 1989 capil-
lary chromatography symposium in Riva del Garda, Italy, the Varian Instru-
ment Group (Walnut Creek, CA, USA) announced an atomic emission detector
[27] and in May 1990 Carlo Erba Instruments (Rodano, Italy) announced their
plasma emission detector ESD-4 [28] but to the time of writing, these detectors
have not been released. The only plasma detector that is commercially available
at present is the Hewlett Packard (Avondale, PA, USA) HP 5921 atomic emis-
sion detector (AED) and this instrument is described in more detail in the next
sections.
7.6.1 Gas chromatograph and transfer line
The gas chromatograph most convenient for coupling to the AED is the HP
5890A series I1 adapted with a transfer line between GC and the AED cavity. It
has a mode of monitoring and controlling the flow through the column by pres-
sure programming. The complete set-up of this GC and AED is depicted in Fig.
7.12. Since the AED is a concentration-type detector, this constant flow mode of
operation is mandatory for quantitative work. The transfer line consists of 1.5 m
of heated and insulated outer aluminium tube, with an inner tube of stainless
steel with an i.d. just over 1 mm, so that it heats the analytical column inside
properly. Since heating up this transfer line is a rather slow process, it should be
kept continuously at maximum column temperature. Up to the normal operating
temperature of 300"C, no specific measures have to be taken. In high-
temperature GC, however, where temperatures far over 300C are quite com-
mon, the limit of the temperature specification (which is 400C) is reached for
the transfer line. Neither polyimide-coated fused silica columns, nor aluminium-
clad columns can withstand these temperatures for more than 2 days (which is
50 h continuously). The only columns capable of being bent (to be pushed
1 2 3 4 5
I----
\ ' I I _- _
- A-
L
7
I n
I
I
LJ
I- I 1
I
I
6
I I
I I
/r
I
I \
I
I
I
I
I
!TI;
Fig. 7.12. Diagram of the Hewlett Packard GC-AED: (1) autosampler; (2) GC; (3) capillary col-
umn; (4) transfer line; ( 5 ) cavity; (6) magnetron; (7) spectrometer; (8) water cooling pump; (9)
cavity gas control; (10) computer.
References D. 200
186 Chapter 7
through the transfer line) which can withstand these temperatures for a long pe-
riod of time are stainless-steel high-temperature columns. Experiments have
shown that these columns only have to be replaced after 1 month of operation.
This means that the last 1.5 m of the column has been at 400C for 700 h con-
tinuously! These columns are available from Chrompack, Middelburg, The
Netherlands. The gas union of the cavity, which is the exit of the column to the
detector, as is shown in Fig. 7.13, has an i.d. of 0.75 mm, since it was designed
for acceptance of polyimide capillary columns. The steel capillary columns,
however, have an 0.d. of 0.8 mm, so the gas union should be drilled out to about
0.9 mm to accept these columns.
Fig. 7.13. The HP atomic emission detector: (1) plasma; (2) capillary column; (3) make-up and
reagent gasses in-out, solvent vent; (4) discharge tube; (5) cooling water; ( 6 ) microwave energy in;
(7) optical window; (8) window purge; (9) O-ring; (10) heated zone; (1 1) spectrometer.
7.6.2 Microwave cavity, discharge tube and gas flow system
The cavity resembles the Beenakker type, except that it has a pedestal in the
centre and a smaller diameter [23]. Microwave power is supplied to the cavity
through a waveguide of 95 mm by 45 mm, on which a microwave oven magne-
tron tube is mounted. The cavity/waveguide/magnetron assembly is attached to
the spectrometer in such a way that it allows the cavity to be swung away for
discharge tube replacement and repositioning. The cavity contains a quartz dis-
charge tube of 1.0 mm i.d. X 1.25 mm 0.d. X 42 mm long, which is water-
cooled to eliminate erosion. The lifetime of the discharge tube as measured, is at
least 1 month. Since columns were replaced every month, and replacement of the
column can only be performed by disassembling the heated part of the cavity, the
discharge tubes were replaced at the same time. The exit of the cavity is closed
with a UV-grade fused silica window and purged with helium to prevent back
diffusion of ambient air. It also enables reversion of gas flows inside the dis-
charge tube to allow solvent venting.
The cavity contains a 70 W cartridge heater, enabling heating of the cavity
block up to 400C. Coupling the analysis column to the cavity block is per-
formed through the gas flow system. This system allows the entrance of one or
more auxiliary gases (He, H2, 02,N2, CH,), or venting of sample solvent, as
controlled by the computer. The use of these gases prevents peak tailing greatly,
depending on the nature of the peak. A recipe for the proper use of these gases is
incorporated in the computer software.
When a sample is to be injected by the on-column injection technique, it gen-
erally has to be diluted before injection, The amount of diluted sample injected
into the column will then be in the order of a few microliters, and this large
amount of solvent will extinguish the plasma, or even worse, cover the inner sur-
face of the discharge tube with a layer of carbon deposit. This layer of soot can
only be removed by the plasma itself, which takes many hours. For these analy-
ses, a solvent vent is a very convenient part of the design. The choice made in
the design of this detector to cool the discharge tube with water, instead of a gas-
cooling system, has some important practical consequences (see Fig. 7.14 for the
detailed construction):
1. To prevent a significant consumption (and thus loss) of microwave power
by the cooling water, the water film is only 100,um thick. This calls for
filtering of any particulates out of the water and prevention of the forma-
tion of algae, so a water-filtering system is essential.
2. To cool the discharge tube effectively, the linear velocity of the very thin
water film must be rather high. Together with the filtering system, it de-
mands a powerful pump and a flow sensor to stop the microwave power if
the flow drops below a predetermined value.
References p . 200
I88 Chapter 7
It 12
Fig. 7.14. Construction of the HP AED cavity with water cooling: (1) pedestal; (2) discharge tube;
(3) quartz jacket; (4) water in; ( 5 ) out; (6) polyimide ferrule; (7) cover plate; (8) makeup and rea-
gent gas inlet and vent outlet; (9) heat stand 0% (10) heated zone; (1 l ) column; (12) column fer-
rule; (1 3) gas union; ( 1 4) collar; (1 5) water plate; (16) coupling loop.
3. In those cases where high column temperatures are necessary, there is a

rather large temperature difference between the gas union and the water
plate. This temperature drop is effected over the polyimide ferrule, which
has a triple function: (a) it isolates the water system from the gas system;
(b) it isolates the cavity overpressure from the outside; and (c) it fixes the
discharge tube to be kept in place. The demands put on the ferrule are ex-
cessive when temperatures over 300C are needed.
The coefficients of expansion of the stainless steel of the gas union and the
polyimide of the ferrule are quite different. When using temperatures over 300C
and switching off the block heater, which is e.g. performed by the flow sensor in
the case of too high a restriction in the water lines, the ferrule will crimp more
than the gas union. Cooling water will commence to leak along the ferrule and
sometimes into the electronics compartment. Complete disassembly of the cav-
ity, replacement of the ferrule and repositioning of the discharge tube are then
necessary. The polyimide of the ferrules in use for more than one week at 400C
appear very much deteriorated at inspection. After installation of a fresh ferrule,
the cavity needs partial re-assembly after a day, to retighten the ferrule. A redes-
ign of this part of the detector for high-temperature work should therefore be
considered.
7.6.3 The photodiode array spectrometer
The light emitted by (the elements in) the plasma passes a helium-purged
quartz window. It passes the 0.05 mm entrance slit of the spectrometer and is
diffracted and focussed by a concave holographic grating with a flat focal plane
of 0.35 m and sent to a 21 1 pixel photodiode array (PDA) [24]. This PDA moves
along the focal plane, which is nearly linear in shape from 160 to 800 nm. The
optic arrangement is depicted in Fig. 7.15. Simultaneous measurements of lines
more than 10-30 nm apart (depending upon their position in the focal plane) is
impossible. Multiple chromatographicruns are necessary in those cases.
The spectrometer is capable of measuring clustered lines within the width of
the PDA simultaneously. But it can also be set at any wavelength and it can scan
a continuous spectrum. It includes both polychromator and monochromator fea-
Fig. 7.15. Optics schematic diagram of the HP AED:(1) helium plasma; (2) 90' elliptical mirror;
(3) entrance slit; (4) concave diffraction grating; ( 5 ) PDA, (6) focal plane.
References p . 200
190 Chapter 7
tures. The spectrometer itself is heavily thermally insulated and since wave-
length correction is performed at the start of each chromatographic run, it means
that the thermal time constant of wavelength precision is in excess of 20 h. The
cavity is attached to the outside wall of the spectrometer, so that the focus of the
elliptical mirror is 2 mm into the end of the discharge tube. The 21 1 detecting
elements of the PDA are all active 100% of the time, so that signal and back-
ground portions next to a spectral line are continuously measured and corrected
for.
7.6.4 Computerized control and data treatment
The gas chromatograph as well as the AED is computer controlled with a HP

35920A GC-AED Chemstation. All functions of the system can be performed by
softkey control. Correct scavenger gases are automatically turned on, and the
required order sorter (optical filter) is chosen. It automatically calibrates the
AED for wavelength determinations before each run.
To locate the atomic line centres with the PDA precisely, the centre of gravity
calculation algorithm [29] is used. For most atomic lines, it results in a standard
deviation of the estimation of the line centre better than 0.01 pixel. The helium
spectrum provides several narrow, intense lines most suited as wavelength stan-
dards. The auxiliary gases provide small amounts of carbon, nitrogen, hydrogen
or oxygen, which also can be used as calibration lines. The mechanical position
of the PDA on the focal plane is converted to nominal wavelength (which is very
close to linear), using a numerical model of the spectrometer. Since selectivity of
non-metallic elements in atomic emission spectroscopy relative to carbon rarely
exceeds a few hundred, background correction is necessary to improve this. An
improvement by several orders of magnitude is obtained by means of real-time
multipoint background correction. See also Table 7.7.
The PDA read-out time is usually 10 Hz,but sometimes 100 Hz is used during
calibration routines. Signals specific for an element are produced by a computer-
induced recipe, composed of a pair of linear functions of the various pixel sig-
nals. In this recipe, pixel responses, element filter and background signal are re-
corded, so that the calculated background amount to give optimal selectivity can
be ad-justedpost-run.
7.6.5 Characteristics
The sensitivities and the selectivities of the various elements depend on the
element line used for measurement. A number of common elements on which we
Microwave pfasma detectors 191
TABLE 7.7
SENSITIVITIESAND SELECTIVITIESOF SOME COMMON ELEMENTS WITH THE
HEWLE'IT PACKARD AED
~ ~~
Element Wavelength DL ( P m a Selectivity (relative to carbon)
Nitrogen 174 10 6000

Sulphur 181 <2 150000
Carbon 193 <1
Carbon 248 2
Hydrogen 486 2
Carbon 496b 10
aTo enable comparison of the AED with e.g. the FID, the MDL is expressed in pg/s, where the
AED is a concentration-type detector and the MDL should basically be expressed in a concentra-
tion unit.
bAlthough carbon 496 is a 2nd order line of carbon 248, it is generally used when both hydrogen
and carbon have to be determined.
determined these characteristics, all exhibited better selectivities and sensitivities

than the specifications, as supplied with the instrument, suggested. Some typical
examples are given in Table 7.7.
7.7 TYPICAL APPLICATIONS
7.7.1 Multi-element SimDist analyses
The use of gas chromatography for simulated distillation (SimDist) analysis

and its implications are described elsewhere in this volume. For the specific
analysis of multi-element SimDist, especially on residue-containing products,
some practical considerations are discussed.
To enable the elution of very high-boiling material from the column, short
(typically 5-10 m) wide-bore capillary columns are used. These columns contain
a very thin film (typical 1100 nm) of a chemically bonded, non polar stationary
phase, stable at high temperatures (typical 44OOC).
As mentioned in the previous chapter, the AED is a concentration-type detec-
tor and quantitative results can only be obtained when the flow is strictly con-
stant. The flow of mobile phase (helium) through the column is generally about
10 ml/min, which means that the column pressure drop is around 15 P a . Since
the cavity pressure, caused by auxiliary gases and window purge gas, is about
15 H a , the column head pressure is about twice the pressure drop of the column.
The HP 5890A series II gas chromatograph is equipped with a so-called con-
stant-flow module. However, this constant flow is obtained by pressure pro-
References p . 200
192 Chapter 7
r-'--7- I --- T-----7-----7----. . . 7-

0 10 20 30 40
Tfme <min.>
Fig. 7.16. C, H and S chromatograms for SimDist analyses of a long residue.
gramming. When this constant-flow mode is used without any further considera-
tions. the electronics will programme the pressure assuming that the column
head pressure equals the column pressure drop. This will result in a non-linear
flow programme during analysis. The solution here is to calculate the necessary
pressure programme to provide constant flow and to introduce the results of
these calculations via the keyboard into the GC electronics.
Generally, samples on which a SimDist analysis is performed have a wide
boiling range and on-column injection is the only mode of injection to be used
here. As mentioned previously, solvent vent should be used to prevent the huge
amount of solvent entering the AED. Depending on the column starting tempera-
ture, co-eluting, low-boiling parts of the sample will be vented as well. With a
typical wide-bore column of 6 m length and a starting temperature of 3OoC, the
initial boiling point of the sample must not be below 150C. Programming the
column up to 430C will enable compounds with normal boiling points up to
700C to be eluted. Repeated experiments demonstrated that the temperature of
the transfer line and cavity should not be more than 30C below the maximum
oven temperature, otherwise high-boiling compounds will not be detected com-
pletely.
7.7.1.1 Multi-element SimDist software
Hewlett Packard provides a software package specifically written to perform
SimDist analyses with the AED. It consists of a set of macros that execute the
II I
Fig. 7.17. Boiling point calibration with the AED using normal alkanes.
w
w
various program files used. These include boiling point calibrations, calibrations
for the correction of the carbon response on other element lines (back-amounts),
response calibrations as well as blank run determinations. The package enables
the user to set up their own element set, to choose which elements should be de-
termined. Reports can be produced that give the mass of an element recovered,
as well as the hydrogen-to-carbon ratio, as a function of the boiling range, pre-
sented in a table as well as graphically, in either degrees Celsius or degrees
Fahrenheit. It automatically determines and controls all analytical runs that have
to be performed. These include all calibration runs as well as runs necessary to
detect the specified elements.
Calibrations to determine the back-amounts and responses of specific ele-
ments have to be performed by injection of synthetic mixtures including one or
more compounds containing these elements. In a future release of the software,
the possibility of calibration with real oil samples will be incorporated. As in
general most hetero compounds are distributed throughout the boiling range, this
kind of calibration can be expected to be more accurate than single-compound
calibration.
Some publications suggest that the AED exhibits a compound or structure de-
pendency of response [30]. We have determined the response of the AED to-
wards carbon and hydrogen for a set of different structured organic compounds.
The results are given in Table 7.8. Clearly, dependency on structure is not de-
References p . 200
194 Chapter 7
'TABLE 7.8
STRUCTURE INDEPENDENCY ON RESPONSE FOR THE HP AED
Compound Formula Percentage of hydrogen (C + H = 100%)
Theor. DCPDSa r1-Cl2~ Naphthalenea
2-Methyl-hexadecane 15.0 15.1 15.2

2-Methyl- 1 -dodecene 14.3 14.2 14.2
1-Methyl-decalin 13.2 13.3 13.3
I , 1-Dimethyl-tetralin 10.1 10.2 10.1
Tetramethy I-benzene 10.4 10.4 10.4
Fluoranthene 4.9 4.9 4.9
1,4-Dithiane 14.4 14.4 14.5
Quinoline 6.1 6.0 5.9
Dimethyl-sulfolane 14.4 14.4 14.4
~~~~~ ~~ ~
"DCPDS, calibrated with dicyclopentyl dodecyl sulphide; n-C 12, calibrated with n-dodecane;
naphthalene, calibrated with naphthalene.
monstrable. These results have all been obtained using a strictly constant column
flow (by means of a proper pressure programme; see also Section 7.7.1) and
taking the non-linearity of the hydrogen response into account. More about this
hydrogen non-linearity is described in the next paragraph.
1.7.1.2 Linear dynamic ranges
Quimby and Sullivan [23] present an extensive description of the HP AED
design, including a table with selectivities, sensitivities and linear dynamic
ranges (LDR). In order to be able to perform Multi-Element SimDist of the ele-
ments H, C, S and N, we determined the ability of the AED to determine these
elements. Although the minimal detectable level for nitrogen given by Quimby
and Sullivan is 7.0 pg/s, this low level is hard to reach in high-temperature GC
use. In normal practice of high-temperature GC, it is almost impossible to ex-
clude minor leaks in injection port (septa) and column couplings, so that a con-
stant relatively high background of nitrogen is always present. Since nitrogen-
containing compounds in oil samples generally are distributed along the boiling
range, nitrogen is present in thousands of compounds. The maximum concentra-
tion of nitrogen rarely exceeds a few thousand parts per million. Thus, the car-
bon-to-nitrogen ratio of eluting material is in general over 10 000, and equals or
even exceeds the selectivity of 6 X lo3 given in 1231. For these reasons, the AED
is not the detector of choice for SimDist of the element nitrogen in high-boiling
oil samples.
Hydrogen exhibits a deviant behaviour in terms of linearity, as is depicted in
Fig. 7.18. We have observed this phenomenon on different plasma detectors (see
1.5
1.4 u
1.3
1.2
1.1
$5 l.o
;
$ 0.7
0.6
0.5
0.4
h
1 2 3 5
1 5 10 5 10 5 10 5 10 5 10
Fig. 7.18. Non-linearity of hydrogen.
also Section 7.4.3), but have not yet been able to produce a conclusive explana-
tion. The shape of the linearity curve is indeed highly reproducible and can
therefore be determined exactly. The Multi-Element SimDist software includes a
method to measure this non-linearity and calculate response factors accordingly.
It should be realized, however, that these response factors vary with the abun-
dance of the hydrogen line intensity, and therefore vary along the hydrogen peak.
In normal peak integration mode, as was the case with the measurements of Fig.
7.18, these factors cannot be incorporated and thus hydrogen will not be meas-
ured correctly. Implementation of these factors is automatically performed by the
SimDist software, since it does not integrate separate peaks, but rather integrates
and treats the chromatogram in narrow area slices. It generates a LDR for H486
of about 10 000. Together with the LDR of C496 and S181 of over 10 000, it pro-
vides an excellent detector for SimDist analysis for these elements.
7.7.1.3 Simdist results
In normal SimDist analyses with the flame ionization detector, where only
carbon is measured, no response factors are used. For samples containing a high
concentration of polynuclear aromatics, too high a recovery will therefore be
reported, whereas very highly paraffinic samples will exhibit too low a recovery.
Apart from these errors, hetero elements (such as sulphur) present in the sample
References p . 200
196 Chapter 7
TABLE 7.9
COMPARISON OF ELEMENT RATIOS WITH THE HP AED
YOC %H %S
Comb. AED Comb. AED Comb. AED
Sample A 12.1 12.8/12.9 85.5 84.8B4.8 2.40 2.36/2.35

Sample B 12.0 13.0112.8 85.0 83.9184.2 3.03 3.1 U2.98
Sample C 11.5 12.6/12.4 87.7 86.3B6.5 1.03 1.06/1.09
SampleD 13.8 14.3113.8 86.2 85.6/86. I <0. 1 0.06/0.04
SampleE 13.9 13.9/1 3.6 86.1 86.1/86.4 <0.01 0.015/0.015
will not be detected at all. When using the AED, none of these errors will occur.
These two types of SimDist results should therefore be compared with this con-
sideration in mind. Since the carbodhydrogen ratio determined with the AED is
a good yardstick for the estimation of the amount of paraffinic and (po1y)-
aromatic material in a sample, we were able to select a number of samples for
this comparison. The conclusion is that differences in boiling points reported for
1.64-
Y8 CIS
I 1
N r n
0 0
1.60-
!i
t
In
6
m
W 1.55-
9
.
1.45-
1.4@
_, - --*
Ii
1 I I
0.00 5.00 10.00 15.00
Retention time In minutes
Fig. 7.19. Chlorinated components in the EO process.

carbon and hydrogen, will in general not be larger than 3C. The AED results of
the totals of the different element distributions and the comparison with the re-
sults obtained by combustion analyses of total samples are presented in Table
7.9.
7.7.2 Determination of chlorine-containingcompounds in the ethylene oxide

process
In the ethylene oxide (EO) process, ethylene is converted to ethylene oxide

over a catalyst. Traces (1 ppm) of chlorinated components are added to tune this
catalyst. Figure 7.19 is a typical analysis of the reactor outlet using the C1 chan-
nel of the home-built apparatus.
7.7.3 Determination of oxygenates in gasoline
Modem non-leaded gasolines contain oxygenates as octane boosters. Figure

7.20 represents the 0 and C chromatograms of such a gasoline using the Hp
atomic emission detector. See also the work of Wylie et al. [31].
7.7.4 Determination of noble gases in natural gas
In oil-producing reservoirs, tracers are sometimes introduced via the steam

injector and samples are taken from the producing wells in order to identify
preferential gas or steam paths. The noble gases are preferred because they are
chemically inert and non-radioactive. Figure 7.21 represents a neon chroma-
togram of a synthetic mixture using the home-built apparatus.
7.7.5 Determination of metal porphyrins in crude oils
For the detection of nickel, vanadium and iron porphyrins in crude oils, the
reader is referred to the work of Quimby et al. [32] and Zeng ei al. [33].
7.8 CONCLUSION
At the time of writing, the only commercially available plasma detector is the
Hewlett Packard 5921A Atomic Emission Detector. This detector has been de-
References p . 200
198 Chapter 7
5 000
4 000
3Q0Q
jl.;
Z@OQ
1 El00
. . *
2 4 6
?
R
Time ( m i n . )
o 777 o f DRTR:CMDPSTRC.D
1400-
Q 2 .1 6 8
Time (rnin.)
Fig. 7.20. Carbon and oxygen chromatogram of gasoline.
signed such that it is compatible with capillary columns up to temperatures of

400C. It is a very good compromise between performance, ease of operation
and price. The choice of a moveable diode array as spectrometric detection de-
vice enables the determination of a few elements simultaneously, if the emission
lines are within the range of the array. For elements with emission lines more
apart, the automatic GC sampler enables multiple analyses in an accurate man-
ner.
5.40-
5.36-
5.31-
5.27-
5.23-
.01
Fig. 7.21. Natural gas, 90 ppb neon added.
To decrease erosion of the discharge tube, which contains the helium plasma,
the system is water cooled. The construction of the cooling of the cavity makes it
very sensitive towards water leakages at temperatures above 300C. Apart from
this source of trouble, the detector in combination with the Chemstation software
is a powerful and easy-to-use device. It provides the user with menus in which
the gas chromatograph as well as the detector can be controlled automatically,
calibrated and used for analysis.
The detector enables measurement of both the elemental composition of
eluting compounds as well as element distributions of complex samples. Ex-
periments pointed out that the response of the detector for the various elements
is structure independent.
In our laboratory, the detector has been used successfully for the determina-
tion of a wide variety of elements: viz. metals, such as lead, tin and mercury, the
noble gases as neon, krypton, argon and xenon; oxygen-containing compounds,
where it successfully replaces the OFID; halogen-containing compounds, where
it replaces the EC detector; sulphur-containing compounds, replacing the FPD;
and last but not least, hydrogen, for which no other selective detector is avail-
able. In most of these cases, the MDL of the AED is the same or even better than
that of the other selective detectors. Furthermore, the AED in general is imme-
diately ready for use for detection of a new element, whereas installing a new
detector needs quite an investment in stabilization time and calibration.
Although the detector has only been available since 1990, a number of inter-
esting applications have been published already and it is to be expected that
many more will follow.
200 Chapter 7
7.9 REFERENCES
1 H.P. Broida and G.H. Morgan, Anal. Chem. 24 (1952) 799.

2 A.J. Mc Cormack, S.C. Tong and W.D. Cooke, Anal. Chem. 37 (1 965) 1470.
3 C.A. Bache and D.J. Lisk, Anal. Chem. 37 (1965) 1477.
7 H.A. Moye, Anal. Chem. 39 (1967) 1441.
8 R.S. Braman and A. Dynako, Anal. Chem, 40 (1968) 95-106.
9 R.M. Dagnall, S.J. Pratt, T.S. West and D.R. Deans, Talanta 16 (1969) 797.
10 W. Braun, N.C. Peterson, A.M. Bass and M.J. Kurylo, J. Chromatogr. 55 (1971) 237.
11 W.R. McLean, D.L. Stanton and G.E. Penketh, Analyst 98 (1973) 432.
12 C.I.M. Beenakker, Spectrochim. Acta 31B (1976) 483.
13 C.I.M. Beenakker, Spectrochim. Acta 32B (1977) 173.
14 B.D. Quimby, P.C. Uden, R.M. Barnes, Anal. Chem. 50 (1978) 21 12.
15 R.J. Lloyd, R.M. Barnes, P.C. Uden and W.G. Elliot, Anal. Chem. 50 (1978) 2025.
16 M. Moisan, R. Pantel, J. Microwave Power 14 (1975) 57.
17 M.H. Abdallah and S. Coulombe, Spectrochim. Acta. 37B (1982) 583.
18 S.A. Estes, P.C. Uden and R.M. Barnes, Anal. Chem. 53 (1981) 1829.
19 Estes, P.C. Uden and R.M. Barnes, J. Chromatogr. 239 (1982) 181.
20 S.R. Koirtyohann, E.D. Glass, D.A. Yates, E.J. Hindenberger and F.E. Lichte, Anal. Chem. 49
(1977) 1121.
21 J.T. McCaffrey and R.G. Michel, Anal. Chem. 55 (1983) 488.
22 A. de Wit, Spectrochim. Acta 38B (1983) 369.
23 B.D. Quimby and J.J. Sullivan, Anal. Chem. 62 (1990) 1027.
24 J.J. Sullivan and B.D. Quimby, Anal. Chem. 62 (1990) 1034.
25 K. Tanabe, Spectrochim. Acta 36B (1981) 633.
26 P. Brassem, F.J.M.J. Maessen and L. de Galan, Spectrochim. Acta31B (1976) 537.
27 Proceedings of the Tenth International Symposium on Capillary Chromatography 989, ol-
ume 11, 1025 pp.
28 Advanced Product Release, Carlo Erba Instruments, Rodano, Italy, May 1990.
29 J.A. Stewart and A. Scheeline, Anal. Chem. 56 (1984)
, , 2995.
30 S. Pedersen-Bjergaard, T.N. Asp and T. Greibrokk, Anal. Chem. 15 (1992) 89.

31 B.D. Quimby, P.C. Dryden, and J.J. Sullivan, High Resolut. Chromatogr. Chromatogr. Com-
mun. 14 (1991) 110.
32 P.L. Wylie and B.D. Quimby, High Resolut. Chromatogr. Chromatogr. Commun. 12 (1989)
813.
33 Y . Zeng, J.A. Seeley, T.M. Dowling, P.C. Uden and M.Y. Khuhawar, High Resolut. Chroma-
togr. Chromatogr. Commun. 15 (1992) 669.
E.R.Adlard (Ed.), Chromatography in the Petroleum Industry
Journal of ChromatographyLibrary Series, Vol. 56
0 1995 Elsevier Science B.V.All rights reserved 20 1
CHAPTER 8
The sulfur chemiluminescence detector
Richard S. Hutte
Sievers Instruments Inc., 2500 Central Avenue, Suite HI, Boulder, CO 80301, USA
8.1 INTRODUCTION
All petroleum samples from crude oil to refined products contain varying
concentrations of sulfur-containing compounds. These compounds include gases
(H2S, COS, SO2 and CS2), aliphatic, and aromatic thiols, sulfides and polysul-
fides, thiophenic and other sulfur-heterocyclic compounds. The geochemistry of
sulfur has been recently reviewed [ 13 and an extensive list of the types and con-
centrations of sulfur compounds found in petroleum systems is presented. The
concentration of sulfur compounds in crude oils varies widely from <0.05% to
>14% sulfur by weight [l]. Most of the sulfur is in the form of organic com-
pounds, with only relatively small amount of dissolved H2S and elemental sulfur
present. Most of the organo-sulfur compounds are higher molecular weight com-
pounds with boiling points greater than 300C [l]. Hydrotreating and other proc-
esses to remove sulfur compounds from crude oil and other feedstocks result in
the conversion of the higher molecular weight materials to H2S and lower mo-
lecular weight thiols and sulfides. Thus, a wide range of different types of sulfur
compounds is usually present in petroleum products.
Sulfur compounds are also intentionally added to petroleum products. Low
molecular weight thiols, sulfides and tetrahydrothiophene are used as odorants
for natural gas and LPG and certain additives for lubricants and other products
contain sulfur.
The presence of even parts per billion levels of sulfur can cause numerous
problems in the processing of petroleum including poisoning of the expensive
catalysts used in the refining process, corrosion of reactors and pipelines and the
presence of sulfur-containing compounds imparts undesirable odors to petroleum
products. Combustion of petroleum products containing sulfur compounds is a

202 Chapter 8
major source of SOz air pollution and acid rain. For these reasons, accurate de-
termination of the concentrations and identities of the sulfur-containing species
in petroleum products is required.
Many analytical techniques have been developed for the measurement of the
sulfur content of petroleum and petroleum products. These techniques include
methods for the determination of total sulfur content including X-ray fluores-
cence [2], coulometric [3] and radiometric [4] techniques and chromatographic
techniques that permit the quantification of individual sulfur-containing com-
pounds. The most widely used chromatographic technique is gas chromatogra-
phy, usually coupled with sulfur-selective detectors.
8.2 SULFUR-SELECTIVEDETECTORS FOR GAS

CHROMATOGRAPHY
The most widely used sulfur-selective detector for gas chromatography has
been the flame photometric detector (FPD) IS]. Sulfur-selective detection in the
FPD is based on combustion of sulfur-containing compounds in a hydrogen-
ricldair flame to produce diatomic sulfur in an electronically excited state (&*).
The emission from Sz* is monitored using a photomultiplier tube positioned near
the flame. The problems of the FPD for sulfur detection are well documented [6]
and include a non-linear response, compound-dependent response and perhaps
most important for petroleum analyses, quenching of the response due to co-
elution of hydrocarbons. Despite these limitations, the FPD has been success-
fully used for the determination of sulfur compounds in a wide range of petro-
leum samples for many years [7-1 l].
The Hall electrolytic conductivity detector (HECD) can also be used in a sul-
fur-selective mode [ 12,133. Sulfur compounds are combusted in the presence of
oxygen in a heated nickel tube to form SOz,which is bubbled into a suitable sol-
vent such as methanol or methanol/water and the electrical conductivity of the
solvent is monitored. The HECD is not widely used for sulfur-selective detec-
tion, in part because it is viewed as being difficult to operate and maintain and
suffers from interferences and quenching of the sulfur response.
The atomic emission detector (AED) is a multi-element detector that can be
used for sulfur compounds [14-16]. The AED has a linear response for sulfur
compounds and does not suffer from quenching, but has poor selectivity for sul-
fur versus carbon [ 161 that limits detection of low levels of sulfur compounds in
hydrocarbon matrices. The AED is also relatively expensive and requires highly
skilled operators.
The analysis of low levels of sulfur by gas chromatography is complicated by
the reactivity of most sulfur compounds. Sulfur compounds are irreversibly ad-
The chemiluminescence detector 203
sorbed or will decompose on many surfaces, especially heated metal surfaces.

This results in the loss of sulfur compounds in sample collection equipment and
throughout the chromatographic system (syringes, injection ports, columns, and
detectors). Thus, precautions must be taken to minimize sorption and loss of low
level sulfur compounds in the samples by replacing metal surfaces with glass,
fused silica, Teflon, or less reactive metals such as nickel, and by conditioning
the chromatographic system by injection of high concentrations of the sulfur
compounds of interest. Sorption and loss are particularly noted for H2S, SO2 and
mercaptans, while sulfides and disulfides are less reactive.
Preparation of standards containing sulfur compounds is also a problem. For
gas standards, low level sulfur compounds can be lost on the walls of the gas
cylinders, requiring the use of specially treated cylinder and stabilization of gas
mixtures. For liquid standards, chemical reactions such as oxidation of mercap-
tans by air to form disulfides occurs even with refrigeration of the solutions.
8.3 THE SULFUR CHEMILUMINESCENCEDETECTOR
The limitations of existing sulfur-selective detectors for gas chromatography

led to the development by Benner and Stedman [17] of a new sulfur-selective
detector, the sulfur chemiluminescence detector (SCD).
Combustion of sulfur compounds in a hydrogen-rich flame produces a number
of different sulfur species including HIS, HS, S, S2, SO and SO2, with the ratios
of the various species determined by the fuel/air ratio and other factors [6]. Ben-
ner and Stedman [I71 noted that sulfur monoxide (SO) is one of the major spe-
cies formed in the combustion of sulfur compounds in reducing flames and, in
fact, is present at 10 times higher concentration than atomic sulfur. Detection of
SO produced in a flame should provide a more sensitive and linear detection
system for sulfur compounds.
Halstead and Thrush [18] reported in 1966, that sulfur monoxide undergoes a
chemiluminescent reaction with ozone. Emission in the blue and ultraviolet re-
gion of the spectrum (260480nm) was observed from the emission of elec-
tronically excited SO2 formed from the reaction of SO with 0,.
The SCD combines these reactions to provide sensitive and selective detec-
tion of sulfur compounds. Sulfur monoxide is produced by combustion of sulfur
compounds in a hydrogen-rich flame, the flame gases are collected by means of a
sampling probe and transferred to a chemiluminescent reaction chamber where
they are mixed with ozone and the emission monitored using a photomultiplier
tube after passing through a wide band-pass filter.
H,lair flame
Sulfur compounds > SO + products

204 Chapter 8
so + 0 3 -3 so2+ o2+ hv
The SCD developed by Benner and Stedman is an atmospheric monitor for
the measurement of both oxidized and reduced sulfur compounds in ambient air.
The detector has a linear response for sulfur compounds, equal molar response
and a detection limit of 0.13 ppbv (-1 pg S/s). No quenching of the sulfur re-
sponse is observed due to C 0 2 and hydrocarbons and this lack of interference
provides for improved measurement of low parts per billion level sulfur com-
pounds in ambient air compared with fluorescence and FPD sulfur monitors
1191.
At Sievers Instruments, we investigated the use of the SCD as a chroma-
tographic detector by coupling the Benner and Stedman SCD with a gas chroma-
tograph. These preliminary experiments led to the development of a commercial
detector for gas chromatography, the Model 3 50 Sulfur Chemiluminescence
Detector. A schematic of the SCD is shown in Fig. 8.1. It consists of a hydrogen-
rich flame, a flame sampling probe, a transfer line, an ozone generator, a chemi-
luminescent reaction cell, an optical filter, a photomultiplier tube and associated
electronics for detection of the chemiluminescent radiation and a vacuum pump.
In the GC-SCD, the flame source is a conventional flame ionization detector
operated under hydrogen rich conditions. The flow rates of hydrogen and air for
the flame in the SCD are 200 ml/min and 400 ml/min, respectively. In contrast,
normal FID flow rates are typically 30 mumin for hydrogen and 250-400 ml/min
for air. The more reducing flame results in the formation of sulfur monoxide
from the combustion of sulfur compounds, yet still provides a flame ionization
detector response, although the sensitivity of the FID is reduced by approxi-
mately an order of magnitude due to the higher hydrogen flow rates.
The flame sampling probe in the SCD is a high purity ceramic tube 8-1 1 cm
in length (0.5 mm i.d.) positioned -4 mm above the FID jet. To avoid collection
of room air, only -90% of the flame gases are collected by the ceramic probe.
The probe is positioned in the flame using a bayonet mount to lock it into a mov-
able mounting base shown in Fig. 8.2. The position of the probe in the flame can
be optimized by either adjusting the position of the mounting base or rotating the
probe handle to one of three locking positions, thus repositioning the end of the
probe relative to the flame jet.
The vacuum pump of the SCD serves several purposes; collection of the flame
gases, transfer of the flame gases and ozone into the chemiluminescent reaction
chamber, and reduction of non-radiative collisional quenching of the emitting
species (SO2*) in the chemiluminescent reaction cell. Under normal operating
conditions, the chemiluminescent cell pressure is 10-1 5 Torr. A chemical trap is
used to remove ozone and oxides of nitrogen from the gas stream before the vac-
u u m pump and since the flame gases contain high concentrations of water vapor,
MODEL 350 B SCD
REGULATOR
FILTER
RESTRICTOR
OZONE
GENERATOR
VACUUM
TRANSFER UNE MNT
SAMPUNQ
188EYBLY
GAS or
<FIO - -1
I
I
I
I
I
SUPERCRITICAL I
I
FLUID I
ELECTRONICS
CHROMATOGRAPH I
B 3 RECORDER
Fig. 8.1. Schematic of the sulfur chemiluminescencedetector.

RECORDER
the pump is operated with the gas ballast open to vent water from the pump. To
prevent loss of oil, an oil coalescing filter is used to collect oil in the pump ex-
haust and the oil is returned to the pump.
The transfer line consists of a 1.5-m length of 4.75 mm (3/16 inch) 0.d. black
perfluoroalkoxy (PFA) tubing. The use of an inert material minimizes loss of
sulfur monoxide in the transfer line and the relatively large inside diameter re-
References pp. 22 7-229

206 Chapter 8
To Chemiluminescence Detector
Ceramic Probe
Fig. 8.2. Schematic of the fixed position interface for mounting the SCD probe on an FID.
duces condensation of water. Residence time in the transfer line and chemilumi-
nescent reaction cell is approximately 1 s and no significant band-broadening or
peak tailing is observed.
Emission from the SO/03 reaction occurs in the blue and ultraviolet region of
the spectrum, therefore a blue-sensitive photomultiplier tube is used to detect the
chemiluminescent reaction. The flame gases also contain relatively high concen-
trations of nitric oxide, which will also undergo a chemiluminescent reaction
with ozone. Emission from the N 0 / 0 3 reaction occurs in the red and near-
infrared region of the spectrum and therefore an optical filter that transmits ra-
diation over the wavelength region of 240410 nm is positioned in front of the
PMT to eliminate interference from nitric oxide.
In the commercial SCDs, two different techniques have been used to process
the signal from the photomultiplier tube. The original detector used photon-
counting, while newer versions employ a pico-ammeter. The principal advantage

of the analog system is a wider dynamic range and minimal distortion of the
chromatographic peak shape from the signal processing electronics.
8.4 PERFORMANCE CHARACTERISTICSOF THE SCD
The SCD has a linear response for sulfur compounds over five orders of
magnitude, a specified detection limit of <5 pg S/s [20] with a detection limit of
<0.5 pg S/s having been reported [21]. This sensitivity corresponds to -10-50 pg
of a sulfur compound or parts per billion concentrations, depending upon the
sample size, injection technique (split versus direct or splitless), and other chro-
matographic variables. The selectivity for sulfur compounds versus hydrocar-
bons of >lo6 [20-221. The SCD response is equimolar for sulfur compounds and
the response is not quenched by co-elution of higher levels of non-sulfur-
containing compounds.
The excellent selectivity of the SCD is achieved primarily by the combustion
of the sample in the flame. Compounds that will undergo chemiluminescent re-
action with ozone, such as olefins, are converted to C02 and water, eliminating
possible interference. However, under certain circumstances, a positive hydro-
carbon response is observed. For example, when a large amount of a compound
elutes as a very sharp chromatographic peak, the compound is not completely
burned in the flame and the partial combustion products can react with ozone
and produce a response. In this situation, it is necessary to either decrease the
sample size or to change the chromatographic conditions to broaden the chroma-
tographic peak to permit complete combustion of the components.
An SCD response is also observed for arsine, phosphine and their organo-
derivatives, although the sensitivity of the SCD for these compounds has not
been determined. Nitriles also produce a small SCD response, presumably due to
CN emission however this response is much lower that the response for sulfur
compounds.
The equimolar response of the SCD for sulfur compounds has been demon-
strated for a wide range of sulfur compounds [21-241. The relative response
factors for a range of sulfur compounds (relative to phenyl sulfide) are shown in
Table 8.1. As will be described in more detail in the applications of the SCD, the
equimolar response permits the determination of the total sulfur content (or total
volatile sulfur content) by simply summing the individual components, or when
only total sulfur content is desired, then a short chromatographic column, with
poor resolution can be employed and the overall detector response integrated for
the determination of total sulfur content.

208 Chapter 8
'TABLE 8.1
RELATIVE RESPONSE FACTORS FOR SULFUR COMPOUNDS USING THE SULFUR
CHEMILUMINESCENCE DETECTOR
Compound Relative response factoP
H2S 0.80
cos 0.90
so1 1.oo
CS2 1.oo
Methanethiol I .43
Ethanethiol 1.18
I -Propanethi01 1.05
2-Propanethiol 1.04
1-Butanethiol 1.11
1 -Methyl- 1-propanethi01 1.02
2-Methyl- 1-propanethi01 1.05
2-Methyl-2-propanethiol 1.11
1-Pentanethiol 1.12
1-Hexanethiol 1.08
I -Heptanethiol I .oo
1-0ctanethiol 1.06
1-Decanethiol 1.04
Dimethyl sulfide 1.oo
Methyl ethyl sulfide 0.88
Diethyl sulfide 1.07
2-Methyl-2-propane sulfide 1.02
Dimethyl disulfide 0.95
Diethyl disulfide 1 .oo
Thiophane I .04
Thiophene 1.08
2-Methy lthiophene 1.10
3-Methyl thiophene 1.oo
2,j-Dimethyl thiophene 1.05
2-Ethyl thiophene 1.03
2-n-Propyl thiophene I .02
2-n-Butyl-thiophene 1.oo
Benzothiophene 1.05
Dibenzothiophene 0.87
"Relative to phenyl sulfide, data from [21-241.

8.5 FACTORS INFLUENCING THE SENSITIVITY AND SELECTIVITY

OF THE SCD
The key step in the SCD is the combustion of sulfur compounds in the hydro-
gen-rich flame to form sulfur monoxide and collection of SO by the sampling
probe. Therefore flame conditions and the position of the probe in the flame are
the most critical factors in determining the sensitivity, selectivity, and stability.
The hydrogen/air ratio in the flame determines the products of sulfur com-
bustion. For the best SCD sensitivity and stability, the hydrogen flow rate is set
to -200 ml/min and the air flow rate set to -400 mVmin or a fuel/air ratio of
-1 :2. Under these conditions, the tip of the sampling probe is heated to sufficient
temperatures that a dull red glow can be observed when the probe is rapidly re-
moved from the flame. These conditions also yield sulfur monoxide as the major
product of sulfur combustion.
With higher air flow rates >400 ml/min (or lower hydrogen flow rates) sulfur
dioxide is the principal combustion product and the SCD sensitivity is decreased.
The higher air flow rates also produce a hotter flame as indicated by a white
glow at the tip of the probe, when removed from the flame.
At lower air flow rates (<400 mumin), the yield of SO in the flame is actually
increased, but several other factors make operation at lower air flow rates unde-
sirable. Whenever the sampling probe is in the flame and the ozone generator is
on, a background signal is observed for the SCD. In part, this background is due
to the so-called ozone-wall reaction, a background radiation observed when
ozone is passed in front of a photomultiplier tube. However, an additional back-
ground signal is observed from the reaction of the flame gases (possibly hydro-
gen atoms) with ozone. Under the normal hydrogen and air flow rates, this back-
ground signal is relatively constant and not affected by the elution of non-sulfur-
containing compounds from the GC However, when the flame for the SCD is
operated at lower air flow rates, this background signal can be quenched by
the elution of non-sulfur-containing compounds from the GC. This results in
negative peak for these components when the SCD is operated with low air flow
rates.
In addition to the negative SCD response for non-sulfur species, operating the
flame of the SCD at lower air flow rates produces a cooler flame (no glow ob-
served for the tip of the probe) and this lower temperature does not produce sta-
ble response over a period of several days. Thus, any increase in sensitivity
achieved at the low air flow rates is offset by the reduced stability observed.
Thus, the optimum conditions for most GCs are very close to the 200 ml/min
hydrogen flow rate and 400 ml/min air flow rate.
Examples of the SCD response obtained at different air flow rates are shown
in Fig. 8.3. The sample is a test mixture containing five sulfur compounds (and

210 Chapter 8
N O %x.Q473
RESPONSE
n
h
Fig. 8.3. Effect of hydrogen and air flow rates on sensitivity and selectivity of the SCD. (A) Chro-
matogram of sulfur standard with low air flow rate. (B) Chromatogram of sulfur standard with cor-
rect hydrogenhir ratio. (C) Chromatogram of sulfur standard with high air flow rate.
some impurities) in hexane solvent, with 10% benzene. Figure 8.3B shows the
SCD chromatogram obtained under the proper gas flow rates (200 mumin H2,
400 ml/min air). No response is observed for -130pg of hexane (2pl split 1O:l).
Under the chromatographic conditions employed, benzene co-elutes with thio-
phene, thus slightly distorting the thiophene peak on the SCD, however no
quenching is observed. Figure 8.3A shown the same test mixture using
200 ml/min H2 and 350 mumin of air. Under these more reducing conditions, a
negative response is observed for hexane and benzene and a greater response of
the detector for the sulfur compounds is obtained. Finally, Fig. 8.3C shows the
SCD response for the same test mixture using 200 ml/min H2 and 500 ml/min of
air. The response for the sulfur compounds is greatly reduced and in some cases,
a positive response for the hydrocarbons can be observed under these more oxi-
dizing conditions. This example illustrates the importance of operating the detec-
tor at the proper gas flow rates.
When the SCD is operated at the proper gas flow rates, good day-to day re-
producibility can be obtained, however, contamination of the probe from column
bleed and septa bleed can cause a loss in sensitivity. Silicone compounds from
either GC columns or septa when high injection port temperatures are used can
deposit on the tip of the probe and decrease the response of the detector for sul-
fur compounds. In most cases, the contamination can be physically removed by
simply inserting a cleaning wire into the probe. To avoid the silicone bleed con-
tamination, the columns (and septa) should be well conditioned and the column
operated at as low of temperature as possible for a given analysis. For example,
most bonded methyl silicone columns can be operated at temperatures up to
275C, without any bleed problem, but operation at higher temperatures such as
300C can cause contamination of the probe and reduced sensitivity. It should be
noted that the SCD is more sensitive than the FID to silicone bleed and contami-
nation can occur even though no significant baseline rise is observed on the FID.
8.6 FLAMELESS SULFUR CHXMILUMINESCENCE
A dedicated ceramic burner for the formation of sulfur monoxide has been re-
cently described by Shearer [25] and a schematic of the burner is shown in Fig.
8.4. In this flameless SCD, hydrogen and air are mixed with the column efflu-
ent in a heated ceramic tube and all of the combustion gases are transferred to
the chemiluminescence detector. The typical gas flow rates for the burner are
100 mumin for hydrogen and 20 ml/min of air, which are outside the flammabil-
ity limits for H2/air. To initiate and sustain the combustion, the burner is heated
typically to 800-900C and direct connection of the chemiluminescence detector
(and vacuum pump) to the burner results in a reduced pressure, typically

212 Chapter 8
HYDROGEN INLET
1/16' CERAMIC
HEATING ELEMENT HYDROGEN DELIVERY TUBE
REACTION ZONE
1/8" CERAMIC TUBE
THERMOCOUPLE
COLUMN CONNECTION
Fig. 8.4. Schematic of enclosed burner for the conversion of sulfur compounds'[o sulfur monoxide.
-200 Torr in the ceramic tube. This modification of the SCD improves the ease
of use of the SCD, by eliminate the need to position a fragile ceramic probe in an
FID and also results in improved sensitivity. Detection limits of 2-5 fg S/s were
obtained by Shearer [25].The improved sensitivity is due in part to the more
reducing conditions of the combustion, increasing the yield of SO and lower
pressures in the chemiluminescent reaction cell, reducing non-radiative colli-
sional quenching of SO2*. As with the flame-based SCD, a nearly equimolar re-
sponse is observed for sulfur-containing compounds and good sulfur to carbon
selectivity (>lo8) is obtained [25].
The burner is available commercially, however the sensitivity is somewhat re-
duced compared with Shearer, with typical detection limits of >0.5 pg S/s re-
ported [26]. One drawback of the lower gas flow rates of the ceramic burner is
that quenching of the sulfur response due to co-elution of hydrocarbons can be
observed. In most cases, this quenching can be reduced or eliminated by increas-
ing the hydrogen and air flow rates, while maintaining the 5: 1 hydrogerdair ratio
(e.g. 200 ml/min H2, 40 mVmin air), however decreased sensitivity for sulfur
compounds is obtained at these higher flow rates. For example, the sulfur re-
sponse is decreased by approximately a factor of two at 200 ml/min H2,
40 ml/min of air versus the response obtained at 100 ml/min H2, 20 mVmin
of air. Another drawback of the ceramic burner is that in order to obtain simul-
taneous SCD/FID signals it is necessary to use a post-column split. However,
for many routine applications and for low level detection of sulfur com-
pounds, the ceramic burner offers many advantages compared to the flame-based
SCD.
8.7 COLUMN SELECTION AND SAMPLING TECHNIQUES
As previously noted, sulfur compounds can be sorbed and lost in all compo-
nents of the chromatographic system and in sampling containers. Specially
treated gas bombs and cylinders have been developed to minimize the loss of
sulfur compounds, but in most cases, passivation of the sampling devices by
treatment with high levels of sulfur compounds is required to avoid losses of low
levels of sulfur compounds. In many cases, the best sampling containers are
older ones that have been in use for long periods of time and thus have been
passivated through use. Gas-tight syringes can also be a source of problems.
Some syringes are more active than others and cause significant loss of low lev-
els of sulfur compounds. When high levels samples are analyzed, the sulfur
compounds can permeate through the Teflon and other syringe components, then
slowly outgas, resulting in contamination of future samples analyzed with this
syringe.
Passivation with high levels of sulfur compounds is also the most common
method for minimizing loss in the chromatographic system. Exposing inlet lines,
gas sampling valves, sample loops, and injection port liners to high levels of
H2S, SO2, mercaptans and other reactive sulfur compounds can reduce the loss of
sulfur compounds in the GC system. Decomposition and loss of sulfur com-
pounds can also occur in the chromatographic column. For example, porous

214 Chapter 8
- 6 4 mV dibenzothiophene
ETU 3 n-oc tadecanethiol
0 YlnuU.
5 10 16 20 25
5m x 0.53mm 1.5umu DB-5 FSOTC (J&W SC.)

35OC t o 430% Q 12.O0/min.
r dibenzothiophene
rthianthrene
n-octadecanethiol
A
0 Mlnutsr 5 10 16 20
5m x 0.53mm O.lOum,, SPB-1 FSOTC (Supelco)

50C to 43OoC Q 14.Oo/min.
Fig. 8.5. Illustration of decomposition of sulfur compounds in a fused silica capillary column.
polymers provide good separation of volatile sulfur compounds but low levels of
sulfur compounds can be lost in the column and no amount of passivation ap-
pears to overcome this problem. The same is true for PLOT columns and the best
materials we have found for analysis of low levels of the sulfur gases are the
treated silicas such as Chromosil.
Decomposition and loss of sulfur compounds can also be observed in fused
silica capillary columns. An example of the decomposition of ethylene thiourea
(ETU) on a megabore column with a relatively thin film is shown in Fig. 8.5.
Analysis of a standard mixture of four-sulfur containing compounds ETU,
dibenzothiophene, thianthrene and n-octadecanethiol using a column with a
1.5p m film thickness shows good peak shape for all of the sulfur compounds.
Analysis of the same mixture using a column with a 0.1 p m film thickness
shown almost complete decomposition of the ETU and severe tailing of the other
sulfur compounds. Sorption and loss of reactive sulfur compounds like ETU has
been observed even for new fused silica capillary column, indicating that the
loss of the reactive compounds is most likely due to interactions with the silica
surface and not simply due to an active column. This suggests that whenever
possible, the analysis of sulfur compounds should be performed using fused sil-
ica column with film thickness of l p m or greater.
A particularly useful capillary column has been developed for the analysis of
sulfur compounds. The column (30 m X 0.32 mm i.d. SPB-I 4,um film thick-
ness, Supelco Inc.) provides separation of H2S from co-eluting COS/S02 at 35C
and can be used for samples up through the diesel range. At -1O"C, COS and
SO2 can be separated. Methyl silicone columns with 4 ,urn film thickness are now
also available from other column manufacturers. The combination of a thick film
methyl silicone and special cross-linking to minimize column bleed permits
separation of most sulfur compounds and applications for a wide range of petro-
leum samples.
The combination of high sensitivity and selectivity of the SCD has led to the
development of a number of applications for this detector in the measurement of
sulfur compounds in petroleum and petroleum products. Some representative
examples are given below.
8.8 APPLICATIONS
8.8.1 Refinery gases
A major application for the analysis of sulfur compounds is natural gas, LPG,
refinery gases and other process gas streams. Since these streams are usually
upgraded or sold for heating purposes, accurate measurement of the levels of
sulfur compounds is required. Figure 8.6 shows the FID and SCD chroma-
tograms obtained from the analysis of a refinery gas (LP) sample using the
flame-based SCD. A 0.1-ml sample was injected onto a capillary column, using a
split injection technique (split ratio 1O:l). The FID chromatogram shows that
propylene and propane are the major constituents of the sample with low levels
of C4 and C5 hydrocarbons. Hydrogen sulfide is the major sulfur contaminant,
with lower levels of COS, SO2, mercaptans, sulfides disulfides and thiophenes
present in the sample.
Not all of the sulfur compounds were identified, however, the equimolar re-
sponse of the SCD for sulfur compounds can be used to determine the sulfur
content of these unidentified components and the total sulfur content of the
sample. A response factor (pg S/area) can be determine from the analysis of a
standard containing one or more sulfur compounds and this response factor used
to quantitate unknown sulfur compounds and total sulfur content can be calcu-
lated from the total area.
Another important application is the measurement of sulfur compounds in
polymer grade ethylene, propylene and other olefins used in the production of
plastics. Due to the complications from low levels of sulfur compounds in the
polymerization reactions, the desired level of total sulfur in the feedstock is

216 Chapter 8
Gas Chomatograph Hervlet-Packard Model 5890

Sulfur Detector Slevers lnst, Inc Model 3506 SCDbn
Cokmn 30m x 0 32mm 4p SPBl FSOT (Supelco. Inc )
Column Temperature 3 mn @ -10% to 3ooOC @ 10 0 Ohin
Carrier Gas Hekum Q 22 png (3 0 d h n )
Speaal Spkt lryector 250% Split vent Row 32 c d h n
FID 300OC kr @ 39 psig H2@ 60 psig
Makeup Air Q 30 cm?lMn
Manual Injecbon 0 1 mL gas (8 6 O h to column)
SCD Response
FID Response
0 2 4 6 0 10 I2 14 I6 18 20 22 24 26 28
00605018
Yhrtaa
Fig. 8.6. SCD and FID chromatograms of a refinery gas

Calibration mixture
80 ppb carbonyl sulfide
40 ppb hydrogen sulfide

80 ppb sulfur dioxide
20 ppb carbon disulfide

t
SCD
SCD
FID
I
1 I I I I I
0 min 2 4 6 8 10
Fig. 8.7. Analysis of polymer grade propylene using the SCD.
<5 ppb. To facilitate detection of sulfur compounds at the low ppb levels using
the SCD, larger sample sizes and direct injection onto a packed column can be
used. Figure 8.7 shows the analysis of a calibration standard and a sample of
polymer grade propylene using a Chromosil 310 column. As noted above, the
treated silica packed in a Teflon column provide a sufficiently inert system for
the detection of low ppb levels of sulfur gases. For this propylene sample, the
levels of the individual sulfur compounds were <20 ppb.

218 Chapter 8
8.8.1.2Gasoline
The concentration of total sulfur in motor gasolines typically ranges from 300
to 500 mg S k g (ppm). The development of new reformulated gasolines is now
underway in an effort to improve ambient air quality. Included in the regulations
governing fuel reformulation are new limits for the concentration of total sulfur
in the gasolines. For example, the California Air Resources Board regulations
call for a total sulfur content of less than 40 mgkg in gasoline. These lower sul-
fur concentrations present many problems, not only in the refining and blending
of the fuels, but also to the analytical chemist.
The SCD and FID chromatograms obtained from the analysis of a sample of a
commercial motor gasoline is shown in Fig. 8.8. This oxygenated gasoline is
used in the winter months in the Denver, Colorado, area as part of a program to
reduce atmospheric carbon dioxide levels. The SCD chromatogram shows the
typical sulfur compounds present in gasoline, thiophene and alkyl thiophene,
benzothiophene and the alkyl derivatives. In addition, lower levels of thiols and
sulfides are also present in this sample, with a total sulfur content of 337 mg
S/kg. The FID chromatogram shows the typical hydrocarbon pattern expected for
gasoline, aromatic and aliphatic hydrocarbon, and also shows methyl tert-butyl
ether (MTBE), the additive used for the oxygenated fuel program.
An example of the SCDKID analysis of a reformulated fuel is shown in Fig.
8.9. The SCD chromatogram again shows the thiophenic sulfur compounds, but
lower levels of thiols and sulfide, with a total sulfur content of 36 mg S/kg. The
FID chromatogram from the reformulated fuel shows MTBE and higher levels of
2,2,4-trimethylpentaneand other iso-paraffins compared with typical gasolines.
8.8.1.3Diesel fuels
The total sulfur content of diesel fuels is also subject to current and pending
limitations, with proposed limit of 500 ppm. Some example of the sulfur and
hydrocarbon profiles for diesel fuels are shown. The FID and SCD chroma-
tograms from the analysis of NIST Standard Reference Material No. 1624b
(Sulfur in Distillate (Diesel) Fuel Oil) are shown in Fig. 8.10. This material is
certified to contain 332 f 3 mgkg sulfur and is useful for calibrating the SCD
for total sulfur determinations. Benzothiophene, dibenzothiophene and alkyl de-
rivatives of these compounds are the major sulfur components of this sample and
most diesel fuels. For comparison, the FID and SCD chromatograms for a low
sulfur diesel (26.5 mg S/1) are shown in Fig. 8.1 1 .
8.8.1.4High temperature gas chromatography
High temperature GC is widely used to determine the boiling range of crude
oils and petroleum feedstocks. The flame-based SCD permits simultaneous de-
termination of the hydrocarbon and sulfur compounds boiling range distribu-
Gas Chromatograph: Hewlett-Packard Model 5890

Sulfur Detector: Sievers Inst., Inc. Model 3506 SCD'm
Column: 30m x 0.32mm 41.1 SPBl FSOT (Supelco, Inc.)
Column Temperature: 1 min. @ 35OC to 3OOOC @ 10.0 O h i n .
Carrier Gas: Helium @ 22 psig (3.0 cm3/min.)
Special Split Injector: 250C Split vent flow: 32 cm3/min.
FID: 300C Air @ 39 psig. H, @ 60 psig.
Makeup: Air @ 30 cm3/min.
Manual Injection: 2.5pL (8.6% to column)
SCD Response
u
loo 1 t
o !
0 5 10 15 20 25
minutes
FID Response
0 5 10 IS 20 25
Minutes
Fig. 8.8. SCD and FID chromatogramsof a commercial gasoline.

220 Chapter 8
Gas Chromatograph: Hewlett-Packard Model 5890

Sulfur Detector: Sievers Inst., lnc. Model 3508 SCDtm
Column: 30m x 0.32mm 4p SPBl FSOT (Supelco, Inc.)
Column Temperature: 1 rnin. @ 3 5 O C to 3OOOC @ 10.0 O/min.
Carrier Gas: Helium @ 22 psig (3.0 cm3/min.)
Special Split Injector: 250C Split vent flow: 32 cm3/min.
FID: 3OOOC Air @ 39 psig. H, @ 60 psig.
Makeup: Air @ 30 cm3/rnin.
Manual Injection: 2.5pL (8.6%to columnj
SCD Response
I
O !
3 1 10 IS 20 23
mutes
FID Response
0 1 13 13 20 23
mrts
Fig. 8.9. SCD and FID chromatograms of a reformulated gasoline.

The chemiluminescence detector 22 I
Gas Chromatograph: Varian Model 3410

Sulfur Detector: Seven Inst., lnc. Model 350A SCDm
Column: 1Om x 0.53mm 1 . 5 ~DB5 FSOT (J&W Scienbfic)
Column Temperature: 0 min. Q 40C to 4OOOC Q 10.0 Vmin.
Carrier Gas: HeliumQ 2 psig (6.0 CmVtnin.)
SPI Injector: 100C - 400CQ 20*C/min.
FID: 400C Air Q 40 psig. HZQ 35 psig.
Makeup: Helium Q 30 cm3/min.
Manual Injection: 0.2pL (Hamiiton #lOOl)
SCD Response
60 - 0
f
c
a Calibration Analysis
.-f NIST SRM #1624b
- 8
40
Sulfur in Distillate (Diesel) Fuel Oil
mV
Sulfur Content: 0.332k 0.003 mass YO
20 - Density @ 6GF: 0.8628 g/cm3
0
0 4 8 12 16 20 24 28 32
Ylnuirr 20828021
FID Response
0 4 8 12 16 20 24 28 32
Yinulrr 20828020
Fig. 8.10. SCD and FID chromatograms of NIST SRM No. 1624 sulfur in distillate (diesel) fuels
oil.
222 Chapter 8
~~~~~~~~ ~

Sulfur Detector Sieves Inst.. Inc. Model 350A SCDbn
co(LgMI: 10mx0.53mm 1.5~085 FSOT (J&WScienttfic)
Column Temperature: 0 min. Q W< to 400% Q 10.0 O/rnin
Carrier Gas: Helium Q 2 psig (6.0 & h n . )
-
SPI Injector: 100C 400C @ 2CFC/min.
FID: 400C Air Q 40 psig. Hi Q 35 psig.
Makeup: Helium Q 30 crr?hin.
Manual Injection: 0.2pL (Hamilton #7001)
SCD Response
d l
Total Sulfur = 28.5 mas&
0 4 8 li :F 20 24 28 32
Y in D i e 8 20828011
FID Response
60
40
mv
21)
0
0 4 8 12 I6 20 24 28 32
Yiauiev 2082801 R
Fig. 8.1 1. SCD and FID chromatograms of a low sulfur diesel fuel.

Sulfur Detector: Sievers Inst., Inc. Model 350A SCDm
Column: 10mx0.53mm 1.w DB5 FSOT (Jaw Scientific)
Column Temperature: 0 min. Q 400C to 4OOOC Q 10.0 Ohin.
1 Carrier Gas: Helium Q 2 psig (6.0 cm3hin.)
-
SPI Injector: 100C 400C Q 20"Chin.
FID: 400C Air Q 40 psig. H, Q 35 psig.
i Makeup: Helium Q 30 c d h i n .
I Manual Injection: 0.2pL (Hamilton #7001)
SCD Response
30 1
20
MV
10
0 4 a 12 16 20 24 28 32 36
Ylnulrr ?Oh27081
FID Response
'0 1
mV 50 -
0 4 S 12 16 20 24 28 32 35
Yinutea 20827088
Fig. 8.12. SCD and FID chromatograms of West Texas intermediate crude oil.

724 Chapter 8
1 Sulfur Detector: Seven Inst.. Inc. Model 350A SCD"

j j Column: 10mx0.53mm 1 . 5 ~DB5 FSOT (Jaw Scientific)
1I Column Temperature: 0 min. Q 40% to 400C Q 10.0 */min.
~! Carrier Gas: HeliumQ 2 psig (6.0 cm)hin.)
~!SPI Iqector: 100C - 400C Q MoChnin.
i j FID: 400C Air Q 40 psig. I-$Q 35 pslg.
, Makeup: Helium Q 30 c r n h i n .
' Manual Injection: 0.2pL (Hamilton W o o l ) i
SCD Response
75 1 al
-
u
5
511 - Total Sulfur = 4.471 mg/L
mV
25 -
FID Response
0 4 8 I2 16 20 24 28 22 36
Y In u t s s 20827098
Fig. 8.13. SCD and FID chromatograms of Ordovician crude oil.


Sulfur Detector: Sievers Inst.. Inc. Model 350A SCDm
Column: 10m x 0.53mm 1 . 5 ~DB5 FSOT (JawScientific)
Column Temperature: 0 min. Q 4OoC to 4OOOC Q 10.0 Ohin.
Carrier Gas: HeliumQ 2 psis (6.0 CrrPMn.)
-
SPI injector: i0OC 400C Q 20"Chin.
FID: 400C Air Q 40 psig. H, Q 35 psig.
Makeup: Helium Q 30 crrhtnin.
Manual Injection: 0.2pL (Hamilton #7001)
SCD Response
Feedstock to Hydrotreater Unit
'i
3 -
Total Sulfur 5 1.813 maS/k
.I
2 -
1 -
0 4 6 12 16 20 24 26 ,'
1
.
ih 40 41
Y in "1. I 20621361
Product from Hydrotreater Unit
3 .
.
I
2.
Total Sulfur = 204.8 maSlL
1 -
0 - 7
Fig 8.14. SCD chromatograms of gas oil before and after hydrotreating.

226 Chapter 8
tions. As previously noted, the SCD is susceptible to contamination and loss in

sensitivity due to column and septa bleed and this problem is particularly acute
in high temperature GC. The use of well-conditioned columns and low bleed
septa are required for high temperature GC with SCD detection. With these pre-
cautions, the SCD can be used for the simultaneous hydrocarbon and sulfur
compound boiling range distribution.
Figure 8.12 shows the FID and SCD chromatograms from the analysis of
a West Texas Intermediate crude oil. The major sulfur components are di-
benzothiophene and its alkyl derivatives. There is also a relative large amount
FID Response
Feedstock to Hydrotreater Unit
15
mY
lb
- .
0 4 8 12 16 23 24 28 32 36 40 44
Yinrtmm 20828068
Product from Hydrotreater Unit

25 1
o l
0 4 E 12 16 20 24 28 32 36 40 44
Ylnrtmm 20828088
Fig. 8.15. FID chromatograms of gas oil before and after hydrotreating.
of higher molecular weight (bp > C30)sulfur compounds present in this sample
as indicated by the unresolved hump observed at the end of the chromato-
gram.
An even higher concentration of these high molecular weight sulfur com-
pounds can be seen in Fig. 8.13, which shows the FID and SCD chromatograms
from an Ordovician Crude Oil. The total sulfur content of this sample was found
to be approximately three times than the West Texas crude and a large percent-
age of the sulfur compounds are higher molecular weight species.
The SCD can also be useful in monitoring refinery processes by measuring
the concentrations of sulfur compounds in feedstocks and resultant products. An
example of this is shown in Figs. 8.14 and 8.15 for the feedstock and product of
a hydrotreating unit. The SCD chromatograms shown in Fig. 8.14 illustrate that
significant reduction in the concentrations of sulfur compounds is achieved by
this treatment, while the FID chromatograms, Fig. 8.15, indicates that the major
hydrocarbon constituents are relative unchanged.
8.9 CONCLUSIONS
The high sensitivity, selectivity and linear response of the sulfur chemilumi-
nescence detector have made this instrument a valuable tool for the measure-
ment of sulfur compounds in petroleum and petrochemicals. The equimolar re-
sponse for different sulfur compounds and the absence of quenching of the re-
sponse due to co-elution of hydrocarbons permits determination of low ppb lev-
els of sulfur compounds in hydrocarbon matrices and determination of total sul-
fur content.
The SCD has also found applications in the measurement of sulfur com-
pounds for foods, flavor, and beverages, pesticides and other environmental
applications, detection of chemical warfare agents, quality control for pharma-
ceutics and basic chemicals and a host of other areas [27-331.
8.10 ACKNOWLEDGMENTS
The author would like to thank Neil Johansen of Sievers Instruments for pro-
viding the chromatograms and his assistance in the preparation of the chapter.
8.11 REFERENCES
1 W.L Orr and C.M. White, Geochemistry of Sulfur in Fossil Fuels, American Chemical Soci-
ety, Washington DC (1 990).
228 Chapter 8
2 ASTM Standard D2622, Annual Book of ASTM Standards, Vol. 05.02, ASTM Philadelphia,
PA (1990).
PA (1987).
PA (1 990).
5 S. S. Brody and J. E. Chaney, J. Gas Chromatogr. 4 (1966) 42.
6 S. 0. Fanvell and C. J. Barinaga, J. Chromatogr. Sci. 24 (1986) 483.
7 M. Dressler, in: Selective Gas Chromatography Detectors, Elsevier, Amsterdam (1986), p.
133.
8 C. Bradley and D. J. Schiller, Anal. Chem. 58 (1986) 3017.
9 J. L. Buteyn and J. J. Kosman, J. Chromatgr. Sci. 28 (1990) 19.
10 T.R. McManus, Anal. Chem. 63 (1991) 48R.
1 1 R.S. Hutte and J.D. Ray, in: Detectors for Capillary Chromatography, Wiley, New York
(1992), p. 193.
12 R.C. Hall, J. Chromatogr. Sci. 12 (1974) 152.
13 R.C. Hall, in: Detectors for Capillary Chromatography, 109.
14 P.C. Uden, Y. Young, T. Wang and Z. Cheng, J. Chromatogr. 486 (1989) 319.
15 P.C. Uden, in: Detectors for Capillary Chromatography, Wiley, New York (1992), p. 219
16 B.D. Quimby and J.J. Sullivan, Anal. Chem. 61 (1990) 1027.
17 R.L. Benner and D.H. Stedman, Anal. Chem. 61 (1989) 1268.
18 C.J. Halstead and B.A. Thrush, Proc. R. SOC.London 295 (1966) 363.
19 R.L. Benner, R.L. and D.H. Stedman, Environ. Sci. Technol. 24 (1990) 1592.
20 Operation and Service Manual Model 350B Sulhr Chemiluminescence Detector, Sievers
Instruments, Inc. (1990).
21 R.L. Shearer, D.L. ONeal, R. Rios and M.D. Baker, J. Chromatogr. Sci. 28 (1990) 24.
22 H.V. Drushel and G.D. Dupre, Sulfur compounds in petroleum by GC/SCD: detector
evaluationioptimization and BP/TR correlations, presented at the 1991 Pittsburgh Conf-
erence.
23 ASTM Committee D-2 Proposed Standard Test Method for the Determination of Sulfur Com-
pounds in Petroleum Gases and Light Liquids by Gas Chromatography and Chemilumines-
cence Detection.
24 K.J. Bohler, A.J. McCormack and J.M. McCann, Simultaneous detection of aromatics, sulfur
and hydrocarbons in diesel fuels by gas chromatography, presented at American Chemical
Society Meeting (1991).
25 R.L. Shearer, Anal. Chem. 18 (1992) 2192.
26 Operation and Service Manual Model 355 Sulfur Chemiluminescence Detector, Sievers In-
struments, Inc. (1 993).
27 N.G. Johansen, R.S. Hutte and M.F. Legier, in: Monitoring Water in the 1990s: Meeting New
Challenges ASTM STP 1102.
28 H.C. Chang and L.T. Taylor, Anal. Chem. 63 (1991) 490.
29 M.S. Burmeister, C.J. Drummond, E.A. Pfister and D.W. Hysert, J. Am SOCBrew. Chem. 50
( 1992) 53.
30 R.S. Hutte and R.E. Sievers, Selective detection of HD and VX using the sulfur chemilumi-
nescence detector, presented at 47th Southwest Regional American Chemical Society Meeting
( 1991).
3 1 K. MacNamara, Investigation of medium volatile sulfur compounds in whiskey, presented at
the Cognac Symposium (1992).
32 R. Dominguez, Jr., The determination of total sulfur in fuel, landfill and sewage digester gas
using the sulfur chemiluminescence detector, presented at American Chemical Society Meet-
ing (1992).
33 R.L. Shearer, E.B. Poole and J.B. Nowalk, J. Chrornatogr. Sci. 31 (1993) 82.
0 1995 Elsevier Science B.V. AU rights reserved 23 1
CHAPTER 9
Multi-column systems in gas

chromatography
H. Mahler, T. Maurer and F. Mueller

Siemens AG, AbtAUT35 CHR, Postfach 21 12 62, 76187 Karlsruhe. Germany
9.1 INTRODUCTION
9.1.1 Limitations of single-column systems
Unquestionably, most practical applications of gas chromatography can be

solved with a single-column system. However, laboratories in the petroleum in-
dustry often have to analyse samples that may contain several hundred com-
pounds. A complete separation of such mixtures with a single-column system is
impossible and even very high resolution capillary columns with efficiencies of
many thousands of theoretical plates are not sufficient to provide full resolution.
Using longer and longer columns will not solve the problem because it is well
known that the separation power of a column increases only with the square root
of the column length, whereas the disadvantages of long columns (high carrier
gas pressure, long dead times and the fact that due to the compressibility of the
carrier gas only part of the column is operated at optimal flow conditions) are
directly related to the column length.
To separate a complex mixture by chromatography, the first requirement is to
achieve a separation performance that is at least as large as the number of com-
ponents in the sample. The separation performance can be characterized by the
number of possible peaks (NPP). Struppe [l] has shown that in the range of a
single carbon number, a maximum of 200 peaks can be completely separated.
But in a real sample this number is significantly smaller because the compounds
are not uniformiy distributed. Consequently, for complex samples, the column
efficiency required to obtain a complete separation will have to exceed that of

232 Chapter 9
present day capability substantially. Optimization of the separation by exchang-

ing a single column for one of a different polarity or by variation of the parame-
ters of a temperature program is also limited in these circumstances since an im-
provement in the resolution of some compounds in the mixture may be accom-
panied by the overlapping of other, previously separated, compounds.
But there is still another problem related to the use of single-column systems.
Even in cases where the performance of a single column is good enough to sepa-
rate two components with adequate concentration levels completely, it may be
impossible to detect a late eluting trace component in a large amount of the early
eluting major component. This is due to the fact that Raoults law is only true for
very dilute solutions. In the following, it will be seen that the use of multi-
column systems can solve problems caused by very complex mixtures as well as
the problems related to the analysis of very different concentration levels in real
samples.
9.1.2 Multi-column systems
There are two general ways of tackling the problems listed above. The first is
to introduce a second dimension of separation leading to a dramatically im-
proved peak capacity (at least for the peaks of interest). The second is to influ-
ence the elution order in such a way that the peaks of interest are eluted at free
places in the chromatogram. That means that a multi-column system allows us
either to find optimal separation conditions by selectivity tuning (described in
Section 9.2) or to reduce the complexity of the sample by the use of column
switching techniques (described in Section 9.3). When using a heart-cutting
technique, the latter is identical to the introduction of a second dimension in
separation.
Today multi-column systems are mainly used for improving separating power,
shortening analysis time and for trace analysis. Some general advantages of
multi-column systems are listed below [ 121:
(a) Increase in resolution: the resolution within a particular group of over-

lapping peaks can be improved either by transferring only this group from
the first column to a second column with higher efficiency or another sta-
tionary phase or by a pre-calculable change of the overall selectivity of
the system.
(b) Shorter analysis time: when a particular analysis of a complex mixture is
to be performed, it is necessary to remove the components that are not of
interest in order to condition the system for the next analysis. By revers-
ing the flow of the pre-column, a fast removal of late eluting components
can be achieved.
Multi-column systems in gas chromatography 233
(c) Extended column and detector life: sample components which may con-
taminate the main column can be prevented from entering the column af-
ter a separation in a pre-column. In the same way, a contamination of
specific detectors can be avoided (e.g. chlorinated solvents in a ECD) by
cutting these components out of the system after the first column.
(d) Decrease in detection limits: overlapping by long-tailing solvent peaks
or major peaks can be avoided by heartcutting.
These techniques also offer enhanced possibilities for qualitative analysis in

two ways. Firstly, the availability of two more or less independent sets of reten-
tion data allows a greater reliability of the identification of individual compo-
nents in complex mixtures by chromatographic means [2]. Secondly, one has to
take into account that the modern hyphenated techniques such as GC-MS and
GC-IR give the most reliable identification only if they are dealing with pure
substances. This means that the preceding chromatographic process should be a
multi-column system to avoid peak overlapping.
Still another problem is the many types of samples which cannot be analyzed
on a single column without a prior clean-up procedure. The use of a multi-
column system, designed as a combination of a pre-column for an on-line sample
clean-up and a second analytical column can be extremely useful in such cases.
Some examples are discussed in Section 9.4.
This chapter provides only a general introduction to the use of multi-column
systems in GC. More detailed information can be found in the papers of Bertsch,
Ettre, Giddings, Guiochon, Huber, Purnell, Sandra, Schomburg and others [3-
111. Some interesting examples for the use of column switching in gas analysis
are also given in Chapter 1 of this book. The reader is referred to the literature
cited at the end of the chapter for a comprehensive coverage of the technique and
its applications.
9.1.3 Multi-column chromatography
9.1.3.I DeJinition
In chromatography, multi-column systems can be assembled in a number of
different ways. This large number of variations has led to some confusion in the
literature about the definition of such multi-dimensional GC (MDGC) systems.
Sometimes systems involving multi-dimensional detection such as the hyphen-
ated techniques have been incorrectly considered to be MDGC, although only a
single column one-dimensional separation is used.
In many papers, definitions of MDGC are given [e.g. 13-15], especially of
two-dimensional GC. This term two-dimensional has its origin in thin-layer
References pp. 266268

234 Chapter 9
chromatography where it has been defined as chromatographic migration in one

direction that is followed by a second development in the perpendicular direction
1161. But today this original definition is not transferable to other chroma-
tographic methods like gas chromatography and is limited to only two spatial
dimensions.
A more general definition has been proposed by Snyder [17] as follows.
Multi-dimensional chromatography is a separation process in which a single
sample is subjected to a sequence of chromatographic separations, each of
which:
- acts upon all or part of the separated components from a previous chroma-
tographic step and
- differs in its relative selectivity and/or capacity.
This definition includes all types of chromatographic separations, HPLC

(high performance liquid chromatography, SFC (supercritical fluid chroma-
tography), TLC (thin layer chromatography), CE (capillary electrophoresis) etc.
Multiple separations, where only the column capacity is changed, are also in-
cluded.
For example, the use of two columns with different polarities coupled in se-
ries can be called MDGC, but the term should not be used if the two columns
differing in selectivity are coupled in parallel and lead to the same detector.
However, in some cases, multi-column systems which are not multi-dimensional
can also have a useful application in chromatography.
In the following section, the numerous variants of MDGC are categorized by
different criteria to provide an overview of the many possibilities of this tech-
nique.
9.1.3.2 Variants of column switching
There are many ways in which a chromatographer can vary a multi-column
system. Depending on the analytical problem and financial requirements, the
system can be optimized for the method of coupling, the operation mode, the
type of column and its working parameters. The various possible ways of cou-
pling columns are listed in Table 9.1 [ 181.
Coupling modes and hardware. Routine analysis requires reproducible condi-

tions. Basic functions, e.g. heartcutting, foreflushing, backflushing, distribution
and intermediate trapping, should be time programmed and controlled by the
microprocessor of the GC. All these requirements can only be fulfilled by on-
line column switching. Off-line methods often work quite well but are hardly
feasible for automated routine analysis [ 191.
TABLE 9.1
VARIANTS OF COLUMN SWITCHING
~ ~~
Criterion Variant
Mode of coupling On-lineloff-line

Seriedparallelheries-parallel
Coupling hardware Mechanical (valves)/pneumatic (valveless)
Operation mode Complete or partial eluate transfer
WiWwithout intermediate trapping
WitWwithout monitor detector after the first column
Type of column Packed, micropacked, capillary
Length, inside diameter
Type (selectivity) and amount (capacity) of stationary phase
Working parameters Sequence of columns (column switching)
Column temperature
- isothermal (TI = T2, T1 # T2)
- temperature programmedisothermal
- temperature programmedtemp. prog.
Variants of flow rates (either in the total system or sequentially
for each column
Generally two distinct forms of on-line coupling have been recognized:
(a) A sample is vaporized in an injector and split into separate fractions. All
of these are simultaneously analyzed on columns with different polarity
connected in parallel [20,21].
(b) A sample is injected into the chromatographic system. After the pre-
column, one or more fractions of the complex mixture are directed
straight to the second column connected in series with or without inter-
mediate trapping. Two or more columns can be combined.
These forms can be obtained by connection with mechanical valves (e.g. ro-
tary, slider, piston or membrane valves [22]) or by a pneumatic switch first de-
scribed by Deans [23] (see Section 9.4).
Especially in valve-switching systems with packed columns, many combina-
tions of these two basic forms (serial + parallel) are found in practice. In capil-
lary column switching, the main technique is serial coupling of two columns.
Because of their high separation power, capillary systems with three and more
columns are unnecessary.
Operation mode of serial coupling. A coupled system can work in the three basic
operational modes: complete/partial eluate transfer, witWwithout intermediate
trapping and with/without monitor detection.

236 Chapter 9
In chromatographs with only packed or capillary columns, the complete eluate

is transferred from one column to the other. Disadvantages of splitting such as
discrimination and loss in detection sensitivity are excluded.
On the other hand, if the coupling piece is only used to permanently split the
eluate of the pre-column to a so called monitor detector and the second col-
umn, two complete sets of retention indices and response data for characteriza-
tion of the sample can be determined.
A coupled system where a part of the eluate is split to waste is used if packed
and capillary columns are combined (pre-column = packed, main column =
capillary) and the eluate will be trapped after the pre-column. In most cases the
split is positioned in the coupling piece between the two columns. The only in-
termediate trap without splitting is the total transfer technique (see Section
9.4.3).
In routine and on-line analysis, the advantages of the direct combination of
columns without an intermediate trap (a simple, inexpensive and easily main-
tained instrument) predominate. In most cases problems caused by merging re-
solved peaks in the second column can be avoided.
Systems with intermediate traps [24,25] are relatively complex and expensive.
By multiple enrichment of components, this method is suited for trace analysis.
The connection of a high capacity packed pre-column to a selective capillary
column is of advantage for many applications.
By refocussing the bands after the pre-column, an exact starting point for each
fraction in the second column is obtained, so that retention data from each col-
umn are available.
If the eliminated components of a mixture are also of interest, the fractions
which have been cut out of the chromatographic system after the pre-column can
be measured by a second detector. The selection of this monitor detector de-
pends on the eliminated substances. In laboratory GCs, such a detector is in-
stalled basically by the use of serial-coupled capillary columns. In on-line pro-
cess GCs, which are applied to only one separation problem, a second detector is
frequently dispensed with because of the cost. Nevertheless, if the cut compo-
nents are of interest, the cut outlet can be connected to the main detector.
Type of colurnn. Many applications are to be found in the literature, either with
packed and micropacked or with capillary columns in multi-column systems.
The variation in length, inside diameter, type and amount of stationary phase
depends on the requirements of the analytical problem. Either gas-solid or gas-
liquid columns can be used and sometimes both together.
In contrast to packed columns, most separation problems can be solved with a
capillary system with a comparatively small number of stationary phases because
of its higher separating power. Except for the separation of permanent gases
which is a domain of the PLOT (porous layer open tubular) columns, WCOT
(wall coated open tubular) columns predominate. SCOT (support coated open
tubular) columns find relatively little use.
Working parameters. The sequence of columns is controlled by mechanical or

pneumatic switching devices and can be changed by the operation software of
the chromatograph. A typical time events program for multi-column-GC soft-
ware is shown in Fig. 9.1.
Laboratory analyses are either isothermal or temperature programmed. Tem-
perature programming is still unusual for process GCs; for an on-line analysis
method like process gas chromatography constant instrument conditions, e.g.
oven temperature, carrier gas pressure or switching events, are extremely impor-
tant for reproducible cyclic runs under rigorous conditions and an oven con-
struction with high thermal mass guarantees reliable isothermal analysis.
In single oven systems, the maximum temperature that can be applied is lim-
ited by the column of least thermal stability. By the use of double oven systems,
TIHE E V E N T S P R O G R A H ChannelA
Command valid for streau n=1

Tm
ie Function Action 1.,4 5..8 9.12 13.. 21.. 25.. 29..
1 0.050 Backf lush on 1111 1110 0000 0000 0000 0000 0000
2 0.300Sample change 1111 1110 0000 0000 0000 0000 0000
3 0.200 Liquid injection on 1111 1110 0000 0000 0000 0000 0000
4 0.500 Liquid injection off 1111 1110 0000 0000 0000 0000 0000
5 0.050 Cut on 1111 1110 0000 0000 0000 0000 0000
6 2.150 cut off 1111 1110 0000 0000 0000 0000 0000
I 2.600 Cut on 1111 1110 0000 0000 0000 0000 0000
8 6,200 Cut off 1111 1110 0000 0000 0000 0000 0000
9 6.200 Backf lush off 1111 1110 0000 0000 0000 0000 0000
10 9.900 Report A 1111 1110 0000 0000 0000 0000 0000
11 9.950 Basic-Exec ANALoG_A 1111 1110 0000 0000 0000 0000 0000
12 9.950 Basic-Exec PRINTW 1111 1110 0000 0000 0000 0000 0000
13 10,000 End of run 1111 1110 0000 0000 0000 0000 0000
Fig. 9.1. Typical time events program of a multi-column process application (time events for the
analysis shown in Fig. 9.7). In process GC, backflush valves are inversely configurated. The valve
position backflush o f f means reversed flow through the precolum, so-called backflush.
Referencespp. 266268
238 Chapter 9
an optimal individual temperature, isothermal or temperature programmed, can

be selected for each column. The advantages of double oven systems are listed
later in Section 9.3.1. Finally, a less obvious use of dual oven systems is multi-
chromatography (see Section 9.2). This technique is based on the possibility of
controlling the carrier gas pressure over the complete system, either before the
pre-column or before the main column.
9.2 SELECTIVITY TUNING IN SERIES-COUPLED COLUMNS
High-resolution gas chromatography implies the optimization of column effi-

ciency, selectivity and capacity ratio. With the introduction of capillary columns,
the main emphasis was laid on their high efficiency. The optimization of the se-
lectivity was not of particular interest. Due to the demands for analyses in very
low concentration ranges it was recognized that many samples were more com-
plex than had been originally thought and the necessity arose for capillary GC
with high selectivity. Such a system can be obtained by selectivity tuning. This
means that the selectivity of a system is adapted to the analytical need by creat-
ing a selectivity between two more or less extreme selectivities. Sandra et al.
[lo] suggested that in capillary GC, selectivity tuning can be accomplished in
one of three ways: by using tailor-made stationary phases, by using mixed
phases, or by serial coupling of columns with different stationary phases. The
last variant is the most elegant method for producing a tuneable selectivity and it
offers the chance of using standardized stationary phases with known retention
properties.
The theory of series-coupled columns is discussed in detail in some papers of
Buys, Ettre and Pumell and their co-workers [5,9,26,27]. We only wish to re-
mind the reader that the retention time of a solute in a system of two coupled
columns (A and B) can be expressed as
where tR is the retention time (system), K is the capacity ratio, L is the column
length and u is the average carrier gas velocity.
From this expression one can derive four principal ways for selectivity tuning:
- variation of K values by changing the stationary phases (chemical nature

and/or film thickness);
- variation of K values by changing the column temperatures;
- variation of the column lengths (segmented column method);
- variation of the average carrier gas velocities of the columns (multi-
chromatography).
It is quite clear that the use of other stationary phases or the variation of the
column lengths in a system consisting of very different single columns will lead
to dramatic changes in the selectivity of the system. However, the disadvantage
of these two methods is that a new column system is necessary for an optimiza-
tion procedure after every optimization step. This would be a very time- and
material-consuming process which would only be useful for the development of
a routine analytical method.
For the optimization of frequently changing analytical problems, selectivity
tuning by variation of column temperatures or flow rates is more attractive. Gen-
erally, one has to consider that the limits for selectivity tuning are given by the
selectivities of the individual columns. That means that small differences in the
selectivity of the individual columns allow only a small range for selectivity
tuning. Therefore, it is useful to combine columns which are as different as pos-
sible in their retention characteristics. To demonstrate the effect of selectivity
tuning by variation of temperature even for very similar compounds, we show in
Fig. 9.2 the dependence of the retention indices of the 15 octane isomers listed in
Table 9.2 on the temperature of the first column (for more details see [28]). The
temperature of the second column was kept constant. In this case, not only dif-
TABLE 9.2
COMPOSITION OF THE TEST MIXTURE FOR SELECTIVITY TUNING
Peak Component
1 n-Hexane
2 2,2,4-Trimethylpentane
3 n-Heptane
4 2,2-Dimethylhexane
5 2,5-Dimethyihexane
9 2 3,3-Trimethylpentane
11 2-Methyl-3-ethylpentane
12 2-Methylheptane
13 4-Methylheptane
15 3-Methyl3-ethylheptane
16 3-Methylheptane
17 3-Ethylhexane
18 n-Octane
GC-conditions: instrument; Sichromat 2 (SIEMENS); columns (A) 50 m X 0.32 m m i d . OV-1, fs;

(B) 1.5 m X 1 mm i.d. graphitized carbon black (micropacked), ss; carrier gas, 4.3/4.0 bar hydro-
gen.

240 Chapter 9
1s :16Q'C
800 18
'M
12
m
n
17
75c
7
3
70(
650
60 70 80
'A fcl
Fig. 9.2. Selectivity tuning by variation of temperature (for substances, see Table 9.2). GC condi-
tions: column A, 50 m x 0.32 mm OV-1; column B, 1.5 m X 1 m micropacked graphitized thermal
carbon black (GTCB), carrier hydrogen, temperature of column B, 160C.
ferent polarities but also a combination of two different chromatographic

mechanisms (partition and adsorption) were used to optimize the separation. It
can be seen that for selectivity tuning by variation of column temperatures, a
double oven chromatograph or at least two chromatographs with a connection
line are necessary to operate both columns independently at optimum tempera-
ture. The advantages of a double oven chromatograph are discussed in more de-
tail in Section 9.3.1.
Multi-column systems in gas chromatography 24 1
Variation of flow-rates in a system of series coupled columns (see Kaiser and

co-workers [29]) is a very special way of selectivity tuning. In practice, carrier
gas is added at the connection point between the first and second column
(leading to a higher flow-rate in the second column and an increasing influence
of the selectivity of the first column on the system selectivity), or carrier gas and
consequently a part of the sample, are vented at the connection point (leading to
an increasing influence of the second column on the system selectivity). With
this method, impressive changes in the system selectivity can be achieved with a
very simple apparatus; the selectivity can also be easily pre-calculated. The
problem is that at least one column is not operated at the optimum flow rate.
This means that the range of possible flow rates is limited by the effect of a re-
duced number of theoretical plates according to the van Deemter equation. Con-
sequently, the use of hydrogen as carrier gas for selectivity tuning is recom-
mended because of its broader van Deemter optimum.
9.3 COLUMN SWITCHING TECHNIQUES
9.3.1 General
Column switching was first developed for process control [32]. Some indus-
trial processes require the analysis of only a single compound or a limited group
of substances which may be in complex matrices or emerge after main compo-
nents which are not of analytical interest. Under these circumstances, it is desir-
able to develop a chromatographic system which allows the fractionation of the
sample and the removal of fractions of no interest. Analyses using temperature
programming in laboratory gas chromatography, especially the removal of high
boiling compounds from the system, can be efficiently solved in an isothermal
mode by the use of a column switching system. One of the first applications of a
column switching system in process control was published by Villalobos and co-
workers [30] in 1961. Since then, the advantages of column switching have also
been used in laboratory applications. Figure 9.3 shows in a simplified form, what
effects can be achieved by the application of the special column switching tech-
niques, described later. Figures 9.4 and 9.5 illustrate the schematic flow paths of
typical column switching systems for valveless as well as for valve switching
systems.
A very helpful tool for the development of column-switching methods is
the availability of two independently controlled ovens. Only a few manufac-
turers offer double oven chromatographs for laboratory analysis as well as for
process-control. The advantages of such instruments can be described as fol-
lows:

242 Chapter 9
Removal of high baling mpou&,

fast reconditioningof the system
Reduchon of compkx samples

Shortening of analysts hme onto onty a few compounds
of i n t d
r
Protection of cdurnns or
Anal- of very dtfferenI types of detedm from stressing
compounds in only one system subdances
HeartCut
Decrease in Wecbon limtts Analysrs of traces in the

tailing of main components
Vanatm of retmllon timcw for

compounds of interest
Fig. 9.3. Advantages of the use of different column switching techniques.
- The overall temperature limit of a chromatographic system is no longer

determined by the phase with the lowest temperature limits; stationary
phases with high and low thermal stability can be operated independently.
- Gas-solid chromatography usually requires higher separation temperatures
than gas-liquid chromatography; in a double oven system, the combina-
tion of both separation principles is possible in such a way that both col-
umns can be operated at their optimum temperature.
- It is possible to adjust the selectivities of both columns by tuning their
temperature (see Section 9.3) independently, thus having a very elegant
way of combining the advantages of selectivity tuning and column
switching.
straieht on
Fig. 9.4. Column switching using valves.
NV 2
Straight on cut Backflush
Fig. 9.5. Column switching using a live-switching system.

244 Chapter 9
- When combining two identical columns, the first operated in a tempera-

ture programmed and the second in an isothermal mode, one can exploit
the second column as an identification column for the determination of
highly precise isothermal retention indices and in this way the advantages
of temperature programmed separation can be combined with those of
isothermal identification of the separated components [ 181.
Finally, the development of the modern microprocessor controlled instru-

ments incorporating a digital adjustment and control of temperature and even
pressures and flow rates allows very precise and reproducible results and the
built-in time programmable functions and integration methods make column
switching systems relatively easy to use.
9.3.2 Backflushing
Backflushing is one of the most important switching steps and it is used in

almost all process chromatographic applications. The aim of this technique is to
shorten analysis time and to obtain a definite end to the analysis. Analysis time
can be reduced by eliminating the compounds which are eluted on the first col-
umn after the last compound to be measured. After the compounds of interest
have moved from column A to column B and the less important components are
still in column A, the direction of the carrier gas stream in column A is reversed
(see Figs. 9.4 and 9.5). This means that all components that have not left column
A at the time of switching are flushed out from the separation system. A very
typical example of the reduction of analysis time by the use of backflush is the
determination of traces of acetylene in ethylene. Figure 9.6 shows the chroma-
togram of such an analysis without backflush. From the bottom part of the fig-
ure, it can be seen that the use of backflush allows more than twelve analysis
cycles in the same time. Another advantage of the backflush technique is a
definite end to the analysis. In contrast to laboratory gas chromatographs where
late eluting components are usually removed from the system by temperature
programming, the backflush technique removes these components without ther-
mal stress to the columns. Moreover, it is the only method which can guarantee
that all components that can be eluted are really flushed out of the system so that
late eluting components are not disturbing further analysis sequences as unex-
pected peaks or as a baseline drift. The column length ratios can, usually without
any problem, be arranged in such a way that the elution time for the last com-
pound of interest in column B or B and C is at least as long as the elution time of
that compound in column A. Another common trick when using backflushing is
to adjust the backflush gas flow rate to be greater than the forward flow rate.
Fig. 9.6. Shortening of analysis time by the use of backflush, analysis of trace amounts of acetylene
(1) in ethylene (2).
That means that the time for backflush is long enough to remove all disturbing
compounds from the system and immediately after the elution of the last peak of
interest a new analysis sequence can be started.
9.3.3 Cutting
Section 9.3.2 described how the use of backflushing can avoid interferences
or disturbances of the analysis by compounds that would be eluted after the last
component of interest. In addition, the cut technique allows venting out of the
chromatographic system components eluting in front of, or between components
of interest. This can be done by a time-programmed switching of the gas flow
after one or more pre-columns either in the straight-on position (components are
transferred to the main column and to the detector) or in the cut position
(components are vented out of the system through a needle valve or a capillary
restrictor with the same flow resistance as the main column). The flow-paths for
both cases are shown in Figs. 9.4 and 9.5, respectively. There are three important
aims in the application of the cut technique:

246 Chapter 9
- Extended column life and prevention of detector contamination by selec-

tive transfer of only the analytically significant components into the main
column. We have pointed out the advantages earlier.
- Reduction of the complexity of multi-component samples and thereby re-
duction of the probability of peak interferences. This is a very helpful tool
when only a few components of a complex sample are to be determined.
In cases where different stationary phases are used the well known effect
of interferences between components which are separated on the individ-
ual columns but have the same overall retention time on the combined
columns can be avoided.
- A third and very important application of cutting techniques is the so-
called heart-cutting. In trace analysis, the column usually has to be
overloaded in order to obtain a sufficient signal from the trace component.
This causes difficulties when the trace is eluted on the tail of the major
component. Using a cutting device so that only the trace and the underly-
ing small amount of the major components are transferred to the second
column gives a substantial improvement in separation and a decrease in
the detection limit for the trace component. The second column only has
to separate the trace from the small amount of the tail of the major com-
ponent. The typical peak shape of the tail can be seen in the simple ex-
ample of a heart cut given in Fig. 9.7.
9.3.4 Distribution cutting
The column switching configuration called distribution cutting is used either

to shorten analysis time or in cases where very different components are to be
separated on two different columns simultaneously. It is realized by the same
switching configuration as for cutting but in distribution mode, the cut compo-
nents which are vented to waste in cut-mode are by-passed to the detector
through a needle valve or a capillary restrictor only or to a combination of an
additional column and a restrictor which together have the same flow resistance
as the main column. A typical example of the shortening of analysis time is
given in Fig. 9.8. The second column is only necessary for the separation of
components 11-13. Components 1-10 will be by-passed directly to the detector
after the first column.
Figure 9.9 shows that the elution of all the components through the second
column would lengthen the analysis time. This is significantly reduced by the
use of the distribution technique. A typical example of the other strategy of dis-
tribution, the simultaneous separation of very different components within only
one analysis is the application of permanent gases and hydrocarbons. A pre-
column has to separate the sample into a hydrocarbon fraction and a permanent
gas fraction. The permanent gases are separated on molecular sieve whereas the
hydrocarbon fraction is distributed to another column which would not be able to
separate the permanent gases. An example of the resulting chromatograms is
given in Fig. 9.10.
Fig. 9.7. Improved detectability of traces in the tail of the major component by heart-cut. 1, Main
component; 2, 3, trace components to be determined; 4, typical peak shape of the cut. Top, without
heart-cut; bottom, with heart-cut.

248 Chapter 9
Fig. 9.8. Shortening of analysis time by distribution (chromatogram), analysis of a gasoline frac-
tion, GC conditions: column A, 17 m X 0.20 mm HP-PONA; column B, 33 m X 0.20 mm HP-
PONA, carrier hydrogen, temperature 30C. 1, butane; 2, i-pentane; 3, n-pentane; 4, 2,2-
dimethylbutane; 5 , 3-methylpentane; 6, hexane; 7, methylcyclopentane; 8, 2,4-dimethylpentane; 9,
benzene; 10, cyclohexane; 1 1, cyclopentane; 12, 2,3-dimethylbutane; 13, 2-methylpentane. Sepa-
ration of components 1-10 on column A only and distribution directly to the detector, separation of
the components 11-13 on columns A and B.
Retention Imin.1
25
20 - ----
-~
~
i ~-
15
10
5
0 - --- _- 7
1 2 3 4 5 6 7 8 9 1 0 1 1 1 2 1 3
Measuring components
t(R) precolumn t(R) main column '/ minimal cycle 1

Comp.11-13 not separated after precolumn
Fig. 9.9. Comparison of retention times with and without distribution (components and GC condi-
tions see Fig. 9.8).
A
1
5
2
Fig. 9.10. Distribution onto two different types of columns, separation of permanent gases on a
molecular sieve (A) and of lower hydrocarbons on a modified Spherosil (B). 1, oxygen; 2, meth-
ane; 3, ethane; 4, ethene; 5, propane; 6, ethine; 7, iso-butane; 8, n-butane; 9, propene.
9.3.5 Special switching techniques
9.3.5.1 Summation of backjlushed compounds

For certain applications, it is required that all components, even very high
boiling ones must be detected. A straight elution of all components often pro-
longs analysis time unnecessarily, and in many cases, higher boiling compounds
do not elute at all, causing build up on the column with deterioration of effi-
ciency or selectivity. In these cases and when only the determination of late
eluting compounds as a sum parameter is of interest, a backflush sum is used.
Backflush sum means in practice that the same switching configuration as for
backflush is used but in this case, the backflushed effluent is not flushed out of
the system but is by-passed to the detector. All components which, at the time of

250 Chapter 9
switching, are still in the first column, are backflushed. In this manner, the sepa-
ration already achieved is eliminated by the reversal of carrier gas flow; all com-
ponents are eluted simultaneously and detected as one peak.
9.3.5.2 Stoppedflow/stuttering
In all systems with more than one flow path to the detector, it is possible that
different components are eluted to the detector by both paths simultaneously. If
there is no possibility of avoiding this by a proper arrangement of column
lengths and flow rates, then two more or less tricky column switching techniques
can help to solve the problem without the necessity of using a second detector.
The first variant (unusual in capillary systems) is to stop the flow in one column
for a short time by by-passing the carrier gas through a needle valve with the
same flow resistance as the stopped column. In this way, all components
which are in the stopped column will elute a little later. But in every case, one
has to take into account that during the time the flow is stopped, peak broadening
processes by diffusion continue. Our experience is that immobilization of some
components, especially hydrogen, often results in band broadening or a loss of
sensitivity.
The same effect, a selective delay of one or some components, can be
achieved by short periods of flow reversal. Usually the backflush mode can be
used for this technique. Switching of the backflush valve for a short time causes
a delay for all components which are still in the first column. This so-called
stuttering technique can also help to avoid the simultaneous detection of two
separated components from different flow paths, but of course the application of
stuttering is limited by the same disadvantages as stopped flow.
9.3.5.3Recycle Chromatography
The idea in recycle chromatography is to achieve the required separation, not
by the use of a very long column, but by a repeated separation of the sample on
one and the same short column. This can also be realized with a column switch-
ing technique. Recycle chromatography offers the chance to get a large number
of theoretical plates with a very small carrier gas pressure and, in the case of
capillary columns, with short and inexpensive columns. The principles of recycle
chromatography especially in preparative mode have been treated by Chizkov et
al. [31]. Some of the known applications are listed in [4] but it seems that the
technique has not found widespread use up to now.
9.4 PRACTICAL ASPECTS
Instrumentation for multi-column gas chromatography ranges from very ele-

mentary arrangements to highly complex and automated systems. In the simplest
case, only a conventional gas chromatograph with single inlet, single detector
and two columns is required. These two columns can be coupled either by valves
or more elegantly by a valveless pneumatic switching device. Instruments spe-
cially designed for multi-dimensional GC may also incorporate such functional
elements as auxiliary inlets/outlets, selective detectors, dual ovens, automatic
timers and various intermediate traps for focussing components of interest. A
multi-column system using traps is one possibility for the utilization of column-
switching as a sampling technique.
9.4.1 Valve switching
Multi-column systems using valve switching have been in use successfully for
more than 30 years [32]. Valves are the simplest way of switching effluent from
one column to another and are very similar to gas sampling valves. Others used
for column switching are rotary, sliding or piston valves. Some versions are de-
signed with up to 12 ports, miniaturized or remote controlled. Figure 9.4 (see
Section 9.3.1) shows a diagram of a complex valve switching system.
Commonly, valve switching systems are used with packed columns. However,
in recent years, packed columns have been more and more displaced by capillary
columns, especially in single but also in multi-column systems, because of their
greater efficiency. These capillary multi-column systems are mostly switched by
pneumatic switching devices. Nevertheless, for some analytical requirements,
the use of mechanical switching in combination with packed columns has some
advantages. As many different column configurations as necessary can be built
and therefore many analytical problems can be solved with one system. Their
relatively large internal volume does not affect the analytical separation because
of the high carrier gas flow rates through packed columns. However, if micro-
packed or capillary columns with smaller inside diameter and lower operating
flow rates are used, peak broadening and tailing with a loss of efficiency may
occur. These adverse effects can be overcome by cold trapping the solutes on the
head of the column. The temperature limitation due to the sealing material
within the valve is another disadvantage when using mechanical column
switching by valves. None of the materials inside the valves present an ideal sur-
face for gas chromatography. Adsorption and memory effects can limit the ap-
plication, especially for trace component analysis.
However, because packed systems with valve switching are simple, inexpen-
sive, versatile and easily maintained, such systems are still used, especially in
on-line process control and for gas analysis. A very simple but reliable and rapid
analysis for process control is the determination of sulphur components in the
Claus process shown in Fig. 9.1 1.
Refirences pp. 266268

252 Chapter 9
i min. 6
J
2
J
Fig, 9.1 1. Rapid determination of sulphur components for Claus-process control. 1, nitrogen;, 2,
carbon dioxide; 3, hydrogen sulfide; 4, carbonoxy sulfide; 5, sulphur dioxide; 6, water.
9.4.2 Valveless flow switching
A more complex method of redirecting flows in chromatographic systems op-

erating at elevated temperatures is the use of pneumatic switching. In contrast to
mechanical switching, all moving parts are located outside the heated zones and
the separation path. In such a pneumatic system, the demand for a zero dead vol-
ume column coupling is obtained by different types of coupling devices mostly
made of glass or noble metals. Consequently none of the problems of leaks, tem-
perature limitations, adsorption and memory effects or loss in efficiency associ-
ated with mechanical valves apply to such a system.
FID 1
Aromatics
FID 2
Saturates
and Olefines
,I ,p ,*.
-I
,I,, Y YI . . . I I-
C
I"
I
." .
.I- .>I.
d-
-3
I
II". "I m
C
.-.,
I
I 111," .. -
C I u
.,.
, --s
Siemens SiCHROMAT 2
Fig. 9.12. Determination of individual components (paraffins, olefins, naphthenes and aromatics) in
hydrocarbons mixtures on capillary columns. 1, separation of aromatics (xylene and higher) on a
polar pre-column and distribution to FID 1; 2, transfer of lower to a non-polar second column and
separation up to C9, benzene and toluene, detection of saturates and olefins by splitting of eluent;
3, detection of saturates only on FID 3 by means of scrubbing out the olefins.
PONA analysis (paraffins, olefins, naphthenes and aromatics), a typical

analysis in the petroleum industry, demonstrates the high performance of a
valveless switching system using capillary columns (Fig. 9.12).
One critical component in pneumatic switching is the carrier gas supply con-
trolled by pressure regulators. The pressure regulation should be very repro-
ducible under all conditions. Two typical valveless flow switching systems are
described below.
9.4.2.I Deans-switching
The first pneumatic switching system was developed by Deans [23]. A sche-
matic flow path of such a system is given in Fig. 9.13. The principle is based on
254 Chapter 9
solenoid operation
ON/OFF ~ valve
manual operation
ON/OFF ~valve
Fig. 9.13. Flow diagram of a heart cutting system for two packed columns due to Deans [33].
the balance of flows by in-line restrictors and two pressure controllers (head and
medium pressure) indicated on gauges (PA, P,). By opening and closing external
valves (SVl), the direction of flow can be changed in this two-column system.
Before starting analysis, the following flow adjustments should be carried out.
Basic adjustments
- Adjustment of head pressure PA to the optimum flow with PR1.
- Adjustment of medium pressure P, by isolating PR2 with the odoff valve
M V , so that the naturally occurring pressure can be read on P, . For this
operation SV 1 is closed.
- R1 is chosen such that a small fraction of pre-column effluent is passed to
the monitor detector (Dmn).
- NV2 should approximately balance the restriction of the main column.
Two phases can be distinguished in the operation modes of the illustrated

system:
Sample transfer. Effluent eluting fiom pre-column A enters main column B in

the direction towards the main detector D ~ nSV1. is closed during the operation.
A small fraction of effluent is split to monitor detector DmOn.PR2 supplies the
junction of the columns with a small amount of carrier gas.
Heart cutting. Effluent eluting from pre-column A is vented out of the chroma-
tographic system through needle valve NV2. SVl is now open. The gas and the
effluent coming from the pre-column flow out to the atmosphere.
In addition, a small amount of gas coming from PR2 ensures that a clean cut is
obtained.
By modifling the illustrated system with an additional valve after PRl, the
backflush mode can also be realized.
This system has the following advantages. Transfer times between columns
can be extremely short (milliseconds) and this makes very sharp cuts possible. A
fraction or fractions can be transferred from the first column to the second with-
out interference from components of no interest, which are vented.
However, the use of Deans-switching is limited by the following:
- Only a few switching modes are possible (backflush and heartcut).

- Retention time shifting on the first column is the result of high pressure
differences in the coupling piece when switching between cut and transfer
mode. This affects the cut time period.
- The system is mainly applicable to packed columns. The use of capillary
columns requires a focusing trap for the second column.
- The cut valve is still in the sample path. This means a limited lifetime for
this valve.
- The flow through the cut exit depends on the switching position and thus
a constant flow is not available at this point. Therefore monitor detection
is only possible when using an additional restriction outlet between cou-
pling piece and cut valve. With this modification, the effluent is continu-
ously split to the monitor detector.
Deans demonstrated that, in some cases, two packed columns in series with
different stationary phases and heart cutting between them can have a much
greater separation power than a single capillary column [33]. In such a packed
column system, the source of separation power for complex mixtures, in spite of
lower number of plates, lies in the difference in selectivity between the two sta-
tionary phases and in the elimination of interfering components of no interest.
However, packed columns were mainly used in coupled systems for simple ana-
lytical problems having a less complex sample composition.
9.4.2.2Live-switching
The live column switching system [34] is based on the principle of pneumatic
switching developed by Deans. The differences to Deans-switching are the fol-
lowing:

Chapter 9
Fig. 9.14. Scheme of the small pressure differences in the coupling piece.
- In contrast to the Deans system which operates with significant flow dif-
ferences between the pre-column and main column, the live system is
based on small pressure differences (see Fig. 9.14) in the coupling piece,
which results in very small flow differences. Therefore, the live system
guarantees a constant flow through all the columns and restrictors, so that
the cut exit can also be connected to the detector.
- The live switch consists of a special coupling T-piece which is a part of a
"pneumatic bridge" configuration (see Fig. 9.5). It operates in a very
similar manner to the Wheatstone bridge, consisting of four (flow) resis-
tors (needle valves NV2, N V 3 , and restrictors R1, R2). The differential
pressure (6P) in the bridge diagonal is adjustable in the negative direction
with N V 2 and instantly switchable with solenoid valve SV CUT.
- There is an additional flow channel with only a small resistance, which is
located in one diagonal. Either the total flow from the first column is di-
rected into the second column or it is diverted from the first column via
the restrictor R1 into the cut exit, due to a reverse flow depending on the
differential pressure applied.
- The specially designed T-piece, shown in Fig. 9.15, avoids unswept vol-
umes by small gaps - especially between the column and the coupling
tube. Back diffusion of the sample into the gaps is thereby eliminated and
1-
Fig. 9.15. Dead-volume free live-T-piece for coupling two capillary columns.
peak distortion is also prevented in the cut path. Capillary columns can be
used as well as packed columns without loss of efficiency.
- More principal column configurations are possible (backflush, heartcut,
distribution and backflush sum).
In this system, high precise pressure controllers are used, so that the relation
between head and medium pressure is constant over the complete analysis time.
Only under correct pressure regulation are the switching times of, e.g. heartcut,
precise and reproducible. Regulators for this system based on the principle of a
nozzle/baffle plate arrangement are built up completely without a diaphragm, so
that the carrier gas supply cannot be contaminated by degassing softening agents
and also the influence of ambient temperature change is eliminated.
From the above characteristics, the advantages of live-switching are clear. In
Fig. 9.16, this system is compared with a mechanical column switching unit
where the switching valves are located in the heated zone.
For the first time, this technique made it possible to use capillary columns in
process chromatography [35], where it is essential to switch columns (the mini-
mum requirement is the possibility of backflushing the column). Figure 9.17
258 Chapter 9
Prevention of band broadening

and memory effects
Inertness
Speed of switching
cost
Ease of installation
Ease of operation
Avai Iabi I1t y
Suitability for automation,
Capillary GC
and High Temp. GC
unsuitable moderate good excellent
I -valves valveless 1
refers to poctcd EOIUEIRS d y ;
for CtpilLry colrtcnos valves arc not likely to bc asable
Fig. 9.16. Comparison of a valveless live system with a mechanical column switching system.
shows a typical process GC application (by packed and capillary techniques) in a

steam cracker plant for process control. The higher performance of capillary col-
umns in contrast to packed columns is demonstrated. The capillaries are coupled
without valves, the packed column system is switched with valves.
9.4.3 Column-switching as a sampling technique
If trace components are to be determined in complex matrices, special prob-

lems arise using single column GC systems.
The signal-to-noise ratio of trace components is normally too low for reliable
detection. The significant components must be enriched and transferred from an
unsuitable matrix into a new solvent matrix. After sample pre-treatment by, for
example, extraction or LC separation, a large sampling volume must be intro-
duced into the chromatographic system to increase the signal-to-noise ratio of
the trace components. The consequent overloading results in poorer resolution
and the broad solvent peak can overlap neighbouring minor components.
Another limitation is the possible disturbing influence which a major compo-
nent may have on the performance of the column (e.g. water using molecular
sieves) and detector (e.g. chlorinated solvents using ECD).
0.0 16.0
J- 8.0
Fig. 9.17. Separation of a C1<4 hydrocarbon mixture. I , i-butane; 2, n-butane; 3, 1-butene; 4, i-

butene; 5, trans-butene; 6, cis-butene. (a) With packed columns in 16 min; (b) with capillary col-
umns in 8 min.
Integrated pre-separations, like those performed on-line within the closed

GC system, are preferable to off-line sample pre-treatments which carry the risk
of undesirable changes in the original relative concentrations of a significant
component.
In the following sampling techniques, the first using a two-dimensional capil-
lary column system and the second, using a system with different type of col-
umns (packed-capillary) and intermediate trapping, are discussed.

260 Chapter 9
9.4.3.I Samplingfr.om a capillary pre-column to a main capillary column

Grob Jr. [36], Schomburg et al. [37] and others pointed out that by column
switching in a typical MDGC system containing two or more columns coupled in
series, the problems mentioned above can be avoided. In this system, in which
the first column serves as a sampling unit for the main column, only selected
parts of the original sample are, in most cases, transferred into the second col-
umn. This operation can be considered a selective sampling technique, if the
analytical data (response, retention) are taken from the separation in the second
column.
Other selective sampling methods like pneumatic sampling switching tech-
niques (Deans- or live-sampling [ 3 S ] ) , temperature programmed sample transfer
(e.g. moving needle [39]) or head-space analysis [40]) may also be useful for
overcoming the negative influence of column overloading by the matrix solvent,
but are not discussed here.
Three examples of multi-dimensional sampling onto capillary columns are
discussed in more detail:
(a) Polarhon-polar column-coupling. The aim of the analysis is a simulta-
I
neous determination of polar substances by monitor detection and non-polar hy-
drocarbons transferred to the main column and the main detector. The determi-
I 1
C-Heranr Bulanol 2
/I
1 1
2 N-Hexane
10
11 Isobutanol
12 Bulanol
13 Benzene
5 Acelone J 1 4 Toluene
15 Ethylbenzene
16 P-Xylene ,
1 7 M-Xylene
, 18 0-Xylene
o?;
I
1 Light benzene
I !R
.. .
Stenens SiCHROMAT 2
I 5
- lime. minutes
5 I
Fig. 9.18. Multi-dimensional separation of oxygenated components in gasoline using a system of

coupled columns with different polarities. (A) Chromatogram of gasoline obtained with polar pre-
column (TCEP); (B) separation of cuts on a non-polar main column (OV-I).
nation of oxygenated components in gasoline (Fig. 9.18) demonstrates the im-

provement in resolution for significant groups of peaks by changing polarity
from the first to the second column. The first separation is carried out by using a
very polar tris-(cyanoeth0xy)propane (TCEP) column. All aliphatic components
up to C t t are eluted before the aromatics. The unresolved components are sepa-
rated on the main column after the first column. Overall analysis time is 35 min.
(b) Coupling of columns with identical polarity but difSerent film thickness.
The separation of a crude oil fraction in the C number range C4-C30 seen in Fig.
9.19 is a remarkable example of the separation of mixtures ranging widely in
volatility. The first separation is carried out by using a non-polar thin film col-
umn. All components, covering the entire volatility range, are eluted within a
reasonable time. The second separation is to resolve the volatile hydrocarbon
isomers of the C number range up to CI2,now using a non-polar thick film col-
umn. Optimal operation of the two columns is only achieved with different tem-
peratures or with temperature programs maintained within two ovens. The
Fig. 9.19. Separation of crude oil using a system of two coupled alkylpolysiloxane columns with
different phase ratio. (A) Chromatogram of crude oil. (B) Crude oil pre-separation. Cut after elu-
tion of dodecane. (C) Separation of cut in carbon number range C2-C12. Sample, 0.3 pl of crude
oil; columns (A,B), 30 m SE 52, 1 pm film thickness; 50 m OV-1; temperatures (A,B), 5 min 50C
iso, 50-300C (8 "Chin), (C) 5 min iso, 0-50C (lO"C/min), 50-250C (CC/min); carrier gas
(A,B), 1.1 bar H2, (c) 0.8 bar H2; analysis time, 40 min.

262 Chapter 9
overall analysis time, if the separations are performed simultaneously, is no

longer than 40 min with a greatly increased peak capacity.
(c) Coupling of identical polar columns for heart cutting. The determination
of low concentrations of components in the tail of the major component is the
aim of the analysis (see Fig. 9.7). More details about this technique of sampling
by heart-cutting are given in Section 9.3.3.
9.4.3.2 Coupling of columns of digerent type with intermediate trapping
A switching system consisting of capillaries has disadvantages; because of
their low sample capacity, the sample is often split in the injector and, therefore,
the amount of sample may be too small to get good results in trace analysis. To
achieve an improvement in the lower limit of detection, it is desirable to transfer
the total amount of trace components accumulated within the pre-column, with-
out splitting, into the main column. However, interference by the large amount
of solvents and other undesired by-products has to be avoided by means of flow
switching, e.g. by backflushing or cutting, after the pre-column.
By coupling a packed pre-column with a large sample capacity with a high
resolution capillary main column, these problems do not arise [41,42].
Because the carrier gas flow through a packed column is much higher than
through a capillary column, only a part of the sample can be transferred. There-
fore, the split point is brought from the injector to the union tee between the two
columns. Discrimination effects typical for split injection are thus avoided.
For optimum performance, broad peaks eluting from the pre-column must be
refocused by a cold trap before starting the capillary separation. This trap de-
signed as a connecting tube with low mass for rapid heating can be empty, wall-
coated, or filled with packing material. After one or more pre-separations, the
enriched trace components, condensed in the cold trap, are thermally displaced.
To retain full column efficiency for the separation of the species of interest, the
trapped fraction is ejected into the hot capillary column with a high heating rate
and broad or distorted peaks are eliminated.
If the main separation is performed isothermally, retention index identifica-
tion with high reproducibility is possible.
A modification of the conventional intermediate trap the so-called total
transfer system [43] has the advantages of intermediate traps and also elimi-
nates the problems of splitting after the packed pre-column. Therefore, the total
transfer technique is ideal for sub-trace analysis.
Figure 9.20 shows a schematic diagram of this system. The coupling of the
packed and the capillary column is by a glass tube which is also the trap. Around
the tube, there is heating wire for heating the trap rapidly and transferring the
components to the capillary column. The trap can be cooled by a second glass
tube which is designed as a mantle around the trap. The negative temperature
Denar
Fig. 9.20. Intermediate trapping by total eluate transfer, coupling of a packed with a capillary col-
umn.
gradient when cooling with nitrogen is provided by the inlet of the cooling me-
dium on the capillary side which guarantees a good focusing of the peak.
9.5 STRATEGIES FOR THE APPLICATION OF MULTI-COLUMN

SYSTEMS
9.5.1 General remarks
The full analytical potential of a chromatographic separation can only be used

after the development of a sample specific solution for a particular problem. Be-
cause of the large number of parameters (e.g. injection procedure, type, length
and diameter of columns, types and amounts of stationary phases, column tem-
perature and temperature program, type and flow rate of the carrier gas and type
and working conditions of the detectors), it is unlikely that there will be only one
clearly defined optimal solution for a given problem. There are rather several
possible solutions for every problem to be solved by chromatography, and the

264 Chapter 9
choice depends on the experience of the user, the equipment available and the
effort required for the maintenance of the equipment.
Therefore, it is of special importance to establish a consensus between the
applications engineer and the user about the analytical aims to be attained before
identical / similar
analytical problems
single-column
system
Fig. 9.2 1. Guidelines for the decision to use a multi-column system.

--;~x;x;x;x~x;x;x; Valve Column Switching
. I .
, . Pneumatical Column Switching

, .x I : x . x : 1: x : x . x :
Gaseous Sample
Liquid Sample
Liquified Gas
, . ' 0 Sample with Disturbhg
High Boiling Components
or
Sample with High
Concentralion Differences
Measuring of Trace Components
Multi Component Analysis

Measuring of Key Components
Multi Component Analysis

Measuring of All Components
Sample with a Small Difference

-. ..
of Boiling Point
but a High Difference tn Polarity
. . . . . . . Sample with a Small Differences
. . . . . . .
. . . . .x.x. in Polanty but a
. . . . . . . High Differenceof Bolllng Pcint
. . . . . . ..
Sample wth a Small hfference
i:x: : : : : : :x: of BoilingPoint and in Pdanty
. . . . . . .
y
1:x:
. . . . . . .
, : : : :
. . . . . . .
. . . . . . .
:x:
Sample wth a High Difference
of Bollrng Pant and in Polany
Sample Consisting of
Permanent Gases and
Ltght Hydrocarbons
Short Analysis Times
ChemicallySensitive Substancas
D
.. .. .. .. . .
Ultra Trace Analysis
266 Chapter 9
development is started. The demand for only a single additional component to be

measured, an improvement in the detection limit or a shorter analysis time can
make an existing solution useless.
For this reason, the following guidelines for an applicational development are
kept very general and they cannot show the appropriate solution in every case.
However, the authors hope that for the majority of analytical problems to be
solved by gas-chromatography, a convenient solution can be found with their
help.
9.5.2 Guidelines for the use of single- or multi-column systems
The basic question for the selection of a GC system (single- or multi-column)

is, what kind of analytical problems are to be solved. This is important if the
system is mainly used to analyse identical or nearly identical samples, typically
for process applications. On the other hand, for constantly changing analytical
problems, an adaptation of the system is required, typically for GCs in research
laboratories. Figure 9.21 should help the decision as to whether a multi-column
approach seems to be useful for an individual application.
After the decision in favour of a multi-column solution, the separation and
switching systems should be selected; Fig. 9.22 can be helpful in this. As a rule,
a current analytical problem will comply with some of the characteristics listed.
The proposed technique depends on the frequency of the marked sections. Nev-
ertheless, some individual demands on the system are not taken into considera-
tion, e.g. cost. Also the ease of installation and operation plays an important part
in the selection of the analytical hardware. As a matter of principle, the simplest
possible system should be selected that fulfills the requirements demanded.
9.6 REFERENCES
1 H.-G. Struppe, in: Handbuch der Gaschromatographie, Akademische Verlagsgesellschaft

Geest & Portig (1984), p. 325.
2 T. Maurer, W. Engewald and A. Steinbom, J. Chromatogr., 517 (1990) 77.
3 I.L. Davies, K.E. Markides, M.L. Lee, M.W. Raynor and K.D. Bartle, J. High Resolut. Chro-
matogr. Chromatogr. Commun., 12 (1989) 193.
4 W. Bertsch, Multidimensional Gas Chromatography, in: Multidimensional Chromatography
(H.J. Cortes, Ed.), Marcel Dekker, New York (1990), p. 74.
5 J.V. Hinshaw and L.S. Ettre, Chrornatographia 21 (1986) 561,669.
6 J.C. Giddings, J. High Resolut. Chrornatogr. Chromatogr. Commun., 10 (1987) 3 19.
7 G. Guiochon and C.L. Guillemin, in: Quantitative Gas Chromatography for Laboratory Analy-
ses and On-Line Process Control, Elsevier, Amsterdam (1988).
8 J.F.K. Huber, E. Kenndler, W. Nyiry and M. Oreans, J. Chromatogr., 247 (1982) 21 1,
9 J.H. Pumell, J.R. Jones and M.H. Wattan, J. Chromatogr., 399 (1987) 99.
10 P. Sandra, F. David, M. Proot, G. Dirricks, M. Verstappe and M. Verzele, J. High Resolut.
Chromatogr. Chromatogr. Commun., 8 (1985) 782.
1 1 G. Schomburg, LC-GC Mag., 4 (1987) 304.
12 U.K. Goekeler and F. Mueller, Process multidimensional gas chromatography, in: Multidi-
mensional Chromatography, Techniques and Applications (H.J. Cortes, Ed.), Marcel Dekker,
New York (1990), p. 191.
13 W. Bertsch, J. High Resolut. Chromatogr. Chromatogr. Commun., 1 (1978) 85.
14 J.C. Giddings, Anal. Chem., 56 (1984) 1258A.
15 G. Schomburg, Multidimensional gas chromatography as sampling technique, in: Sample
Introduction in Capillary Gas Chromatography, Vol. 1, Chromatographic Methods (P. Sandra,
Ed.), Dr. Alfrd Huethig (1985), p. 235.
16 R. Consden, A.H. Gordon and A.J.P. Martin, Biochem. J., 38 (1944) 244.
17 W.D. Snyder, Multidimensional gas chromatography and hyphenated techniques, in: High
Resolution Gas Chromatography (K.J. Myrer, Ed.), Hewlett Packard (1989).
18 W. Engewald and T. Maurer, J. Chromatogr., 520 (1990) 3.
19 A. Zlatkis, J.W. Anderson and G. Holzer, J. Chromatogr., 142 (1977) 127.
20 E. Kugler, W. Halany, R. Schlenkermann, H. Webel and R. Langlais, Chromatographia, 10
(1977) 438.
21 J.F.K. Huber and H.C. Smit, Z. Anal. Chem., 245 (1969) 84.
22 Y.P. Chizhkov and O.A. Yushina, J. Anal. Chem. USSR 31 (1976) 1, Ch. 10.
23 D.R. Deans, Chromatographia., 1 (1968) 18.
24 R. W. Slack and A. C. Heim, Int. Lab., 16 (1986) 88.
25 G. Schomburg, H. Husmann and E. Hubinger, J. High Resolut. Chromatogr. Chromatogr.
Commun., 8 (1985) 395.
26 J.H. Pumell and P.S. Williams, J. Chromatogr., 325 (1985) 1.
27 T.S. Buys and T.W. Smuts, J. High Resolut. Chromatogr. Chromatogr. Commun., 3 (1980)
461.
28 D. Repka, J. Krupcik, E. Benicka, T. Maurer and W. Engewald, J. High Resolut. Chromatogr.
Chromatogr. Commun., 13 (1990) 333.
29 R.E. Kaiser, R.I. Rieder, Lin Leming, L. Blomberg and P. Kusz, J. High Resolut. Chromatogr.
Chromatogr. Commun., 8 (1985) 580.
30 R. Villalobos, R.O. Brace and T. Jahns, in: Gas Chromatography (H.J. Noebbels, R.F. Wall
and N. Brenner, Eds.), Academic Press, New York (1961), p. 39.
31 V.P. Chizhkov, G.A. Yushina, L.A. Sinitzina and B.A. Rudenko, J. Chromatogr., 120 (1976)
35.
32 R.F. Wall, W.J. Baker, T.L. Zinn and J.F. Combs, Ann. N.Y. Acad. Sci., 72 (1959) 739.
33 D.R. Deans, J. Chromatogr., 203 (1981) 19.
34 Deutsche Patentschrift DE-PS 2955-387, 07.12.1976.
35 F. Mueller and M. Oreans, Chromatographia, 10 (1977) 473.
36 K. Grob Jr. and J. Chromatogr., 237 (1982) 15.
37 G. Schomburg, E. Bastian, H. Behlau, H. Husmann, F. Weeke, M. Oreans and F. Mueller, J.
High Resolut. Chromatogr. Chromatogr. Commun., 7 (1984) 4.
38 F. Mueller, H. Mueller and H. Straub, J. Chromatogr., 477 (1989) 25.
39 K. Grob Jr. and H.P. Neukom, J. High Resolut. Chromatogr. Chromatogr. Commun., 2 (1979)
15.
40 K. Grob Jr. and S. Rennhard, J. High Resolut. Chromatogr. Chromatogr. Commun., 3 (1980)
627.
268 Chapter 9
4 1 D.C. Fenimore, R. R. Freeman and P.R. Loy, Anal. Chem., 45 (1973) 233 1.
42 G. Schomburg, H. Husmann and F. Weeke, J. Chromatogr., 99 (1974) 63.
43 M. Oreans, F. Muller, D. Leonhard, A. Heim, Analysis of Volatiles, W. de Gruyter, Berlin
(1984), p. 171.
E.R. Adlard (Ed.), Chromatography in the Petroleum Industty
CHAPTER 10
Supercritical jluid extraction
T.P. Lynch
Anabtical and Applied Science Division, BP Group Research and Engineering Centre, Chertsey Road,
Sunbury-on Thames, Middlesex, UK
10.1 INTRODUCTION
When a substance is maintained at a temperature above its critical point, the

vapour and liquid have the same density and it becomes impossible to liquefy it
by increasing pressure [l]. Thus, a single phase is maintained regardless of the
applied pressure. This single phase is called a supercritical fluid and is character-
ized by a critical temperature (T,) and a critical pressure (PJ. The critical para-
meters for some fluids which have been employed for analytical studies are
listed in Table 10.1.
The supercritical phase has unique properties which can be employed to great
advantage in many processes requiring selective solvation. Basically, a super-
critical fluid has the beneficial properties of both liquids and gases. It has a
density approaching that of the liquid and therefore a high solubilizing power.
The diffusivity is intermediate between a liquid and a gas and the viscosity is
similar to a gas. Furthermore, the density of the fluid, and hence its solvating
power, can be varied by changing the applied pressure. Thus, a supercritical
fluid can be considered as a solvent with continuously adjustable solvent power
and this can be utilized to great advantage in separation processes both on an
industrial and analytical scale. The theory, development and application of su-
percritical fluid technology has beenhe subject of much research in recent years
and several texts [2-6] have appeared which cover these subjects in detail. All of
these texts are recommended to anyone considering working with supercritical
fluids. In this text, we are specifically concerned with analysis in the petroleum
industry and it is interesting to briefly consider the contribution made by this
industry to much of the early research in supercritical fluids both in analytical
and process applications.

270 Chapter 10
ThSLE 10.1
CRITICAL PARAMETERS FOR SOME FLUIDS USED IN ANALYTICAL SUPERCRITICAL
FLUID TECHNIOUES. (DATA TAKEN FROM SF SOLVERTMPACKAGE, ISCO, USA)
Fluid Critical temperature Critical pressure

Tc (C) pc (atm)
Xenon 16.7 57.6

Carbon dioxide 31.1 72.8
Nitrous oxide 36.6 71.5
Sulphur hexafluoride 45.7 37.1
Propane 96.8 41.9
Ammonia 132.4 112.0
Pentane 196.6 33.3
A review of patents relating to supercritical fluids will show a large number

relating to the oil industry. For example, one of the earliest patents was filed in
1927 and referred to a Process for Treating, Separating, and Purifying Oils. In
this patent, the authors refer to a separation where 5 kg of mineral lubricating oil
with an original viscosity of 5.2 cps was treated with 15 kg of liquid carbon di-
oxide. The carbon dioxide extract was found to yield 0.5 kg of oil with a viscos-
ity of 2.2 cps while the residual 4.5 kg had a viscosity of 5.9 cps. This patent is
referred to by many of the subsequent petroleum-based patents including that
assigned to Shell Development Company, San Francisco, CA, in 1940 which is
the earliest found by McHugh and Krukonis [2] which refers to the use of a su-
percritical fluid.
However, perhaps the best known process which relates to the successful ex-
ploitation of the dramatic changes in properties which can occur around the
critical point is the technique of propane deasphalting which is still in use today
for refining oils. The oil industry is also much in evidence when the develop-
ment of analytical supercritical fluid techniques is considered. It is generally
agreed that the first demonstration of supercritical fluid chromatography (SFC)
was by Klesper et al. [7] for the separation of porphyrins, compound types which
are of great interest to the petroleum industry. Furthermore, Sie and Rijnders [S-
113 of Shell Laboratories in Amsterdam are widely credited with much of the
pioneering development of SFC and in fact with using the term supercritical
fluid chromatography for the first time [12]. Following this, Jentoft and Gouw at
Chevron reported the application of pressure programming [13] and mobile
phase modifiers [14]. So, we can see that the oil industry has been a major force
in the development of all aspects of the application of supercritical fluids. This
text, however, is directed at the application of chromatography in the petroleum
industry and as the more mature area of supercritical fluid chromatography
Supercritical fluid extraction 27 1
(SFC) is dealt with in another chapter, the remainder of this chapter is devoted to
the more recent technique of supercritical fluid extraction (SFE) which is cur-
rently being proposed as the sample preparation method of choice for many ap-
plications.
10.2 WHY USE SUPERCRITICAL FLUID EXTRACTION?
Supercritical fluid extraction has become a favoured means of analytical

sample preparation in recent years, particularly for the removal of analytes of
interest from solid matrices prior to chromatography. Although the petroleum
industry deals mainly with liquids, there are many areas where the analysis of
solid matrices is of interest. But why should we choose to employ a supercritical
fluid when traditional methods such as distillation and solvent extraction have
sufficed for many years?
To answer this, we have to again consider the properties of supercritical fluids
and what these mean in terms of the ideal features of an extraction solvent.
10.2.1 Technical advantages
10.2.1.1 Solubility
The first requirement for an extraction solvent is that it must dissolve the
species of interest and the great feature of supercritical fluids is that they have
liquid-like densities and therefore liquid-like solvent powers. The variation of
solubility with density for naphthalene in carbon dioxide at 55C is shown in
Fig. 10.1 and we can see a two order of magnitude increase in solubility on go-
ing from a density of 0.6 to 2.1. One problem that has been encountered in the
use of single component supercritical fluids is that those with easily attainable
critical parameters are relatively non-polar and the type of components which
can be solubilized are therefore also limited by their polarity. This problem can
be partially overcome by using solvent modifiers. These are polar solvents
such as methanol which are added to supercritical fluids such as carbon dioxide
to alter its polarity and hence enhance the solubility of polar compounds in the
supercritical mixture. The mechanism by which these modifiers work is complex
and still the subject of much research but is thought to involve processes such as
hydrogen bonding and molecular clustering [ 151.
10.2.1.2Selectivity
With conventional solvent extraction, the only variable which can be readily
altered to change solubility significantly is temperature. However, with a super-

272 Chapter 10
SF-Sotuer Program
Temp : 68.48 "C Density in g d a t

I
I
- Analytes - 2.5
4t Naae TenpOC
1 Naphtha1 55.88
D
E
N
S
I
T
Y
- Comands
(T ) a h l a t e
=
1 Print iny . . .
(Ehit
Solubi 1i t 9 R .133
Fig. 10.1. Plot of variation of so1ubilit)l of naphthalene with density in carbon dioxide at 55C.
(Reproduced from SF-Solver Software package courtesy ISCO Inc. USA.)
critical fluid, the solvent power can be adjusted over a wide range by varying the
pressure and therefore the density as illustrated in Fig. 10.1. This means that the
solvent power and hence the selectivity of the solvent can be fine tuned to target
the analytes of interest. Table 10.2 lists data where the density of carbon dioxide
has been calculated for various pressures at 35C and 55C. The calculated
density has then been allocated a value for Hildebrand solubility parameter and a
solvent with a comparable value selected. Although this is a somewhat simplistic
approach, it does illustrate that the solvent power of a supercritical fluid can be
easily mmipulated by changing the applied pressure.
Furthermore, the combination of pressure and temperature can also be em-
ployed to obtain selectivity as illustrated in Fig. 10.2 where the variation of
solubility with pressure for naphthalene in carbon dioxide is shown at 35C and
55C. At a pressure of 90 atm, an increase in temperature from 35C to 55C
results in a decrease in solubility and naphthalene would be precipitated from
solution. In contrast, however, at a pressure of 200 atm, the reverse is true and an
Supercritical fluid extraction 273
TABLE 10.2
PRESSURE, DENSITY, HILDEBRAND SOLUBILITY PARAMETER, AND EQUIVALENT
SOLVENT DATA AS PRODUCED BY THE SF-SOLVER COMPUTER PACKAGE, (ISCO,
USA)
~~ ~
Pressure Pressure Density Hildebrand Equivalent

at 35C (bar) at 55C (bar) (g/ml) solubility solvent
parameter
78 100 0.329 2.81 Hydrogen

133 235 0.800 6.82 Methane
155 265 0.828 7.07 n-Pentane
180 300 0.860 7.33 n-Hexane
220 350 0.888 7.57 n-Octane
360 520 0.936 8.21 Cyclohexane
480 >660 1.009 8.60 Carbon tetrachloride
SF-So 1 uer Program Isco , Inc .
Teap 68.48 "C Pressure in ATM

I I
- Analytes -- 284.8
U Hare TerpOC
1 Naphthal 35.88
2 Naphthal 55.88 P
R
E
s
S
0
R
rn Connands
(Tlabulate
Printiny . . .
- E
IElxit a1.1
8.881 Solub i 1 i t y El.133
Fig. 10.2. Plots of variation of solubility of naphthalene with pressure in carbon dioxide at 35C
and 55C. (Reproduced from SF-Solver Software package courtesy ISCO Inc USA.)

274 Chapter I 0
increase in temperature causes a dramatic increase in solubility. This tunability

of solvent power means that one supercritical fluid can be used to extract a wide
range of analytes. For more complete discussions on the factors affecting solu-
bility in supercritical fluids, references [ 2 4 ] are recommended.
10.2.I . 3 Volatility
This may seem an unusual benefit, but in almost all cases the analyst wishes
to perform some further analysis on the extracted species and it is useful to be
able to isolate the extract from the solvent. This can be particularly true for trace
analysis when it is useful to be able to get the analytes in as concentrated a form
as possible in order that they can be measured with sufficient sensitivity by the
chosen analytical technique.
In conventional solvent extraction, this normally means removal of the solvent
by vacuum distillation using a rotary evaporator but this can introduce problems
with the loss of analytes by both evaporation and thermal lability. It can also be a
problem in that trace impurities in the solvent are concentrated along with the
analytes and can interfere with their subsequent determination. In the case of
SFE, the most common solvents are gases at room temperature and below and as
such, on depressurization from the extraction system, are easily removed from
even a cryogenically cooled analyte collection vessel without the need for distil-
lation or the application of heat. Furthermore, as gases they are generally easier
to obtain in a high degree of purity and therefore problems with solvent impuri-
ties can be reduced. This ability to remove the solvent easily is particularly im-
portant when SFE is coupled on-line to chromatographic techniques such as gas,
liquid, or supercritical fluid chromatography.
10.2.I . 4 Mntrix penetration
In the majority of cases where an extraction technique is used, the sample is a
solid and if the chosen solvent has sufficient power to dissolve the analyte(s) of
interest, then the rate determining step in the extraction process will be governed
by how well the solvent interacts with the matrix and removes the analyte from
it. This is where the gas-like physicochemical properties of a supercritical fluid
such as diffusivity, viscosity and surface tension begin to show benefit. The dif-
fusivity of supercritical fluids is typically 1-2 orders of magnitude greater than
that of solutes in liquids and this gives a more rapid transfer of material from a
solid surface to the extract phase compared to that with liquids. In addition, the
viscosity of the supercritical phase is typically an order of magnitude lower than
that of a liquid and this combined with a zero surface tension allows easy pene-
tration of microporous matrices. These factors, when combined, typically give
extraction times which are an order of magnitude less than an equivalent solvent
extraction. The use of temperature can also be influential on the release of ana-
lyte from the sample matrix, particularly when volatile compounds such as petro-
leum-based components are involved. Temperature can also be employed to ef-
fect a structural change in the sample matrix, e.g. melting or converting crystal-
line polymers into amorphous ones, thereby allowing penetration of the matrix
by the solvent. The use of solvent modifiers as mentioned in the section on
solubility can also have an important part to play in matrix interaction where
they can act to displace molecules of analyte which are adsorbed on active sites
on the sample matrix.
10.2.2 Environmental and safety advantages
Carbon dioxide is by far the most popular analytical supercritical fluid and
probably the biggest driving force for the application of analytical supercritical
fluid extraction has been as a result of the benign nature of this fluid. It is cheap,
non-flammable, non-toxic, and is not considered as environmentally unfriendly.
As a result there has been an initiative in the United States by the Environmental
Protection Agency (EPA) to employ it to replace the environmentally unfriendly
and often toxic solvents used in analytical methods which employ solvent ex-
traction procedures. This is becoming more and more important as the evidence
for the toxicity of once common solvents such as chloroform and more recently
hexane has been collected. In addition, the removal of the solvent as gaseous
carbon dioxide means that the costly problems associated with the storage and
disposal of conventional solvents are avoided.
So we can conclude that the main reasons for using supercritical fluid extrac-
tion can be summarized as:
- reduces sample preparation time

- selective extractions are possible
- reduces chemical exposure
- minimizes solvent purchase and disposal costs
- compatible with coupling on-line to chromatography
Let us now look at the equipment and techniques employed to perform SFE
and some petroleum related applications, with particular reference to the use of
SFE as a pre-separation technique for chromatography.
10.3 SFE EQUIPMENT
The basic requirements for equipment for analytical scale SFE are as follows:

216 Chapter 10
--
FEED FLUID
~I-
Fig. 10.3. Schematic representation of a simple SFE system.
- a supply of fluid at the required pressure

- a pressure-safe sample vessel
- a means of controlling the extraction temperature
- a depressurization device
- a means of collecting the extract on depressurization
The coupling of these requirements is shown schematically in Fig. 10.3. In or-

der to fully describe all the variants of each component which are available and
their relative merits and disadvantages would require a chapter on its own, so we
will only briefly cover the basics here. Readers who wish to find out more are
advised to consult the specialist texts recommended earlier or to contact all the
manufacturers and decide for themselves which product best suits their require-
ments.
10.3.1 Pumping systems
The pumping system is the heart of an SFE system. It must be able to supply a
continuous stream of fluid at the desired pressure and preferably at a user-
selectable flow rate. There are two main types of pumps in use at present,
namely syringe and reciprocating piston pumps. Syringe pumps were primarily
developed for SFC where the main requirement is for a pump which can be pres-
sure programmed but also provide a pulseless controlled flow of fluid. However,
they do have limitations for use in SFE, namely they are relatively expensive,
and have a limited volume (typically between 150 and 300 ml) and unless two
are used in tandem, the extraction may have to be stopped to refill the pump. In
addition, they are difficult to employ with solvent modifiers.
Reciprocating piston pumps of the type commonly used in HPLC can easily
be modified to pump supercritical fluids such as carbon dioxide [ 16,171. They
can be easily coupled together to provide binary or even ternary solvent systems
as in HPLC, and will provide a continuous flow of fluid at high flow rates. They
do not give a pulseless flow like a syringe pump but this is not as important in
SFE as it is in both HPLC and SFC. One drawback of employing HPLC pumps
to pump carbon dioxide is that it is usually necessary to chill the pump head and
the feed carbon dioxide to ensure that it remains as a liquid. However, recently a
new syringe type pump has appeared [ 181 which is specifically designed for use
with analytical SFE and this does not require the pump head to be chilled. An-
other type of piston pump which is air driven has also found uses where flam-
mable or corrosive solvents have been used. This type of pump has a high ca-
pacity condenser on the inlet which can liquefy gases such as propane, ethylene,
and ammonia [ 191.
10.3.2 Sample vessels and temperature control
The main requirement for sample vessels is that they must be able to safely
withstand the pressures and temperatures used in extraction. In addition, they
should be easy to open, load and seal, and to connect into the extraction system.
Ideally they should have a low thermal mass so that they reach thermal equilib-
rium quickly. Early cells were empty HPLC columns but these were normally
small diameter and it was difficult to fill and remove the sample. More recently,
custom designed cells with removable end frits and finger tight connections have
been developed which are available in sizes ranging from a few hundred microli-
tres to about 50 ml. It is also necessary to be able to maintain the temperature of
the sample cell at the required value and this is most commonly done using a
fan-assisted oven to facilitate rapid heating and cooling of the extraction cell. In
the early days of SFE, one of the main claimed advantages of the technique was
that it worked best at low temperatures (i.e. high density) and was therefore
considered useful for thermally labile substances. As a result, many of the ovens
used had a maximum temperature of 100C. However, recent work has shown
that temperatures in the region of 200C are often needed to obtain maximum
analyte recovery of PAHs and this should be considered when looking at sys-
tems. In addition, if chilled pump heads are being used, the temperature of the
fluid may be low and it is worth having some mechanism to pre-heat the fluid to
the desired temperature before it enters the extraction vessel.

278 Chapter 10
10.3.3 Depressurization
There are a number of devices which allow the depressurization of the solvent
from the extraction pressure to atmospheric pressure and therefore cause precipi-
tation of the extract in the collection device. Until recently, the most popular
device was one which provided a restriction to the flow of the fluid by passing it
through a narrow tube or orifice, therefore building a back pressure. The most
common way of achieving this used a fused silica capillary to restrict the escape
of the gas and therefore maintained pressure in the extraction cell. The pressure
which was maintained was dependent on the length and internal diameter of the
restrictor. The main problem with these restrictors was that the flow could not be
controlled independently of pressure. In addition, they were fragile and prone to
blockage with extract. Other variants employed crimped metal tubing and frits in
the end of tubing but they suffered from the same problems.
It is amazing to think that a technique which relies on pressure and pressure
control for its success employed such primitive technology. The second type of
device which has been employed is the back pressure regulator. These were
originally based on pressure relief type valves where the force exerted by a
spring was used to force a needle into a seat. When the system pressure is suffi-
cient to lift the valve it was released. The pressure could be set by adjusting the
spring force via a screw thread. These systems gave independent control of flow
and pressure but were still prone to blockage and sticking. Saito et al. [20] made
a significant breakthrough by designing an electronically controlled high speed
switching valve. The configuration of the valve was such that precipitated ex-
tract and solid carbon dioxide was cleared from the valve by the vibration and
punching action of the valve needle. This and other electronically controlled
valves which have been developed since have brought analytical SFE to the
stage where analysts in different laboratories can repeat exactly the same ex-
periment in terms of pressure, temperature and flow rate and expect to get the
same result.
10.3.4 Extract collection
In the early days of analytical SFE development this was one of the most ne-
glected areas. Indeed, in much of the early work on SFE which reported low ex-
traction efficiencies, it was the trapping on depressurization which was the
problem and not incomplete extraction. This is not surprising when one consid-
ers that the flow of solvent gas on depressurization is often in the litre/minute
range and that the precipitated extract is in the form of a fine aerosol in the gas
phase. Since these early days, many techniques have been developed to ensure
complete collection of the extract and some of the available options are listed
below:
- in an empty glass vial

- onto a solid surface
- on an inert solid, e.g. glass balls
- on an active solid sorbent, e.g. C18 stationary phase
- bubbling into solvent
The efficiency of all of these methods can often be improved when they are
combined with each other, e.g. glass balls in a solvent to maximize surface area,
or combined with cryogenic cooling of the trapping material. There have been
many studies made on the efficiencies of the different trapping methods [21-251
and there are a number of factors to consider when selecting the best for a spe-
cific application. The chemical nature of the analyte is important as is the ana-
lytical method which will be used to analyse the extract. One disadvantage of the
solid-based traps is that most subsequent analytical methods (e.g. GC or HPLC)
require a solution and therefore it is necessary to then carry out a solvent extrac-
tion, albeit on a smaller scale, to get the analytes from the trap into solution.
Also if a solvent modifier is used, it may be impossible to use solid type traps as
the precipitated co-solvent washes the analyte from the trap and losses can oc-
cur. In such cases, it is probably best to use the modifier solvent as the trapping
solvent.
One way of studying trapping efficiency is the use of spiked samples. In
these, known amounts of pure components, usually in solution, are added either
to an inert support or to a clean sample of the matrix of interest. These Sam-
ples are then analysed using the proposed method and the recoveries of the
spiked analytes calculated. These type of results are often shown in publications
as a proof of complete extraction. This is not valid as spiked samples cannot
mimic the way in which real analytes interact with the matrix, particularly when
the samples have been aged and weathered as in many environmental samples
such as soil. However, they can be used to check the integrity of the extraction
and trapping systems and as such are very useful tools. In fact it is recommended
that this is done on a regular basis as a quality control check for routine analyti-
cal SFE methods.
One other approach, which is particularly useful for samples which will be
analysed by chromatography, is to employ internal standards which are added to
the sample in the cell and then used both as a means of quantification of the
analyte and as a means of checking the efficiency of the system plumbing and
trapping system. For example, in looking at the concentration of low molecular
weight oligomers in the range C8 to C20 in polyethylene, it is possible to employ

280 Chapter 10
an internal standard which contains three normal hydrocarbons as internal stan-

dards. This is possible because the oligomers are all even carbon numbers and
therefore all the odd carbon numbers are available for use as internal standards.
So a solution of known concentration of three normal hydrocarbons of odd car-
bon number which cover the range of interest, e.g. nC9, nC13, and nC19 is
added to the sample in the extraction cell and the extraction carried out. The re-
sulting extract is analysed and the response of the intermediate standard checked
against a direct analysis of the internal standard solution. This standard is then
used to calculate the concentration of the other components in the extract. The
efficiency of analyte trapping can then be assessed over the whole range of in-
terest by checking the recoveries of the other two internal standards.
10.4 SFE TECHNIQUES
We now consider some of the techniques employed to actually carry out the
extractions.
10.4.1 Static extraction
In static extraction, the sample cell is pressurized with fluid and the sample is
allowed to steep in the solvent until equilibrium is reached. An aliquot of the
extract is then removed, or the system is then depressurized and the analyte re-
covered. The main benefit of this method is that the fluid has time to penetrate
the matrix . It is most applicable when the analyte has a high affinity for the sol-
vent and a low affinity for the matrix and also when the solubility limit of the
analyte in the fluid is much higher than the actual level reached during the ex-
traction.
10.4.2 Dynamic extraction
This is the most common mode of operation for analytical SFE and a typical
flow manifold for operation in this way is shown in Fig. 10.3. New fluid is con-
tinuously flushed through the sample and out through the back pressure device
into the trap. This is a particularly useful mode when the concentration of the
solute in the sample is high relative to its solubility in the fluid or when the sol-
ute has a high affinity for the sample matrix. Many modem instruments can op-
erate in both dynamic and static modes and often the best approach is to combine
both in the same method with a static step followed by a dynamic step.
Supercritical fluid extraction 28 1
OVEN EXTRACTION
CELL
3
YV
FEED L
SAMPLING
VALVE -
PUMP -' EXTRACT
J
Ak 1r
FEED FLUID EXTRACTION

RECIRCULATING,-
MONITOR
PUMP
Fig. 10.4. Schematic representation of a recirculating extraction manifold.
10.4.3 Recirculating extraction
This combines some of the features of both static and dynamic extraction. In
this mode, the sample cell is part of a loop which is pressurized with the super-
critical fluid. The loop also contains a recirculating pump which can pump the
fluid around the loop and over the sample as in dynamic extraction. A flow dia-
gram of a typical recirculating system is shown in Fig. 10.4.
Alternatively the sample can be allowed to steep in the fluid as in static ex-
traction. These steps are often alternated several times until the system reaches
equilibrium. The extract can then be collected by depressurization of the loop or
an aliquot of known volume can be removed via a sampling valve for further
analysis. This can be a useful means of studying the effect of temperature and
pressure on solubility as only a small volume is removed in relation to the total
loop volume. Therefore experiments can be carried out and the solubility calcu-
lated at several different temperatures and pressures on a single sample. This
method, however, is not generally available on commercial instruments and has
not found many published analytical applications.
10.4.4 Extraction of liquids
Conventional SFE extraction cells are often not suited for the direct extraction

282 Chapter 10
of liquids as the supercritical fluid can tend to pump the liquid through the sys-
tem rather than extract it. This is further complicated by the fact that in many
cases, two solutions are present during the extraction process; one is a solution
of extract in the fluid and the other a solution of the fluid in the matrix. The sec-
ond effect can cause a significant increase in volume of the sample, particularly
on depressurization as dissolved gas is liberated, and if the cell is too full, the
contents can be carried through to the extract collection system. This is unfortu-
nate for the extraction of oil samples but several methods have been devised for
the extraction of such samples.
These include:
- modified extraction cells

- immobilization on a solid sorbent
- inserts for conventional cells
Taylor et al. have modified conventional extraction cells and described their
use for the extraction of liquids including the extraction of phenols [26] and
phosphonates [27] from water. The use of solid adsorbents has been employed
by many workers for the extraction of liquid samples. Here the liquid sample is
placed on a solid substrate to immobilize it. This method has the advantage that
it can be used to impart a greater degree of selectivity to the extraction process
as the sorbent can be chosen such that it selectively interacts with components in
the sample. A typical example is the analysis of PCBs in electrical oils [28]
where the oil is adsorbed onto Florisil for the extraction process. A method
which we have successfully employed in our laboratories for the extraction of
END SECTION
LO S G IT L D I XA L S E C T I0 N
Fig. 10.5. Schematic of the boat in cell method for extracting liquids. (Reproduced courtesy of
British Petroleum.)
Supercritical jluid extraction 283
oils employs glass boats which are inserted into a conventional extraction cell. A
schematic representation of the system is shown in Fig. 10.5. The boats are con-
structed to sit in the cell and the cell is maintained in a horizontal position
throughout the extraction. This type of arrangement has the disadvantage that
only the surface of the oil sample is exposed to the extraction fluid and as a re-
sult extraction times are generally longer. However, the method does have the
advantage when the residue after extraction is the component of interest as it is
contained in the sample boat and can be readily removed for further analysis.
This also minimizes contamination of the extraction vessel and can significantly
reduce the cleaning time required after extraction.
10.4.5 Reactive extraction
This is a relatively new concept where a reagent is added to the sample prior
to extraction. The reagent reacts with the analyte under supercritical conditions
to form a species which is readily extracted by the supercritical fluid. The rea-
gent can also aid in releasing the analyte from the matrix by interacting with ac-
tive sites on the surface. This type of approach began with the application of
common derivatization reactions used in GC and HPLC to improve the solubility
of polar solutes in non-polar carbon dioxide. In most methods, the reagents are
added directly to the sample in the extraction cell and the derivatization is car-
ried out under supercritical conditions. This allows derivatization and extraction
to be performed in a single operation. Hawthorne et al. [29-3 11 reported proce-
dures which produced and extracted methyl esters of components of interest.
Hills et al. [32,33] performed simultaneous silylation and extraction on a sample
of coffee which had already been exhaustively extracted with supercritical car-
bon dioxide and reported the extraction of many new components both silylated
and unsilylated from the matrix. This area of SFE will undoubtedly develop fur-
ther and one could envisage the development of selective extraction agents for
particular analytes and matrices which prove intractable by other techniques.
One area where this would be particularly applicable would be the development
of complexing agents for metals which would give SF-soluble metal complexes
which could then be extracted and determined.
10.4.6 Off-line extraction
Off-line extraction is the term given to methods where the extracted analytes
are collected and isolated in some way not directly coupled to the subsequent
analytical techniques which are to be employed. For example, they can be col-

284 Chapter I0
lected in a solvent or on a solid sorbent. This is the simplest way to perform SFE
and requires much less skill on the part of the analyst. It is therefore the most
common mode of SFE and should be quite adequate for most routine procedures.
In many cases, the most crucial decision in off-line SFE is the choice of the col-
lection methods described earlier.
10.4.7 On-line extraction
On-line extraction is the term given to methods where the extraction process
is coupled directly to the analytical technique which is to be used for the further
analysis of the extract. This is also sometimes called a hyphenated technique
and common examples include SFE-GC, SFE-SFC, SFE-HPLC and SFE-IR.
These methods are generally more difficult to perform than off-line extractions
as it i s vital to get the interface between the two techniques right in order to ob-
tain meaningful results. However, they do have several advantages including the
total transfer of the extract which results in maximum sensitivity. Sample han-
dling and exposure to the atmosphere can be eliminated between extraction and
analysis and this reduces the chances of analyte contamination and degradation.
Furthermore, when employed with combined chromatographic-spectroscopic
techniques such as GC-MS, complex multi-component data can be obtained from
a single experiment.
10.5 PETROLEUM-BASED APPLICATIONS
The prime subject of this book is chromatography and therefore we shall con-
centrate on applications where SFE is employed as a pre-separation technique
for the various forms of chromatography and divide them into the categories of
off-line and on-line applications. The particular applications have been chosen to
cover exploration and production of the crude oil, through processing in the re-
finery, to its final use in automotive engines.
10.5.1 Off-line applications
IO. 5. I . I Residual oil on drill cuttings

The use of oil-based drilling mud offers significant financial advantages over
water-based mud but there are disadvantages in that the drill cuttings produced
become contaminated with the oil from the mud. The levels of oil can be as high
as 10-1 5 weight % and as they can be produced at several tons per hour, the dis-
SupercriticalJuid extraction 285
posal of these cuttings (for example dumped in the sea in an offshore operation)
could have a significant environmental impact. As a result, impending legislation
has fostered several initiatives to try to reduce the oil level of cuttings prior to
disposal. These have included roasting, washing with solvents and super-
wetters, and even by extraction with supercritical fluids [34]. In order to assess
the efficiency of a method of cleaning oil from cuttings, it is necessary to be able
to measure the residual oil. The conventional method employs a retort method
where a weighed sample of cuttings are heated in a vessel and the desorbed oil
and water condensed and collected in a measuring cylinder. The resulting vol-
umes can be read and approximate levels determined. This provides acceptable
accuracy and precision for levels of 10-15% but not at the proposed levels of
1%. This type of application is ideally suited to off-line SFE with carbon dioxide
as the oils used (which are in the diesel oil range) are extremely soluble.
During trials of a clean up system, our laboratory was required to determine
the levels of oil in both the feed and cleaned cuttings at levels down to 0.1%. A
method was developed where the cuttings were extracted with SF carbon dioxide
and the extracted oil quantified by gas chromatography. The schematic represen-
tation of the process is shown in Fig. 10.6. A weighed sample of cuttings is
placed into the extraction vessel and the oil extracted with carbon dioxide using
a dynamic extraction at 350 bar and 80C at a flow rate of 2 ml/min. On depres-
surization via a back pressure regulator, the expanded gas and extracted oil is
passed through a silica Sep-pak cartridge which collects the oil. The Joule
Thomson cooling of the expanding gas acts as a cryogenic coolant for the Sep-
pak cartridge and helps ensure efficient trapping. After the extraction is com-
r
SAMPLE CELL
PRESSURE
REGULATOR
PUMP OVEN
CARBON DIOXIDE FEED U
Fig. 10.6. Schematic manifold for the extraction of drill cuttings to determine residual oil.
(Reproduced courtesy of British Petroleum.)

286 Chapter 10
TABLE 10.3
COMPARISON OF RESULTS OBTAINED BY SFE AND SOXHLET EXTRACTION FOR THE
DETERMINATION OF RESIDUAL OIL ON DRILL CUTTINGS (ALL RESULTS ARE %
WEIGHTNEIGHT)
Sample SFE Soxhlet
A 2.06 1.81
B I .89 1.78
C 0.56 0.62
D 0.58 0.58
plete (approx. 30 min) the Sep-pak is removed and the oil eluted from it using a
few millilitres of carbon disulphide. A known weight of internal standard is then
added and the sample analysed by gas chromatography to quantify the oil. The
method was compared with an 8-h Soxhlet extraction on a number of samples
and the results are shown in Table 10.3.
These results show that the SFE method was comparable to the Soxhlet
method but it did not give any indication on the efficiency of extraction. This
was evaluated by submitting small samples (approx. 2 mg) of powdered drill
chips before and after extraction to thermal desorption gas chromatography
where they were rapidly heated such that all oil was desorbed into the GC. The
results obtained indicated that less than 2% of the oil originally present on the
cuttings was left after the SF extraction, i.e. greater than 98% extraction effi-
ciency was achieved. This method adequately shows the advantages of SFE. A
result could be obtained in less than an hour, nearly an order of magnitude
quicker than Soxhlet extraction. This can be important if the results are being
used to optimize or control a real process as performance variations can be de-
tected earlier and the necessary action taken. Each SFE sample required a few
millilitres of carbon disulphide, whereas each Soxhlet extraction produced a few
hundred millilitres of solvent which had to be stored and then disposed of. In
addition, the accuracy and precision of the method was more than adequate for
the customer requirements.
10.5.I . 2 Drilling mud characterization
SFE with carbon dioxide has been applied to the characterization of hydrocar-
bons in drilling mud [35]. The method involves placing 0.5 g of sample in an
extraction cell with Hydromatrix. The samples were 6 0 4 5 % water and the
Hydromatrix is a drying agent which absorbs the water and prevents it interfer-
ing with the extraction. The sample is extracted with carbon dioxide at 400 atm
and 100C with a compressed fluid flow rate of 2 ml/min. The extract is col-
lected in dichloromethane at -3OOC and then analysed by gas chromatography. A
inject
I
\O
I
20
c14
\
Retention Time (minutes)

J
I
30
I
k
Fig. 10.7. Chromatogram illustrating a high resolution fingerprint of hydrocarbons obtained from
the extraction of a drilling mud. (Courtesy Suprex Corporation.)
chromatogram showing a typical GC fingerprint obtained from a mud is shown

in Fig. 10.7. The SFE method took 30 min to carry out and replaced a Soxhlet
extraction with methylene chloride which typically took 6-8 h.
10.5.1.3 Petroleum core and rock extractions
Both off-line and on-line extraction techniques have been applied to the
analysis of petroleum-rich core and rock samples. Off-line methods have the ad-
vantage that they allow the use of modified solvents which cannot be used in
most on-line methods. For example a 50/50 mix of dichloromethane and metha-
nol has been employed as a modifier (1 5%) in carbon dioxide for the extraction
of oil from cores [36].
Methanol (15%) in carbon dioxide was also employed for the extraction of a
bitumen-rich core sample. Both extractions were performed at a pressure of
TABLE 10.4
COMPARISON OF SFE AND DEAN AND STARK FOR THE EXTRACTION OF
PETROLEUM CORE SAMPLES
The RSDs are determined upon triplicate extractions. (Data reproduced courtesy of Dionex
Corporation)
Sample Dean and Stark wt% SFE wt% SFE % RSD
Ground core 8-9 8.2 2.0

(700 feet deep)
Ground core 10-1 1 11.8 1.7

288 Chapter I0
600 atm and a temperature of 100C. The collection solvent was toluene and the
extraction time was 60 min. The results obtained are given in Table 10.4. Again
superior results were obtained in a shorter time than conventional methods.
10.5.I . 4 Refinery catalvsts, deposits and sludges
Refineries can produce a number of samples which are amenable to analysis
5y SFE including catalysts, deposits and sludges. The examination of micro-
porous catalysts can be important both in the R&D and operational environment
and SFE can be employed to extract compounds which are causing fouling or
poisoning of the pore and active site of the catalyst. Subramaniam et al. [37,38]
employed both reactions and extractions under supercritical conditions to study
the mechanisms of coke formation on a commercial isomerization catalyst. The
formation of deposits in pipework and vessels is also a problem in refinery op-
erations and SFE can be employed in the characterization of such deposits both
in terms of the extractable components and also as a means of removing hydro-
carbons to allow examination of the solid by other techniques such as thermal
analysis and spectroscopic methods where fluorescence from the hydrocarbons
interferes with the analysis. Refinery sludges are monitored for components such
as polycyclic aromatic hydrocarbons to meet environmental concerns and
Schmidt et al. 1391 have employed SFE with carbon dioxide to replace a tedious
solvent extraction procedure for this process. The extraction is performed on
-400 mg of sample in a 1.5-ml vessel using a pressure of 350 bar, a temperature
of 40C,and a flow rate of 3.5 ml/min. A static extraction of 1 min is followed
by a dynamic extraction for 4.2 min. The extract is collected on depressurization
in a trap packed with 3 0 p m ODS column packing which is maintained at 10C
during the extraction. The extract was recovered from the trap by washing with
hexane (2 ml) and then analysed by GC-MS with single ion monitoring to detect
the compounds of interest.
10.5.1.5Automotive engine particidates
Automotive engine particulates include exhaust particulates, particulates
which have accumulated on filters in the engine, and particulates which have
agglomerated and formed deposits on engine components such as valves. They
are often only available in milligram quantities and are therefore difficult to
handle in solvent extraction procedures. However, they are well suited to SF
methods. The analysis of these particulates is important for several reasons in-
cluding the development of fuels and lubricants, and for environmental monitor-
ing.
Bartle et al. [40] have studied the use of SFE for the extraction of polycyclic
aromatic hydrocarbons from diesel exhaust particulates. It has also been applied
to the sequential extraction of fuel and lubricant components from engine filters
[41] in an attempt to relate the results to the cause of engine failure. The extracts
were then characterized by GC. In this case one extraction yielded a 134% re-
covery in comparison to Soxhlet extraction. The extraction with carbon dioxide
was carried out at 329 bar and 60C (a density of 0.85 g/ml). Static extraction for
10 min was followed by dynamic extraction for 11 min at a flow rate of
3 ml/min. The extract was collected on a solid trap containing ODS and main-
tained at 20C. The extract was then recovered for GC analysis by rinsing the
trap with two 1.2-ml volumes of carbon disulphide.
The examination of engine deposits is another area where SFE can be em-
ployed particularly for the comparison of fuels and lubricants and their perform-
ance in minimising deposit formation. The yields obtained from the extraction of
deposits from two identical engines but using different lubricants are shown in
Table 10.5. The deposits were sampled from the piston crown, the cylinder head
and the inlet valve tulips. Sequential extractions were carried out on each sam-
ple. The first was with carbon dioxide only and the second was with carbon di-
oxide containing 10% methanol as solvent modifier. The deposits were exam-
ined before and after extraction by elemental analysis and thermo-gravimetry to
give values for fixed and volatile carbon, hydrogen, oxygen, nitrogen and ash.
Solid state I3C NMR was also employed to give speciation information on the
carbon. The extracts were examined by GC-MS for the volatile components and
thin-layer chromatography (TLC) for the non-volatiles. GC-MS of the carbon
dioxide extract gave a typical lubricating base oil profiles with some substituted
and polycyclic aromatics. The methanol modified extracts contained mainly high
molecular weight aromatics up to six rings and the piston and head deposits
contained oxidized aromatics. TLC of the carbon dioxide extract showed mainly
non-polar material typical of a degraded oil and the valve deposit extract con-
tained antioxidant. The methanol-modified extract covered a wide polarity range
including mineral oil degradation products and additives. All the modified ex-
tracts contained sulphonates and other highly polar components.
TABLE 10.5
EXTRACTION YIELDS IN WEIGHT Yo OBTAINED BY THE SF EXTRACTION OF ENGINE
DEPOSITS FROM PISTON CROWNS, CYLINDER HEADS, AND VALVE TULIPS FROM
TWO IDENTICAL ENGINES USING TWO DIFFERENT OIL FORMULATIONS X AND Y
~~
Extraction Oil X Oil Y

fluid
Piston Head Valve Piston Head Valve
co2 2 4 7 1 1 3
C02h4eOH 8 6 10 6 6 9
Total 10 10 17 7 7 12

290 Chapter 10
Thus, SFE was employed as a tool to help give detailed compositional analy-
sis of small amounts of sample and allowed studies to be carried out to try to
relate deposit formation to fuel and/or lubricant formulation. However, the SFE
results alone can give interesting information in that the deposits from oil X
contain significantly higher carbon dioxide extractables than those with oil Y.
The carbon dioxide extracts contain mainly low polarity base oil and the results
indicate that during this test there may have been a leaky valve seal allowing
more oil to ingress to the valve and therefore the combustion area. A study of the
engine test results confirmed that this was the case. The method could therefore
be a useful tool in investigating the causes of engine failure and validating com-
pensation claims citing oil products as the cause of the failure. In a wider con-
text, the results demonstrate the power of modified solvents in that the addition
of methanol at a level of 10% to carbon dioxide gave up to a sixfold increase in
the quantity of material extracted. It also shows how they can be employed to
gain fractionation of materials by polarity.
10.5.I . 6 Environmental analysis
This is likely to be one of the biggest application areas for SFE, particu-
larly for the analysis of soil samples and there are a great many papers in the lit-
erature which deal with this topic. Therefore it is considered only briefly here
and readers wishing more detailed information are advised to consult the litera-
ture.
In terms of petroleum industry applications, the two main areas are in the de-
termination of total petroleum hydrocarbons (TPH) and polycyclic aromatic hy-
drocarbons (PAH). Lopez-Aviia et al. [42] have developed a method which em-
ploys SFE off-line followed by infrared (IR) spectroscopy for the determination
of petroleum hydrocarbons in soil. The most favoured method at present is sol-
vent extraction with Freon-1 13 and they estimate that at present in the United
States alone, some 200 000 samples per annum are anaiysed in this way requir-
ing 30 000 1 of Freon-1 13. A similar approach using SFE with GC instead of
SFE-IR can also be employed [43]. Campbell and Richards [44] compared SFE
with Soxhlet and sonication methods for the determination of priority pollutants
in soil and concluded that SFE was, in the majority of cases, the most efficient
extraction method. In addition, significant reductions in organic solvent con-
sumption were achieved.
Hawthorne and co-workers have made extensive studies on the extraction of
PAHs from a variety of matrices, including soils, and using different supercriti-
cal fluids. They reported that chlorodifluoromethane yielded consistently higher
extraction efficiencies than either carbon dioxide or nitrous oxide and was par-
ticularly effective in removing water from wet matrix samples. However, it re-
mains to be seen whether such solvents would be universally accepted.
Supercritical jluid extraction 29 1
10.5.2 On-line applications
The advantages of coupling SFE on-line with chromatographic techniques has

stimulated a lot of work in this area. Much of the early work centred on coupling
SFE with SFC and this has been extensively studies particularly for the determi-
nation of low molecular weight components and additives in polymers [45-47].
However, the limited applicability of SFC soon led researchers to investigate
coupling to GC and LC.
10.5.2.1 On-line SFE-GC
Gas chromatography is probably the most widely applied analytical tool in the
petroleum industry and therefore on-line coupled SFE-GC is likely to be of in-
terest to the petroleum chemist. In this section, we briefly look at the history of
the technique and consider one way of coupling SFE to GC. A more detailed
discussion on the various approaches to coupling SFE and GC is given by Lee
and Markides [4]. Smith et al. [48] described a means of coupling SFE to GC
and described its use for the examination of polycyclic aromatics in a number of
matrices. Monin et al. [49] described the application of SFE-GC for the extrac-
tion and examination of hydrocarbons from source rocks. Hawthorne et al. [50-
531 and Levy et al. [54-561 have published extensively on the different mecha-
nisms of coupling SFE with GC and have demonstrated the application of the
technique for a number of petroleum-related applications.
c02
I GAS CHROMATOGR H
Fig. 10.8. Schematic representation of coupled SFE-GC with direct expansion into a split injector
port.

292 Chapter 10
Supercritical Fluld (1.e.CO,)

I
Extraction Vessel
GC Split Injector
Split Vent
Capillary GC Column
Fig. 10.9. Schematic representation of an on-line SFE-GC interface with a split injector. (Courtesy
of' Suprex Corporation.)
The most generally applicable method of coupling the SFE and GC has been
by inserting the restrictor from the SFE system directly into the injector of the
gas chromatograph. Both on-column and split-splitless injectors have been suc-
cessfully employed with the latter being the most flexible and widely applicable.
A schematic representation of an SFE-GC system is shown in Fig. 10.8 and a
more detailed schematic of the interface using a split injector is shown in Fig.
10.9. In this approach, the supercritical state is maintained until it reaches the tip
of the transfer line and then decompresses directly inside the heated injection
port. The heat of the injection port aids in minimizing the Joule Thomson ex-
pansive cooling effect and also vaporizes the analytes which are then carried
Supercriticalfluid extraction 293
% Peak area
16 - vI
14
12
10
8
6
4
2
0
- nC5 nC6
Direct inlection
015 min extraction

nC7 nC8
Alkanes
!ES
nC9 nClO
6 min extraction
3 0 min extraction
nC11 nC12
Fig. 10.10. Recoveries of nC5 to nC12 standards by coupled SFE-GC. The extraction was inter-
faced to the chromatograph via expansion of the extract into the split injector. The GC oven was
maintained at 4 0 C by cryogenic cooling. Results for direct injection without SFE are presented
together with results for three different extraction times for each component. (Reproduced courtesy
of British Petroleum.)
onto the GC column where they are trapped by the GC stationary phase. The GC
oven is often cooled cryogenically to aid the focusing of the analytes on the sta-
tionary phase. The capabilities of such a system is illustrated in Fig. 10.10 where
the recoveries for a mixture of nC5 to nC12 hydrocarbons are shown. The stan-
dard mixture was spiked on sand and then extracted with carbon dioxide. The
extract was expanded directly inside a split injection port and trapped on the GC
capillary column by cryogenically cooling the GC oven to 4 0 C . The GC was
then temperature programmed and the area of the eluting peaks measured. The
normalized area percent for each peak is represented by a bar and data resulting
from experiments carried out at three different extraction times is compared with
the results obtained by direct injection of the standard solution without SFE. The
results show that in each case, the recoveries were very good with the lowest
being nC5 at -80%. The results were largely independent of extraction time and
this indicates that the method could be applicable for the analysis of real sample
matrices where the time required to achieve quantitative extraction is generally
longer due to the effects of the sample matrix.
In many cases it is difficult to identify a11 the components which are extracted
from samples by the flame ionization detector response alone. This is a particu-
lar problem when the sample history, and/or the identity of the compounds of
interest, are not known, e.g. when examining a catalyst to try and identify poi-

294 Chapter I0
sons. It is in these cases that multi-hyphenated techniques such as SFE-GC-MS,

and SFE-GC-FTIR-MS can be particularly powerful tools.
SFE-GC with spectroscopic detection for the identification of pollutants in soil.

The examination of soil samples for pollution is important in many sectors of the
oil industry. For example, it is important to ensure that land which has, or has
had, oil installations sited on it is free from toxic chemicals such as PAHs to
minimize the risk to workers during excavations. It is also useful during clean up
operations to be able to determine the extent of the contamination.
Conventional methods for the extraction and analysis of such samples are
time-consuming and expensive but SFE-GC combined with spectroscopic detec-
100 1
90 -
h
80 -
8 70-
v
-
a, 6 0 -
2 50-
* 40-
-
u. 30-
7
20 -
0
lo j I I I I I I I I I
600 800 1000 1200 1400 1600 1800 2000 2200
Scan number
100 b 1573 I 1603

1 ;
80 -
I
v
70-
-
a, 60-
50-
-
cn 40-
30-
20 -
10 -
0-
I I I I I I I
1200 1300 1400 1500 1600 1700 1800
Scan number
Fig. 10.1 I . (a) Total ion chromatogram obtained from SFE-GC-MS of a soil sample showing the
presence of polycyclic aromatic hydrocarbons (PAHs). (b) Expanded portion of chromatogram (a)
with library spectral matches for PAHs. (Reproduced courtesy of British Petroleum.)
Supercritical jluid extraction 295
t h
128
Naphthalene
I I I I I I I I
800 900 1000 1100 1200 1300 1400 1500
Scan number
t 5 ring
eg benzo pyrenes
b
.-E
v) I 4 ring
eg chrysenes
A
I
228
9 4 ring
eg pyrenes,
fluoranthenes
202
t
3 ring 17%
eg anthracenes,
phenanthrenes
, h A A , , ,.._.
I I I I I I I I
800 900 1000 1100 1200 1300 1400 1500

Scan number
Fig. 10.12. (a) Chromatograms obtained by single ion monitoring for substituted naphthalenes in a
soil sample by on-line SFE-GC-MS. (b) Chromatograms obtained by single ion monitoring for 3-,
4- and 5-ring PAHs in a soil sample by on-line SFE-GC-MS. (Reproduced courtesy of British Pe-
troleum.)
tion can be a powerful tool for screening samples to decide whether they require
the full analysis. For example, Fig. 10.1l(a) shows the total ion chromatogram
obtained by SFE-GC-MS of a soil sample from a petroleum installation. Exami-
nation of the spectra for each peak has identified the presence of polycyclic aro-
matic hydrocarbons as shown in the expanded portion of the chromatogram in
Fig. lO.ll(b). The technique can be further refined by performing single ion
scans for ions which are characteristic of particular PAHs. Examples from the
examination of a soil sample are shown in Fig. 10.12(a) where ions typical of
substituted naphthalenes are monitored and in Fig. 10.12(b) where ions charac-
teristic of 3-5-ring PAHs are monitored.

296 Chapter I 0
TABLE 10.6
SINGLE ION COUNTS FOR NAPHTHALENE (PARENT) AND C1, C2 AND C3
SUBSTITUTED NAPHTHALENES OBTAINED BY ON-LINE SFE-GC-MS FOR FOUR SOIL
SAMPLES TAKEN FROM THE SAME SITE
~
Sample Parent ion Cl c2 c3

no. (M/W 128) (MIW 142) (MIW 156) (M/W 170)
i 1000 109 66 0
2 787 73 0 0
3 120 0 0 0
4 0 0 0 0
Although this has not yet been shown to be fully quantitative, it can be em-
ployed as a rapid screening process to determine the extent of contamination and
to give a guide as to whether samples should be submitted for more detailed
quantitative analysis. The results listed in Tables 10.6 and 10.7 show the ion
counts for single ion monitoring for selected PAHs for a number of soil samples
taken from the same site. These results show that the technique is at least semi-
quantitative and can be employed to determine the extent of contamination on a
site, or in cases of dispute, identify the source of the contamination. Each analy-
sis can be run in under 2 h although data analysis may take longer if a lot of
component spectra have to be examined. Figure 10.13(a) shows the total ion
chromatogram from another soil sample. Examination of the spectrum of the
peak at scan number 1054 and a library search reveals the presence of tetraethyl
lead in this particular sample (Fig. 10.13(b)). The examination of samples for
alkyl lead compounds can also be accomplished using SFE-GC with an atomic
emission detector (AED). This detector has the advantage that the detector can
selectively monitor for lead compounds in the chromatogram. This means that
the lead compounds can be detected without any interferences from hydrocar-
bons etc. which have been co-extracted. This together with the inherent sensitiv-
ity of the detector and the high concentration factors which can be achieved by
'IABLE 10.7
SINGLE ION COUNTS FOR 3- TO 6-RING PAHS OBTANED BY ON-LINE SFE-GC-MS OF
FOliR SOIL SAMPLES TAKEN FROM THE SAME SITE
Sample 3-ring 4-ring 4-ring 5-ring 6-ring

110. (M/W 178) (MIW 202) (MIW 228) (M/W 252) (M/W 276)
1 1000 2144 444 304 63

-3 595 1000 158 89 0
3 516 1373 333 345 96
4 20 32 0 0 0
Supercritical ji'uid extraction 297
569
563
l0j
0
u u
I I I I I I I I I I I
600 800 1000 1200 1400 1600 1800 2000 2200 2400 2600
Scan number
100 1 b 1065 I
1099
1054
1188
s 1048
- Scan 1054
3
LL
50
n
.7 105 'p I.
0 . I
I 1 I I I I I I I I I I
50 100 150 200 250 300 50 100 150 200 250 300
M a s s Ich a r ge Mass/charg e
Fig. 10.13. (a) Total ion chromatogram obtained from on-line SFE-GC-MS of a contaminated soil
sample. (b) Portion of total ion chromatogram shown in (a) with mass spectrum and library match
for scan number 1054 identifying the peak as tetra-ethyl lead. (Reproduced courtesy of British Pe-
troleum.)
chromatogram shows the response from the carbon emission line and the top one
the response from the lead emission line where peaks for tetramethyl and tetra-
ethyl lead can readily be detected.
In many cases mass spectrometry cannot give sufficient information to posi-
tively identify components of interest, particularly when looking for unknown
components, for example in problem-solving types of application. In these cases
"multi-hyphenated" techniques such as SFE-GC-FTIR-MS can be particularly
useful as both the infrared and mass spectrometer can be employed to give

298 Chapter I0
301 Lead tEtPb
tMePb
1 h -7
4 6 8 10 12 14 16 18 20
Time (minutes)
Fig. 10.14. Lead (top) and carbon (bottom) specific chromatograms obtained from on-line SFE-
GC-AED of a soil sample spiked with leaded gasoline. The lead concentration is equivalent to 1
part million in the soil. (Reproduced courtesy of British Petroleum.)
structural information. This allows confirmation of identification by two inde-

pendent techniques in a single experiment 1571.
SFE OVEN
I
-
EXT::f,'oN/ +
I
,
'
,
HPLC
PUMP
, ~
1
J
I DETECTOR^
fig. 10.15. Schematic representation of a coupled SFE-HPLC system employing a recirculating
atraction manifold interfaced to the HPLC via a sample injection valve.
10.5.2.2 On-line SFE-HPLC

There are many compounds which are not amenable to GC analysis and in
these cases SFE coupled on-line to liquid chromatography can be a useful tool.
The recirculating mode of SFE with an in-line sampling valve discussed earlier
provides an easy means of coupling SFE with HPLC. A schematic representa-
tion of such a coupling is shown in Fig. 10.15. In this system a small aliquot of
extract is removed from the recirculating extraction system via a sample loop.
This loop is introduced directly into the HPLC eluent where the carbon di-
oxide extraction solvent expands and passes through the HPLC system unre-
tained. The extracted components are retained and then separated on the analyti-
cal column.
A chromatogram obtained from this system for a phenol formaldehyde resin
sample is shown in Fig. 10.16. One advantage of this configuration is that the
remaining extract in the system can be collected on depressurization and can
then be examined by other techniques. In this case, off-line GC-MS and HPLC-
MS identified the peaks as phenol and isomeric phenol formaldehyde condensa-
tion products. Dynamic extraction can also be linked with HPLC and this is par-
ticularly useful where maximum sensitivity is required. One way of achieving
this is via an intermediate trap linked between the SFE and HPLC system via a
switching valve. The SF extract is expanded into an HPLC pre-column which
has been packed with the same stationary phase as the analytical column. The
extracted analytes are trapped on this column and the gaseous carbon dioxide
vented. After the extraction is complete, the HPLC eluent is introduced to the
The Ern and H P L C r n results from the isolated extract, show that the
low mlecular weight material consists of phenol and isomeric phenol
formaldehyde condensation products of the type shown below :
CHZOH HO ,OH
n
1 1 1 1 1 , 1 1 1 , , , 1 , , 1 1 , , ,
10 20 30 40 50
Time (minutes1
Fig. 10.16. Chromatogram obtained by SFE-HPLC of a phenol formaldehyde resin. The recirculat-
ing SFE system was coupled to the HPLC system as illustrated in Fig. 10.9. (Reproduced by cour-

300 Chapter I0
EXTRACTION
CELL
SmTCHlNG
/ VALMS(2)
i C02 PUMP
1
ACCUUUUTOR
/ COLUMN S
P
E
THERM0K)SPAAY C
GRADIENT T
HPLC R
PUMPS
I PARTICLE BEAM 0
M
co=Q+
IiR
Fig. 10.17. Schematic representation of possible coupled SFE-HPLC-MS systems which have
been etnployed for the examination of polymer additives and their breakdown products in polyeth-
ylene.
pre-column via a valve switch and the trapped analytes are introduced into the
HPLC system. In this case the whole extract is transferred to the HPLC system
and therefore off-line techniques for identification are not available.
However. recent developments in HPLC-MS mean that instruments are now
commercial Iy available at reasonable cost. SFE has been successfully coupled in
this way to commercial HPLC-MS systems and has been shown to give unique
information for the examination of polymer additives and their breakdown prod-
ucts [57,58]. A schematic diagram of the possible systems is shown in Fig.
10.17. Three HPLC-MS interfaces were studied namely a moving belt, a particle
beam interface, and a thermospray interface. The particle beam/thermospray
combination was particularly useful as it was available on the same instrument
and it was a simple operation to select either as required. This gave excellent
flexibility as the particle beam gives EI-type spectra and therefore fragmentation
data, and the thermospray ionization mode provides molecular weight data.
Therefore a lot of structural information on unknowns can be collected in a short
time. This system has been shown to give less degradation in polymer additives
than when an off-line SF extraction with collection in solvent was employed and
this could provide a powerful tool to study the fate of reactive compounds in
their natural environment.
10.6 CONCLUSION
The range of applications cited in this chapter has demonstrated that, even
though the analytical application of supercritical fluid extraction is a relatively
new technique when compared to the more mature areas of chromatography like
GC and LC, it is a potentially powerful tool in the armoury of the analyst work-
ing in the petroleum industry. It has advantages (sometimes unique) in terms of
both technical and environmental grounds over more conventional solvent-based
extraction processes and as the drive to remove environmentally unfriendly sol-
vents increases, the use of supercritical carbon dioxide is likely to become the
technique of choice, particularly by the Environmental Protection Agency (EPA)
in the United States.
In addition, the technology is still relatively new and is therefore currently the
subject of a great degree of research and development by instrument manufac-
turers and end users. It is likely therefore that, even in the time between writing
and publication of this chapter, many more significant developments will have
occurred in both the instrumentation and applications available to the analyst.
10.7 REFERENCES
1 R.C. Reid, J.M. Prausnitz and B.E. Poling, The Properties of Gases and Liquids, 4th edition,
McGraw-Hill; New York (1987).
2 M.A. McHugh and V.J. Krukonis, Supercritical Fluid Extraction: Principles and Practice
Buttenvorths, Boston, (1986) ISBN 0-409-90015-X.
3 T.J. Bruno and J.F. Ely (Eds.), Supercritical Fluid Technology: Reviews in Modern Theory
and Applications, CRC Press, Boca Raton, FL (1991) ISBN 0-8493-6847-2.
4 M.L. Lee and K.E. Markides (Eds.), Analytical Supercritical Fluid Chromatography and Ex-
traction, Chromatography Conferences (1990) ISBN 0-8425-2394-4.
5 F.V. Bright and M.E.P. McNally (Eds.), Supercritical Fluid Technology: Theoretical and
Applied Approaches in Analytical Chemistry. ACS Symposium Series 488 (1992) ISBN 0-
8412-2220-7.
6 R.M. Smith (Ed.), Supercritical Fluid Chromatography. RSC Chromatography Monographs
ISBN 0-85186-577-1.
7 E. Klesper, A.H. Convin and D.A. Turner, J. Org. Chem., 27 (1962) 700.
8 S.T. Sie, W. van Beersum and G.W.A. Rijnders, Separation Sci., 1 (1966) 469.
9 S.T. Sie and G.W.A. Rijnders, Separation Sci., 2 (1967) 699.
10 Sie, S T; Rijnders, G W A. Sep Sci, Vol2, 729, 1967
11 Sie, S T; Rijnders, G W A. Sep Sci, Vol2, 755, 1967
12 Sie, S T; Rijnders, G W A. Anal Chim Acta, Vol38, 31, 1967.
13 R.E. Jentoft and T.H. Gouw, J. Chromatogr. Sci., 8 (1970) 138.
14 R.E. Jentoft and T.H. Gouw, J. Polym. Sci., Polym. Lett., 7 (1969) 81 1.
15 J.R. Combes, K.P. Johnston, K.E. OShea and M.A. Fox, Supercritical Fluid Technology:
Theoretical and Applied Approaches in Analytical Chemistry, Chapter 3, ACS Symposium
Series 488 (1992) ISBN 0-8412-2220-7.
302 Chapter I0
16 D.R. Gere, R. Board and D. McGinigill, Anal. Chem., 54 (1982) 736.

17 T. Greibokk, AL. Blillie, E.J. Johansen and E. Lundanes, Anal. Chem., 56 (1984) 2968.
18 J.M. Levy, Advances in Analytical SFE, American Laboratory, August (1991).
19 K.H. Pickel? Proceedings of the 2nd International Symposium on Supercritical Fluids, Boston
(1991) p. 457.
20 M. Saito and Y. Yamauchi, Supercritical Fluid Extraction and Supercritical Fluid Chromatog-
raphy for Preparative Separation, Jasco Report Publisher, Japan (1990).
21 B.W. Wright, C.W. Wright, R.W. Gale and R.D. Smith, Anal. Chem., 59 (1987) 38.
22 H.M. McNair and J.O. Frazier, American Laboratory (1991) 24d.
23 L.J. Mulcahey, J. Hedrick and L.T. Taylor, Anal. Chem., 63 (1991) 2225.
24 M. Ashraf-Khorassani, R.K. Houck and J.M. Levy, J. Chromatogr. Sci., 30 (1992) 361.
25 N.L. Porter, A.F. Ryanski, E.R. Campbell, M. Saunders, B.E. Richter, J.T. Swanson, R.B.
Nielsen and B.J. Murphy, J. Chromatogr. Sci., 30 (1992) 367.
26 J.L. Hedrick and L.T. Taylor, Anal. Chem., 61 (1989) 1986.
27 J.L. Hedrick, L.J. Mulcahey and L.T. Taylor, Supercritical Fluid Technology: Theoretical and
Applied Approaches in Analytical Chemistry, Chapter 15 (F.V. Bright and M.E.P. McNally,
Eds.), ACS Symposium Series 488 (1992) ISBN 0-8412-2220-7.
28 Supercritical Fluid Technology Synopsis SFE Application: PCBs in Oil. SFE-82 (1991),
Suprex Corporation. Pittsburgh, PA.
79 D.C. White, D.E. Nivens, D. Ringelberg and D. Hedrick, Proceedings of the International
Symposium on Supercritical Fluid Chromatography and Extraction, Park City, UT (1991) p.
43.
30 S.B. Hawthorne, D.J. Miller, D.E. Nivens and D.C. White, Anal. Chem., 64 (1992) 405.
31 D.J. Miller, S.B. Hawthorne, J.J. Lagenfield and D.C. White, Proceedings of the International
Symposium on Supercritical Fluid Chromatography and Extraction, Park City, UT (1991). p.
155.
32 J.J. Hills, H.H. Hill and T. Maeda, Proceedings of the International Symposium on Supercriti-
cal Fluid Chromatography and Extraction, Park City, UT (1991) p. 113.
33 J.J. Hills. H.H. Hill and T. Maeda, Anal. Chem., 63 (1991) 2152.
34 C.P. Eppig, B.M. Putnam and R.P. de Fillippi, US Patent application no. 255,037, filed April
17th (1981).
35 Supercritical Fluid Technology Synopsis, SFE-87, Suprex Corporation, Pittsburgh, PA
(1991).
36 Off-line supercritical fluid extraction with the Dionex SFE-703, Dionex Corporation, Salt
Lake Division.
37 B. Subramaniam and S. Saim, J. Supercritical Fluids, 3 (1990) 214.
38 S. Saim , D.M. Ginosar and B. Subramaniam, Proceedings of the 2nd International Sympo-
sium on Supercritical Fluids, Boston, MA (1991) p. 137.
39 S.A. Schmidt, R.E. Yoder and J.V. Smith, Supercritical Fluid Extraction of Refinery Bottom
Sludges and the Analysis for Polynuclear Aromatic Hydrocarbons, Application Note 228-1 19.
November 1990, Hewlett-Packard Company.
40 G.W. Kelly, K.D. Bartle and A.A. Clifford, Proceedings of the 4th International Symposium
on Supercritical Fluid Chromatography and Extraction, Cincinnati, OH (1992) p. 139.
41 R. Flaherty and J.V. Smith, Hewlett-Packard application note 228-133, Supercritical Fluid
Extraction of Filter Media. April 1991.
42 A. Lopez-Avilla, J. Benedicto. N.S. Dodhiwala and R. Young, J. Chromatogr. Sci., 30 (1992)
335.
Supercriticalfluid extraction 303
43 Determination of Total Petroleum Hydrocarbons (TPH) in Soil, Application Note Ref RC-5,
Suprex Corporation, Pittsburgh, PA (1991).
44 M. Richards and R.M. Campbell, LC-GC Int., 4 (1991) 33.
45 N.J. Cotton, K.D. Bartle, A.A. Clifford, S. Ashraf, R. Moulder and C.J. Dowle, J. High Reso-
lut. Chromatogr., 14 (1991) 164.
46 H. Daimon and Y. Hirata, Chromatographia, 32 (1991) 549.
47 M. Ashraf-Khorassani, D.S. Boyer and J.M. Levy, J. Chromatogr. Sci., 29 (1991) 517.
48 R.W. Wright, S.R. Frye, D.G. McMinn, R.D. Smith, Anal. Chem., 59 (1987) 640.
49 J.C. Monin, D, Barth, M. Perrut, M. Espitalie and B. Durand, Report Ref 35604, Institute
Francais du Petrole (1987).
50 S.B. Hawthorne and D.J. Miller, J. Chromatogr. Sci., 403 (1987) 63.
51 S.B. Hawthorne, D.J. Miller and M.S. Kreiger, J. Chromatogr. Sci., 27 (1989) 347.
52 S.B. Hawthorne, D.J. Miller and M.S. Kreiger, Anal. Chem., 60 (1988) 472.
53 S.B. Hawthorne, D.J. Miller, M.S. Kreiger, Fresenius Z. Anal. Chem., 330 (1988) 21 1.
54 J.M. Levy, J.P. Guzowski, W.E. Huhak, J. High Resolut. Chromatogr., Chromatogr. Com-
mun., 10 (1987) 337.
55 J.M. Levy and J.P. Guzowski, Fresenius Z. Anal. Chem., 330 (1986) 207.
56 J.M. Levy and A.C. Roselli, Chromatographia, 28 (1986) 613.
57 T.P. Lynch, R.E.A. Escott, P.G. McDowell, I. Roberts and M.J. Carrott, Proceedings of the
4th International Symposium of Supercritical Fluid Chromatography and Extraction, Cincin-
nati, OH (1992) p. 179.
58 T.P. Lynch, Applications of Supercritical Fluids in Industrial Analysis (J.R. Dean, Ed.),
Blackie, Glasgow (1993) Chapter 8, p. 208, ISBN 0-7514-057-2.
Journal of ChromatographyLibmy Series, Vol. 56
CHAPTER 11
Supercritical fluid chromatography
I. Roberts
Analytical & Applied Science Division, BP Research & Engineering Centre, Sunbury-on-Thames, Middlesex,
TWl6 7LN UK
11.1 INTRODUCTION
The term supercritical fluid chromatography (SFC) was introduced by Sie and
Rijnders in 1967 [I], some 9 years after Lovelock had first proposed using su-
percritical carrier gases for the analysis of non-volatile compounds by gas chro-
matography (GC). In 1962 the first published SFC application appeared de-
scribing the separation of porphyrins on a packed GC column using supercritical
chlorofluoroalkanes as the mobile phase [2].
The petroleum industry immediately recognised the possibilities offered by
this new form of chromatography. Sie and Rijinders carried out much of the
early pioneering work at the Shell Laboratories in Amsterdam [3,4] while, at
Chevron, Jentoft and Gouw demonstrated the use of polar modifiers and pressure
programming as a means of controlling retention in SFC [5,6]. Although work
on SFC continued during the 1970s, particularly at Chevron [7], the oil industry
lost interest in SFC as it focussed its attention on the emerging technique of high
performance liquid chromatography (LC).
In an earlier review, Gouw and Jentoft described the status of SFC in petro-
leum analysis as it stood in 1978 [8]. Separations were performed on packed
columns containing large particle-sized stationary phases, detection was pre-
dominantly by UV and typical mobile phases were alkanes (e.g. pentane, hex-
ane) and oxygenated solvents (e.g. propan-2-01, diethyl ether). Although carbon
dioxide was in use as a mobile phase, it did not occupy the dominant position it
does today. Open tubular capillary columns had not yet arrived on the scene and
instrumentation was still built in-house. The GC origins of SFC were clearly
evident in that the main benefits of SFC were perceived to be the separation of

306 Chapter I I
components with low GC migration rates and/or low thermal stabilities. Inter-
estingly, this review did not mention supercritical fluid extraction (SFE) but did
give an example of SFC directly coupled to mass spectrometry (SFC-MS).
The beginning of the 1980s saw renewed interest in SFC spurred on by the
development of open tubular capillary columns [9] and the introduction of the
first commercial SFC instrument, a modified Hewlett-Packard liquid chroma-
tograph, in 1982. In 1986 the first commercially-built capillary supercritical fluid
chromatograph appeared. This period of rapid growth, up to the present day, has
seen many improvements in instrumentation, particularly injector and detector
technology, and on-line coupling with SFE, GC, LC, MS and infrared (IR).
Capillary and packed columns are now more robust and more efficient while
developments in the design and synthesis of stationary phases has produced op-
timised SFC phases with a range selectivities. The theoretical basis of SFC is
now well established [ 101.
In spite of the technical success of SFC, there has been a reticence within the
general scientific community to accept it as an equal partner with GC and LC.
Some of the blame for this must be attached to exaggerated claims, made in the
early 1980s, that SFC would make GC and LC redundant. Perhaps there was too
much emphasis on the theoretical aspects, during these early development years,
at the expense of demonstrating the problem-solving powers of SFC (cf. the de-
velopment of GC and LC). Certainly a large number of the published SFC appli-
cations dealt with analytical solutions which were more readily achievable by
GC or LC, and encouraged the sceptics to dismiss SFC as a technique looking
for an application. Nowadays SFC is rightly regarded as a complementary, if
still minor, partner in the GC/SFC/LC triad of column chromatography.
The modern revival in petroleum SFC may be traced to a paper by Rawdon,
published early in 1984, describing a modified flame ionization detector (FID)
for packed columns [I I]. The significance of this paper was that petroleum ana-
lysts could now perform quantitative hydrocarbon-type analyses which had pre-
viously proved elusive using LC techniques. The example Rawdon gave in his
paper was the separation of diesel fuel into saturates and aromatics; some 9 years
later the first standard SFC test method (ASTM D5186-91: Determination of
Aromatic Content of Diesel Fuels by Supercritical Fluid Chromatography) was
published [ 121.
11.2 INSTRUMENTATION
The supercritical fluid chromatograph is a hybrid instrument assembled from GC

and LC components. The basic elements of the SFC instrument are illustrated
schematically in Fig. 1 1 . 1 . Mobile phase, in the form of a liquid or pressurized
Supercriticalfluidchromatography 307
detector X )
1
-_- I
0
r-----
i-
restrictor
detector LC)
_ - _ __ _ _ _ _
-----a
restrictor
OVEN
co2
Fig. l l .1. Schemata of SFC chromatography.
gas, is supplied to the LC pump where the pressure of the fluid is raised above
its critical pressure. Samples are introduced into the fluid stream via an LC in-
jection valve and separated on a column placed in a GC oven thermostatted
above the critical temperature of the mobile phase. A post-column restrictor en-
sures the fluid is maintained above its critical pressure throughout the separation
process. Analytes eluting from the column are monitored by detectors positioned
either before or after the post-column restrictor.
Prior to the mid-l980s, laboratories designed and assembled home-made SF
chromatographs using readily available LC (pumps, injectors, columns and de-
tectors) and GC (ovens, columns, detectors) components. The modifications re-
quired to build a working SFC instrument are relatively minor so that this ap-
proach can still provide a cheap introduction to SFC by using redundant LC and
GC parts. For industrial laboratories, it is more cost-effective to invest in pur-
pose-built instrumentation which is now available from a number of suppliers
and manufacturers. Commercial SF chromatographs offer sophisticated micro-
processor-controlled units (allowing the simultaneous programming of fluid
pressure, temperature, flow and composition), a range of detector options, auto-
mated sample injection and on-line coupling with SFE. Of equal importance is
the back-up (maintenance, spare parts, consumables and practical advice) pro-
vided by the manufacturers.
11.2.1 Mobile phase pumps
The specifications for an SFC pumping system are similar to those for LC, i.e. it
should provide a stable pulseless flow over the operating pressure range

308 Chapter 11
(typically 70-400 bar). Pumps designed to operate at higher pressures (e.g. up to

650 bar) are available but their use is often restricted by pressure limitations on
other system components such as injection valves. Since flow rate requirements
vary from a few pl min-I in capillary SFC to several ml min-l in packed column
SFC, two different pump types are used to cover this range.
11.2.1.1Reciprocatingpumps
Generally the cheapest option for SFC, these conventional LC pumps supply
stable flow rates over the range 0.5-5 ml m i d , making them ideal for use with
packed columns having internal diameters >2 mm. For efficient pumping, the
pump heads must be cooled to 0C or less by means of a clamp-on heat ex-
changer and a recirculating chiller. Stream-splitting may be used to reduce the
flow rate but this is not a reliable means for producing the very low flow rates
required for capillary SFC unless flow and pressure can be precisely and accu-
rately maintained through a programmable, electronically-controlled variable
restrictor [ 131.
A major advantage of reciprocating pumps is the ease with which liquid
modifiers can be added to the main fluid using an additional pump or pumps (the
modifier pump does not need to chilled). Standard high pressure LC mixers en-
sure rapid and reproducible mixing of the mobile phase streams so that compo-
sitional gradients can be used to enhance selectivity or elute components of
widely varying polarity.
The check valves in the chilled pump head need to be replaced more fre-
quently than their LC counterparts but, otherwise, reciprocating pumps offer low
maintenance and trouble-free operation.
11.2.1.2 Syringepumps
The ability of a syringe pump to provide a virtually pulse-free flow in the 1-
10 pl min-' range makes them ideal for capillary SFC. They are also suitable for
use with the larger diameter packed columns provided the capacity of the syringe
(typically 20-500 ml) is adequate.
Syringe pumps are frequently used with packed columns up to 1 mm ID; pis-
ton seal wear can become significant if flow rates of several ml min-' are used
routinely.
Efficient filling of the pump is achieved either by cooling the pump cylinder
to around 5C or by using gas cylinders in which the mobile phase is overpres-
surized with helium at 100-125 bar. The former method is preferred since the
presence of even small amounts of helium in the mobile phase can have a detri-
mental effect on chromatographic reproducibility in capillary SFC [ 141.
The fundamental design of syringe pumps (large cylinder volumes and no
check valves) combined with high fluid compressibilities gives rise to a major
Supercritical fluid chromatography 3 09
drawback, the inability to generate mobile phase composition gradients by add-

ing a liquid (low compressibility)modifier to the highly compressible fluid [15].
Additionally, the least compressible liquid may force its way back into the tub-
ingkylinder of the highly compressible fluid giving rise to cross contamination
problems and inaccurate or variable solvent compositions.
A single syringe pump can be used to deliver a mixed solvent, under isocratic
conditions, either by using cylinder bottles pre-doped with modifier (e.g. 10%
m/m methanol in carbon dioxide) or by pre-mixing the liquid modifier and fluid
in the pump cylinder [16,17]. The practical difficulties in generating mixed
mobile phases with syringe pumps are only a minor inconvenience for the petro-
leum analyst where multicomponent fluids are the exception rather than the rule.
Modern syringe pumps, although more expensive, are extremely reliable and
easy to maintain. The piston drive mechanism requires regular lubrication to
prevent corrosion problems arising from water condensation (due to cylinder
cooling) and the high pressure piston seal needs to be replaced approximately
every 6 months.
11.2.2 Ovens/temperature control
The role of the oven in SFC is to raise and maintain the temperature of the mo-
bile phase, in a controlled and repeatable manner, above its critical temperature.
Standard GC ovens meet all the requirements for temperature management in
SFC by providing accurate, precise and uniform (no localized hot or cold spots)
control of temperatures, including temperature programming, from ambient to
300C. Most commercial SFC instruments are supplied with GC-type ovens so
that they can also operate in the GC mode. Many home-made SF chroma-
tographs utilize redundant GC ovens, not only for their temperature control
function, but because they have additional in-built features such as detectors,
heated injectioddetection zones and a large oven cavity. The temperature pro-
gramming facility, although not widely used in petroleum SFC applications, is
an area worthy of further investigation.
A number of cheaper options are available for packed column SFC carried out
under isothermal conditions (the most common requirement for petroleum SFC
applications). Circulating air ovens and block column heaters designed for LC
provide excellent temperature control and even a simple oil or water bath is ade-
quate.
11.2.3 Injectors
Sample introduction is often the most critical part of any chromatographic sys-
tem. The injection system aims to transfer a concentrated (minimum band

310 Chapter 11
spreading), representative (i.e. no sample discrimination) and repeatable (i.e.

quantitative) aliquot of the sample to the head of the column without disturbing
the chromatographic equilibrium. In practice there is no ideal injection mecha-
nism for SFC and the injection mode that works best for one sample does not
necessarily work best for others.
The standard LC injection valve has monopolized the introduction of liquid
samples (including solutions) onto both capillary and packed columns, even
though the amount of sample injected onto a packed column (e.g. 1Opl) may be
three orders of magnitude greater than that injected onto an open tubular capil-
lary column (e.g. 10 nl). Commercial LC injectors offer fixed volume injections
from 60nl upwards, allowing the hi1 loop volume to be injected onto packed
columns (>OS mm ID) with excellent repeatability (4% RSD) [15]. Sample
injection onto packed columns presents no real problems provided the analyst
follows standard chromatographic practices.
For capillary columns, injection volumes smaller than 60 nl are preferred and
various methods have been devised to achieve this. The dynamic split injection
method uses a two-way splitter, positioned between the injector and column, to
send most of the sample (e.g. 99%) to waste and only a small aliquot onto the
column. Timed-split injection relies on the ability to switch the injection valve
from load to inject and back to load over very short time periods (tens of milli-
seconds) so that only a portion of the loop volume is pushed on to the column by
the mobile phase. Both methods are available on commercial instruments and are
capable of good repeatability, especially if internal standards are used. Overall,
timed split injection has a slight edge over dynamic split by virtue of its better
repeatability, lower sample discrimination, reduced solvent tailing and more
flexible adjustment of injection volume.
A disadvantage of the split injection techniques is that complex samples, of
which petroleum is a prime example, must be injected as concentrated solutions
because of the very small volumes transferred to the column. Preparing concen-
trated solutions may require the use of either a strong solvent, which may not
be compatible with the mobile or stationary phase, or heating the solution to im-
prove solubility. In the latter case, the injection valve must also be heated to en-
sure the sample does not precipitate in the valve and cause problems with sample
discrimination and carry-over. A number of solvent elimination (or venting)
techniques have been developed to allow the injection of more dilute sample
solutions. A fuller discussion of these techniques, which bring trace analysis into
the realm of capillary SFC, may be found elsewhere [ 15,181.
11.2.4 Detectors
One of the major attractions of SFC is the ability to mix and match detectors
Supercriticaljluid chromatography 311
which hitherto were only available exclusively to GC or LC. Most GC and LC

detectors can be used with little or no modification on both packed and capillary
column systems. LC detectors normally require a high pressure flow cell for use
with packed columns and a degree of miniaturization for capillary SFC. GC de-
tectors can be used without modification on capillary systems but some allow-
ance for the higher flow rates must be made for packed column SFC applica-
tions. In column chromatography there are two basic detector designs: (i) liquid
phase detectors where the analyte remains in solution, and (ii) gas phase detec-
tors where the analyte is vaporized. In SFC the former (usually LC detectors) are
located before the restrictor and the latter (mainly GC detectors) are positioned
after the restrictor.
I 1.2.4.1 Universal detectors

Simple detectors whose primary function is to sense all analytes eluting from
the chromatographic column are termed universal detectors. They provide mini-
mal chemical or structural information on the eluate but are routinely used for
quantification. More sophisticated instruments, such as mass spectrometers and
Fourier transform infrared spectrophotometers, can also function as universal
detectors although, for reasons of cost, this is not their primary role.
(i) Flame ionization detector (FID).The FID is the most popular detector in SFC
due, at least in part, to its widespread use for petroleum analysis [19]. As the
first-choice detector for quantitative analysis of petroleum hydrocarbons it has
played a major role in the development of SFC within the oil industry. Apart
from optimizing the flame gas conditions [20], the standard FID can be used
without modification on both packed and capillary columns. Large bore packed
columns (>2 mm ID) require post-column flow splitting to avoid extinguishing
the flame [11,21]. With carbon dioxide as the mobile phase, FID sensitivity and
linearity are comparable to those achieved in GC.
Nitrous oxide, sulphur hexafluoride and xenon are also compatible with the
FID. Nitrous oxide produces a much higher background signal than carbon diox-
ide, giving a sensitivity reduction of several orders of magnitude. Nickel- or
gold-plated collectors must be used with sulphur hexafluoride to counteract the
corrosive action of hydrogen fluoride formed in the FID flame. Xenon is used
exclusively with flow cell IR detectors, for reasons of cost, but may subse-
quently be directed to an FID for quantification.
The sensitive and universal response of FIDs to organic compounds effec-
tively precludes the use of organic modifiers with carbon dioxide or other FID-
compatible fluids. Formic acid and water, which are sparingly soluble (4%) in
carbon dioxide, are the only two polar modifiers which have been used success-

312 Chapter 11
fully with the FID [22,23]. Alternative detectors must be found when other or-
ganic modifiers (e.g. methanol) are used.
(ii) Photo-ionization detector (PID). The PID is a non-destructive GC detector

often placed in series with an FID and with which it shares a number of useful
characteristics (responsive to most organic compounds, high sensitivity and wide
linear dynamic range). In other respects, however, the PID (concentration- and
compound-sensitive response) and FID (mass-sensitive and uniform hydrocarbon
response) are complementary to each other.
In the PID, a sealed excitation source emits photons in the far UV which
ionizes analytes having an ionization potential (IP) less than the energy of the
emitted photons. The IPS of carbon dioxide, nitrous oxide and sulphur hexafluo-
ride are 13.79 eV, 12.89 eV and 19.3 eV, respectively, so that these fluids should
be transparent to the PID when using the 10.2 eV excitation source. Some of
the common SFC modifiers also have high IPS, e.g. methanol (10.85 eV)
dichloromethane ( I 1.35 eV) and acetonitrile (12.22 eV), suggesting the possi-
bility of using mixed fluids with this detector. In practice, the addition of
even low concentrations of methanol modifier significantly reduces the sensitiv-
ity of the PID and there have been few published applications of its use in
SFC.
(iii) Light scattering detector (LSD). If the LSD had not originally been devel-
oped for LC, then it would almost certainly have been developed for packed col-
umn SFC. The detection mechanism is complex but, in simple terms, relies on
the nebulization of the column effluent to form droplets and particles which
cause light to be scattered as they pass through a light beam. The amount of
scattered light is measured and is a non-linear function of the analyte mass
passing through the detector.
For LC, an inert gas must be used to nebulize the liquid eluent whereas for
most SFC applications the natural expansion of the supercritical fluid provides
efficient nebulization [24,25]. In the nebulization process, volatile components
(including organic modifiers) do not form droplets or particles and are therefore
not detected by the LSD. The LSD is therefore unaffected by composition and
pressureldensity gradient programming and makes an excellent universal detec-
tor suitable for use with all common SFC mobile phases. The sensitivity of the
detector (low ng) is adequate for most purposes. The only drawback is that the
detector response, in LC and SFC modes, is compound-dependent and non-
linear. For the analysis of homologues (e.g. polymers) or individual species, this
drawback is easily overcome by appropriate calibration of the detector. How-
ever, many petroleum analyses determine hydrocarbon types or compound
classes for which there are no standards available for calibration. The composi-
Supercritical fluid chromatography 313
T = 90C
P = 135 to 285 bar at 2.27 bar/min
0 5 10 I5 20 25 30 35 40 45 50 55 60 65 710
minutes
T = 80%
P = 160 to 250 bar at 5.5 bar/min
5
4 1
I I I 1 I I
10 15 20 25 30 ;f,
minutes
Fig. 11.2. SFC anslysis of a diesel fuel with (a) FID and (b) UVD at 254nm detection. Mi-
cropacked fused silica column (300 mm x 0.32pm ID), C02 mobile phase. Peak identification: (1)
naphthalene; (2) methylnaphthalene; (3) fluorene; (4) phenanthene; (5) methylphenmthrene
(reproduced from ref. 25 with permission of Preston Publications).

3 14 Chapter I I
tional variation within a hydrocarbon-type fraction (e.g. aromatics) even pre-

cludes the use of such fractions themselves as calibration standards.
11.2.4.2Spectroscopic detectors
Compounds possessing certain structural features (e.g. an aromatic ring or car-
bony1 group) can be selectively identified in the column effluent using spectro-
scopic detectors. The ubiquitous UV/visible detector (UVD) currently vies with
the FID as the most popular general-purpose SFC detector, although this position
is largely due to its use on packed column SFC systems and non-petroleum ap-
plications.
(9 UV/visible detector (UVD). Many compounds possess a W-absorbing func-

tional group in the 200-800 nm spectral region and are readily detected with
high sensitivity by the UVD. The majority of compounds found in petroleum are,
however, saturated hydrocarbons which do not respond to the UVD. Since the
UVD is non-destructive, this limitation can be turned to advantage by using the
selectivity of the UVD to monitor olefins and aromatics in combination with the
universal detection capabilities of the FID. Figure 1 1.2 illustrates the potential of
selective UVD detection for the SFC analysis of diesel fuel. The SFC-FID chro-
matogram (Fig. 1 1.2 (a)) is dominated by the saturated hydrocarbons (especially
the normal paraffins) yet it is still possible to identify the underlying polycyclic
aromatic hydrocarbons (PAHs) in a single run using the UVD (Fig. 11.2(b)).
The UVD is easily interfaced to packed column SFC instruments; installation
of a high pressure flow cell is usually the only modification required. Capillary
SFC places stringent limitations on the detector cell volume which are usually
met using on-column or pseudo on-column flow cells [27]. The use of Z-shaped
flow cells (Fig. 11.3), now commercially available, allows the cell path length
(and therefore UV response) to be increased dramatically although at the ex-
pense of higher noise levels [28]. Photodiode array detectors have been used
with both packed [29] and capillary columns [30], although the latter suffers
from excessively high noise levels; rapid scanning W D s offer improved signal
to noise ratios for capillary systems [3 11.
The common supercritical fluids are useable over the full W/visible spec-
trum, as are n o n - W absorbing modifiers (e.g. methanol, acetonitrile). High pu-
rity carbon dioxide can be used below 200 nm which enables its use as an olefin-
selective detector for hydrocarbon-type analysis. Baseline drift observed with
pressure/density programming has been attributed to density-related refractive
index changes; this effect is minimized if detection is performed under subcriti-
cal (i.e. liquid state) conditions.
(ii) Fluorescence detector (FLD). For certain compounds (e.g. PAHs), consid-
erably lower limits of detection can be achieved using a FLD in place of a UVD.
lamp tubing photo diode
On-column: path length 75 pm
Z-shape: path length up to 20 mm
Fig. 11.3. Comparison of on-column and Z-shaped flow cell design for UV/visible detectors
(reproduced with permission of LC Packings).
The extra sensitivity, coupled with a high degree of selectivity, makes fluores-
cence detection a particularly attractive option for capillary SFC where the flow
cell is often an integral part of the column itself [32]. A more recent develop-
ment is laser-induced fluorescence/supersonic jet spectroscopy (LIFEJS) where
the effluent from the column expands into a low pressure region to produce a
narrow beam of cold gas phase molecules. The very high resolution spectra
obtained under these conditions enables compounds, even geometric isomers, to
be accurately identified. At present the sensitivity of the technique, even with
LIF, is insufficient for real time identification of chromatographic peaks [33].
Virtually all supercritical fluids are compatible with the FLD as are density
programming and compositional mobile phase gradients. The following mobile
phases have been used: carbon dioxide (with and without modifiers), alkanes
(propane, butane, pentane), dichlorofluoromethane, and ammonia. Fluorescence
detection, with the exception of SJS, is non-destructive and hence may be used
in conjunction with other detection methods.

316 Chapter I 1
(iii) Mass spectrometry detector (MSD). MSD is the most popular detection
method after FID and UVD for the following reasons: high sensitivity (low pg)
and selectivity (especially with MS-MS techniques), plus the ability to identify
or confirm unknown analytes. For routine oil laboratories, the costs of purchas-
ing and maintaining an SFC-MSD system may not be justifiable. Nevertheless
this is an area of considerable interest to the petroleum analyst because of the
largely unknown composition of petroleum. The sensitivity/selectivity combina-
tion of mass spectrometry enables more complex mixtures to be analysed thus
reducing or minimizing sample preparation times. Higher selectivities (with
lower sensitivities) are possible with high resolution magnetic sector instruments
or MS-MS techniques. Cheaper benchtop quadrupole instruments are, however,
often adequate for most analyses where the SFC separation has removed possible
interferents. Facilities for both electron impact (for structural identification) and
chemical ionization (molecular mass information) are desirable but not always
attainable with the same SFC-MSD interface.
Prior developments in GC-MS and LC-MS interfaces have greatly accelerated
the introduction of commercial SFC-MSD systems. Since mass spectrometers
operate under high vacuum, they can only accept small gas flows such as those
generated by open tubular capillary columns. SFC-MSD interfaces for capillary
columns are therefore the simplest and use some form of direct fluid injection
(DFI) where the commonest problems are restrictor blockage and loss in sensi-
tivity with density programming (due to increased ion-source pressures). The
higher flow rates generated by packed columns can still be accommodated by a
DFI interface if only a portion of the column effluent is sent to the mass spec-
trometer. This strategy, however, gives rise to a large loss in sensitivity and there
are a number of alternative interfaces (e.g. thermospray, moving belt, particle
beam and atmospheric pressure ionization (MI))which can take the full flow
(1-2 ml/min) from a packed SFC column. SFC-MSD interfaces have been re-
viewed by Lee and Markides [ 151.
(iv) Infrared detector (IRD). The strength of infrared spectroscopy lies in its
ability to characterize functional groups (e.g. carbonyl groups in acids, esters and
amides) and differentiate between isomeric compounds. Its major weakness,
particularly with respect to petroleum analysis, is that it is poor at identifying
homologues; fortunately, information on homologous compounds can be readily
obtained via the chromatographic separation andor by interfacing to an MSD.
Conventional dispersive infrared spectrophotometers can only be used as chro-
matography detectors if fixed at a single wavelength. To obtain spectral infor-
mation on-the-fly it is necessary to use Fourier transform IR (FTIR) instruments.
SFC-IRD interfaces may be of the flow cell type (on-line detection) or use
solvent elimination techniques (off-line detection). Early designs used high pres-
Supercriticalfluid chromatography 317
sure flow cells to enable real time non-destructive monitoring of the column elu-
ent. Chromatograms are reconstructed using the Gram-Schmidt algorithm which
has the added benefits of removing the baseline drift caused by density/pressure
programming and increasing signal intensity. The IR spectra for each chroma-
tographic peak can be extracted for identification purposes; exact matches with
library spectra may not always be possible due to changes in the position and
intensity of the IR absorption bands arising from solute-solvent interactions.
Both packed and capillary SFC systems can be interfaced to the FTIR spectro-
photometer although the difficulties are considerably greater for capillary sys-
tems where small cell volumes dramatically reduce sensitivity (stopped flow
techniques can be used to increase sensitivity). The choice of mobile phase is
fairly restrictive for on-line detection because very few solvents are transparent
over the whole infrared region. Carbon dioxide is, fortunately, one of the better
fluids although it is only transparent over the 800-2100 cm-l and 2500-
3500 cm-1 region of the IR spectrum; organic modifiers cannot be used. Super-
critical xenon is an excellent mobile phase and completely transparent over the
whole IR spectrum but is very expensive. The separation of a synthetic mixture
0 10 Time/ min 40 I
t
90
I
Pressure/atm
I
310
I I 1
4000 2000 700
cm-1
Fig. 11.4. Capillary SFC-FTIR chromatogram of model PAHs using xenon mobile phase: (a) re-
constructed chromatogram from total IR signal; (b) IR spectra of peaks 1 { chrysene) and 2 (mixture
of 9,10-diphenylanthracene,2a, and perylene, 2b) (reproduced from ref. 34 with permission of
Elsevier Science Publishers).

318 Chapter I I
of PAHs by capillary SFC-IRD using xenon mobile phase and a low volume IR
flow cell (sub-microlitre) is illustrated in Fig. 11.4 along with examples of the
high quality IR spectra which can be obtained [34].
Eliminating the solvent removes any limitations arising from the choice of
mobile phase and can also provide a large enhancement in sensitivity. The col-
umn restrictor is placed close to a moving window of IR transparent material,
KBr or ZnSe for transmission mode or aluminium for reflectance mode [35], and
the analytes deposited as a discrete spot as the mobile phase evaporates. The
moving window is cooled to minimize spot size and improve recovery of volatile
components. FTIR measurements can be made on-line (real time chroma-
tograms) or off-line (highest sensitivity by repeatedly analysing the spot) using
an FTIR microscope. A more detailed review of SFC-IRD interfaces can be
found elsewhere [36].
(19 Nuclear magnetic resonance i.R) detection. Allen and co-workers have
reported the direct coupling of SFC with a proton (H) nuclear magnetic reso-
nance spectrometer [37]. Using a packed column (250 mm X 4.6 mm ID) and
carbon dioxide modified with 1% deuteroacetonitrile as mobile phase, these
workers were able to obtain reasonable on-line proton spectra for a series of
model hydrocarbons. Proton N M R is widely used throughout the petroleum in-
dustry for characterizing saturated and aromatic hydrocarbons after chroma-
tographic separation.
11.2.4.3Element specific detection
Element specific detectors are widely used in the petroleum industry for the GC
analysis of volatile non-hydrocarbons, particularly those containing nitrogen and
sulphur. The FID is often described as a carbon detector. Element and analyte
specific detection is very useful for trace analysis and can eliminate the need for
extensive sample clean-up.
11.2 4.3.1 Thermionic detectors. The thermionic detector (TID) is used for the
selective determination of nitrogen- and phosphorus-containing compounds and
is often referred to as a nitrogen-phosphorus detector (NPD). Organo-nitrogen
and -phosphorus compounds are combusted in a hydrogenlair flame to form
electronegative specieshons which react with hot alkali metals (rubidium or
caesium), supported on a glass or ceramic bead, to produce a current which is
amplified to give the TID signal. When operated in an inert atmosphere (i.e. ni-
trogen rather than hydrogenlair) the TID exhibits high selectivity for nitro-
compounds [38].
Both carbon dioxide and nitrous oxide have been used as mobile phases
[38,39] although there may be some loss in nitrogen selectivity and sensitivity
when nitrous oxide is used. Best results are obtained by optimizing the detector
parameters (e.g. gas flows, bias voltage) for the chosen mobile phase and type of
analyte; even so the detection limit and specificity are likely to be less than that
achievable by GC. The influence of organic modifiers on the TID response ap-
pears to be variable; the TID operated successfully with 7% methanol in carbon
dioxide [23] but not with methanol concentrations higher than 0.8% in nitrous
oxide [40].
11.2.4.3.2 Flame photometric detection. The flame photometric detector (FPD)

offers selective detection for organosulphur and organophosphorus compounds
by measuring the chemiluminescence emitted as excited S2 dimers and HPO
species, generated in the hydrogedair (or hydrogedoxygen) ffame, return to the
ground state. Quantification with the FPD, in GC and SFC, is not trivial because
of its non-linear response, which is compound dependent, and quenching of the
sulphur signal by co-eluting hydrocarbons. The dual flame photometric detector
(DFPD) is better but still far from ideal.
Carbon dioxide is the only reported mobile phase used with the FPD.
Markides et al. found detection limits for sulphur (as benzo[b]thiophene) in the
low ng range and a rapidly increasing baseline as the mobile phase pressure was
increased [41]. Olesik et al. studied the flame gas chemistry in SFC, using ex-
perimental design algorithms, and found that the optimum gas flow rates for sul-
phur detection were very different from those used in GC [20]. Detection limits
under these optimum conditions approached those obtained in GC and baseline
drift with increasing pressure was not observed when a tapered restrictor, rather
the linear restrictor used by Markides et al., was employed.
11.2.4.3.3Sulphur chemiluminescence detection (SCD). There are three types of

SCDs: fluorine-induced SCD, redox CD and the ozone-induced SCD. These de-
tectors work by first oxidizing the organosulphur compound to give a species
which may either react with fluorine or ozone to form a chemiluminescence
species (HF*and SO2*,respectively) or in the case of the redox CD it is the NO,
produced from the oxidation of the sulphur compound by NO2, which is reacted
with ozone to produce the chemiluminescent NO2*.
The FSCD, like the FPD, is compound sensitive and uses toxic fluorine gas.
The RCD responds to oxidizable species and not selectively to S. The 0-SCD is
by far the best option since it is selective for S, to which it responds quantita-
tively (irrespective of the compound) and is far less susceptible to quenching or
interference and hence should be satisfactory with modified SFC. Its use with
open tubular capillary, packed capillary and packed microbore columns has been
demonstrated with C02 as mobile phase [42]. Earlier thoughts that large quanti-
ties of C02 would quench the chemiluminescence and make packed column un-

320 Chapter 1I
usable have proved unfounded. On-column detection limits are very similar for
capillary and packed columns. The 0-SCD could be used with 5% MeOH modi-
fied C 0 2 [42c].
11.2.4.3.4 Other element selective detectors. Other selective detectors that have
been used for SFC include inductively coupled plasma emission detectors and
electron capture detectors.
11.2.5 Restrictors
Some form of post-column-restriction is required to maintain the pressure of the

mobile phase above its critical pressure throughout the whole separation process
(i.e. in the column). The performance characteristics of the restrictor, which may
be of the non-variable, fixed or variable type, are extremely important especially
in capillary SFC systems. Variable restrictors can be operated manually or pro-
grammed during the course of the SFC analysis. For high pressure liquid phase
detectors (e.g. W D ) the design of the restrictor is not as important provided it
maintains a stable pulse-free flow and does not plug easily. For gas phase detec-
tors the design requirements are more demanding because of the need to quanti-
tatively transfer all the analytes (especially non-volatile and low solubility com-
pounds) to the detector. Ideally the length of the pressure reduction zone be-
tween the column and the detector should be short enough that the fluid main-
tains its density (high solvating power) right up to the entrance to the detector.
1I . 2.5.I Fixed restrictors
Simple fixed restrictors, made out of fused silica or metal capillary tubing, are
used in most SFC systems and particularly in those designed for use with capil-
lary or small diameter packed columns. The major drawback with these restric-
tors is that the mass flow of a fluid through a fixed restrictor increases rapidly
with pressure/density programming, resulting in higher linear mobile phase ve-
locities and lower column efficiencies.
The simplest restrictor, known as a linear restrictor, is made from a short
length (3-15 cm) of fused silica capillary tubing (5-25pm). The geometry of
these restrictors is far from ideal and the long pressure reduction zone results in
poor transfer of analytes with low solubility. The relatively large IDS, however,
make these restrictors less prone to plugging when used with high flow rate sys-
tems (packed columns and SFE). The tapered and integral restrictors represent
attempts to approach the ideal geometry; comparative tests indicate that these
designs are more efficient in transporting the higher molecular mass and less
volatile compounds to the detector [43]. The smaller orifice (1-2 pm) of the ta-
pered and integral restrictors makes them more prone to plugging than either the
linear or frit restrictor. The frit restrictor contains a porous ceramic frit which is
not easily plugged because of its multichannel pore structure. Despite some evi-
dence suggesting poorer transmission of non-volatile solutes, this robust device
is now widely used in many SFC systems.
Metal restrictors (platinum, platinum-iridium and stainless steel) are prepared
by crimping the end of the tubing until the desired flow rate is obtained. Their
performance is comparable to the tapered and integral restrictors although they
work best with moderate to high flow rates (0.25 ml/min upwards). In the
authors experience they are less likely to plug than tapered or integral restrictors
due to the excellent heat-conducting properties of the metal and because manual
crimping often produces more than one orifice. The main disadvantages are a
tendency for the restrictor to open up over a period of time and the irreproduci-
ble nature of the crimping process.
As the supercritical fluid expands from the supercritical to the gaseous state,
severe cooling occurs as a result of the Joule-Thompson effect resulting in the
build-up of solid carbon dioxide or other supercritical fluid at the restrictor tip.
Some heat input is therefore required to prevent the fluid from freezing. In capil-
lary SFC, where column temperatures are often above 100C, the mobile phase
will provide some of the necessary heat which can be supplemented by heaters in
the detector. Packed column SFC is frequently carried out at much lower tem-
peratures (e.g. 40C) and higher flow rates and so must rely entirely on the de-
tector for heating the restrictor.
11.2.5.2 Variable restrictors
The first commercial SFC instrument for packed columns was equipped with a
manually adjustable needle valve (back pressure regulator) which allowed inde-
pendent control over the mobile phase linear velocity but only under isobaric
conditions [44]. An electronically controlled version of the back pressure regula-
tor, in which the needle valve is rapidly pulsed under the control of a solenoid
valve, was described by Saito et al. [45].This variable restrictor, now commer-
cially available, allows flow and modifier gradient programming for isobaric
packed column SFC.
In 1991, Morrissey et al. described a computer-controlled pressure valve unit
which was capable of simultaneously generating both pressure and solvent
modifier gradients for packed column SFC [46]. This design was developed fur-
ther into a commercial system with microprocessor-control gradient program-
ming facilities for mobile phase pressure (density), composition and flow. The
only non-progammable parameter is temperature which is of only minor incon-
venience since most packed column SFC is carried out under isothermal condi-
tions.

322 Chapter I I
Attempts to bring the same sort of control to mobile phase parameters in

capillary SFC have been less successful. The main objective in capillary SFC is
to programme a decrease in the linear velocity of the mobile phase as the pres-
sure/density is increased in order to maintain high column efficiencies. The
cheaper simpler designs have a limited range over which they can alter the linear
velocity while restrictors with a wider and more practical dynamic range are
both expensive and complex.
Recently, a complete SFC system with capabilities for independent pro-
gramming of pressure (density), composition, flow and temperature was
launched. Significantly this system can be configured for either packed column
or capillary SFC modes and finally brings together all the operational parameters
for complete SFC optimization.
11.3 APPLICATIONS
At least 95% of SFC applications in petroleum analysis occur in one of three

areas: simulated distillation, hydrocarbon group type separations and the analysis
of polycyclic aromatic hydrocarbons (PAH). Both the simulated distillation and
hydrocarbon-type analyses use predominantly C 02 as the mobile phase because
they require the use of the FID for quantification. PAHs can be detected and
quantified using spectroscopic detectors, e.g. W D and FLD, and therefore a
wider range of fluids have been used to analyse these compounds. The analysis
of chemical additives which are blended into fuels and lubricants as performance
enhancers has suprisingly received very little attention.
11.3.1 Simulated distillation
Distillation is the primary separation process in the petroleum industry. Knowl-

edge of the boiling point distributions of crude oils and refined products is es-
sential for the operation and control of refinery plant and processes, for product
specification and for quality assurance. Distillation data are obtained using true
distillation methods (e.g. ASTM D86 and D1160) or by analytical techniques
which mimic the distillation process. By far the most widely used technique for
simulated distillation is GC-FID (e.g. ASTM D2887). Conventional distillation
yields true boiling point data and meaningful product fractions, which can be
fbrther characterized in terms of their physicochemical properties, but is expen-
sive and time consuming to perform. Simulated distillation methods are inex-
pensive, rapid and amenable to on-line monitoring but provide no additional data
on product quality.
The upper boiling point limit for ASTM D2887 is 538C which corresponds
to about 85% of a light crude (e.g. West Texas Intermediate or North Sea Brent),
70% of a medium crude (e.g. Arabian Medium) and only 50% of a heavy oil
(e.g. Boscan). Using high temperature GC, this upper limit can be raised above
800C [47], enabling approximately 95% of a light crude oil to be characterized.
To achieve these high boiling points, GC column temperatures up to 430C must
be used and this has naturally raised doubts regarding the thermal integrity of
samples during the analysis [48]. Schwartz et al. [48] presented evidence, using
thermo-gravimetric analysis, that an Arabian Heavy atmospheric residue started
to decompose at around 370C. In practice, laboratories performing routine high
temperature GC simulated distillation, including the authors, have not reported
problems with Arabian Heavy or any other crudes [49].
Even if current high temperature GC methods do not compromise the thermal
integrity of petroleum samples, it is questionable as to how much further GC can
go. The attraction of performing simulated distillation by SFC is that high tem-
peratures are not needed and the mechanism for extending the upper limit de-
pends on sample solubility not volatility. The concept of a simulated distillation
method based on the solubility of sample components in supercritical C 0 2 may
seem implausible at first. Polar high molecular weight components such as as-
phaltenes are not expected to be soluble in C 0 2 as a recent study comparing
Column: 51-17X 50pm SB--Phenyl-5

Program: Linear Density
Mobile Phase: CO, @ 140%
Detector: FID @ 439C
Time Split Injection
Injector @ 11 5%
t L 1
0 16 32 40 60
TIME IN MINUTES
Fig. 11.5. SFC chromatogram of polyethelene standard 750. Conditions: C02 at 140C; linear
density program; FID at 430C;timed split injection with injector at 115C.

324 Chapter I I
13 18 21 28 I1 18 43 48 53
RETENTION TIME (MINUTES1
Fig. 1 I .6. Calibration curve of retention time versus boiling point for polyethylene standard 750.
distillation fractions obtained by molecular distillation with supercritical fluid

extracts appears to confirm [50]. A tripartite relationship between volatility,
molecular weight and solubility is, however, generally recognized and has, for
example been utilized by Boduszynski as part of an overall scheme to separate
and characterize crude oils [ 5 11.
SFC and GC simulated distillation methods are basically the same. Normal
paraffins (e.g. a polyethylene standard like Polywax 750) are used to generate a
retention time versus boiling point calibration for the chromatographic column.
Fig. 11.5 illustrates the SFC chromatogram of Polywax 750 and its associated
SIMDIS calibration plot using a capillary column. The oil sample, usually dis-
solved in carbon disulphide, is analysed under identical conditions to the stan-
dard with all data processing and reporting handled by specialized software
packages.
Either short capillary columns coated with non-polysiloxane phases [52] or
microbore columns (1 and 2.1 mm ID) packed with C18 bonded silica can be
used with equal success. Figure 11.6 shows the separation of Polywax 750 on a
short C18 silica column which compares favourably with the same analysis
achieved on a capillary column (Fig. 1 1S). Figure 1 1.6(b) illustrates the analysis
of a crude oil sample on the same microbore column. Crude oils are one of the
more difficult samples to analyse by either GC or SFC simulated distillation be-
cause of the wide boiling range. The analysis times for packed column SFC are
less than those for capillary SFC and can even be marginally shorter than GC-
TABLE 1 1 . 1
COMPARISON OF GC- AND SFC-SIMDIS DATA FOR PACS (TAKEN FROM REF. 37)
Compound True boiling point Deviation from TBP Deviation from TBP
(C) (C) for GC-SIMIDS (C) for SFC-SIMDIS
Naphthalene 218 0 24
Fluorene 298 -1 I 20
Phenanthrene 338 -1 9 4
Pyrene 393 -1 1 0
Chrysene 447 -3 8 -14
Benzofluoranthene 480 -45 -1 1
Benzo[e]pyrene 493 -48 -10
Picene 519 -50 4
SIh4DIS methods [60]. Retention time repeatability is, of course, critical for
simulated distillation. The capillary column holds a slight advantage in this re-
spect because of the long-term detrimental effect that pressure ramping is ex-
pected to have on the packing structure of the stationary phase in the microbore
column. Bartle and co-workers [53] have reported that the use of an n-
octylpolysiloxane stationary phase in capillary columns leads to better corre-
spondence between the retention behaviour of PAHs and n-alkanes of similar
boiling point (Table 11.1). Since the distillation behaviour of PAHs does not
always relate directly to their boiling point, we should be careful not to conclude
that the SFC SIMDIS data approximates more closely to true distillation behav-
iour.
Many of the early problems with SFC simulated distillation concerned sample
injection. In particular injection of polywax standards was often more trouble-
some than the real samples because of precipitation of the higher molecular
weight waxes in the injection valve and subsequent carryover effects. Dissolving
samples in high boiling solvents (decane, xylene, chlorobenzene) and the use of
heated injectors has largely removed this problem. Sample discrimination using
direct injection techniques (e.g. timed split) is certainly no worse than with high
temperature GC.
Newer SFC instrumentation is capable of programming beyond 400 atm (Fig.
1 1.6(a)) which, in conjunction with higher column temperatures, should extend
the upper boiling point limit even further. At present, the boiling range covered
by routine SFC is the same as that attainable by routine high temperature GC.
The addition of carbon disulphide ( 5 mol%) to COz eluted heavy hydrocarbon
wax material up to C150 (Fig. 11.7) [54], which is a little higher than that re-
ported for high temperature GC [57]. Note, however, that the SFC conditions
were severe and the sample had to be injected onto the packed microbore col-

3 26 Chapter I I
-
0 10
Time (min)
20 30
*
Fig. 11.7. SFE-SFC-FID characterization of a heavy hydrocarbon wax using carbon disulphide-
modified carbon dioxide. Peak 1 = chloroform. 140C, 100 x 1 mm ID experimental C 18 modified
silica column; 180 bar to 507 bar at 15 bar/min.
umn by placing it in a small SFE vessel and extracting at 465 atm and 55OC for
45 min.
Schwartz and co-workers were the first to demonstrate conclusively the valid-
ity of the SFC simulated distillation method [48]. SFC-SIMDIS data were com-
pared not only with true distillation but also with GC-SIMDIS and vacuum
thermogravimetric analysis (VTGA). For samples boiling in the range 260-
540C it was possible to demonstrate excellent correlation with actual distilla-
tion data.
Wax properties such as penetration and solidifying temperature depend on the
molecular weight range and content of low molecular weight material while
tensile strength is influenced by the amount and molecular weight distribution of
is0 and cyclo alkanes. Mellor et al. [55], using capillary SFC, analysed a number
of industrial paraffins and waxes and reported that SFC was suitable for finger-
printing high molecular weight paraffins and waxes. Thompson and Rynaski
1561 compared SFC and high temperature GC simulated distillation for the
analysis of wax samples isolated from underground crude oil storage caverns.
Average differences between the two types of analysis were less than 6C. GC
sample injection required both the sample and the syringe to be heated to 60C;
manual on-column injections had to be made as quickly as possible in order to
prevent a significant drop off in the FID response factors for the higher alkanes.
SFC injection was performed by the timed split method using a heated (1 15OC)
injection valve; lower injector temperatures resulted in poor recovery of the
Supercritical fluidchromatography 3 27
higher molecular weight alkanes. The close agreement between the GC and SFC
data implied little or no thermal decomposition in the former and no sample dis-
crimination problems in the latter.
The distillation characteristics of middle distillate feedstocks to an ethylene
cracker can be monitored using a process supercritical fluid chromatograph. En-
vironmental and economic benefits are claimed by using the SIMDIS data, in
conjunction with hydrocarbon group analysis by GC, to optimize the process
WI.
For certain products such as road bitumens and asphalts, where information
on the distillation characteristics is of little practical value, it more meaningful to
process the SIMDIS data in terms of carbon number distribution. Barbour and
Branthaver [58] used this approach to analyse non-polar asphalt subfractions as
part of the US Strategic Highway Research Program ( S H R P ) . A general model
for asphalt structure is that a non-polar predominantly hydrocarbon solvent
phase acts as a dispersion medium for the more polar aromatic species which
are involved in weak associations (colloids/micelles). The dispersing power of
this solvent phase will be a function of its compositiodconcentration and will
influence some of the important physical properties of the bitumedasphalt.
Since petroleum heavy ends contain significant amounts of material (up to 50%)
which is not soluble in COZY these workers isolated the non-polar component of
the asphalt (the solvent phase) via three different routes: the neutral fraction
from ion exchange chromatography; the low molecular weight fraction from size
exclusion chromatography (SEC); and the maltenes fraction by deasphalting
with iso-octane. SFC-FID analysis was carried out on a short microbore column
(100 x 1 mm ID, polysiloxane stationary phase) using a combined pressure
(135-375 atm) and temperature (125-1 5OOC) gradient. Column degradation was
observed at higher temperatures and pressures. Under these conditions, a carbon
number range from 18-1 10 (relative to a polywax standard) could be studied.
For each asphalt studied, carbon number distributions were similar for all three
non-polar fractions so that any of these fractions might be considered represen-
tative of the solvent phase. For most of the samples it was possible to differen-
tiate between sol-type and gel-type asphalts.
For production purposes, crude oil is usually characterized in terms of
pseudo-components defined in terms of carbon number. Carbon number distri-
butions of 26 stock tank oils were measured using GC and SFC simulated distil-
lation [59]. Both methods were claimed to give the same carbon number distri-
butions within normal experimental scatter although it was reported that for most
crudes the SFC analysis eluted a significantly larger fraction of the oil than did
GC. For light crudes, where the SFC method eluted essentially 100% of the hy-
drocarbons, it was possible to dispense with an internal standard without any
loss of accuracy in the data.
328 Chapter I I
11.3.2 Hydrocarbon group type analysis
Hydrocarbon group type refers to the separation and quantification of alkanes

(saturates), alkenes (olefins), aromatic hydrocarbons and polar compounds in
fossil fuels. Each of these four main group types may be further differentiated
into subgroups, e.g. normal, branched and cyclo-alkanes or mono-, di-, tri- and
poly-cyclic aromatic ring systems.
Until the advent of SFC, this type of analysis was performed using LC, GC or
mass spectrometry (MS). MS provides the most detailed information but is also
the most expensive option and is not often found in routine laboratories. GC is
restricted to the analysis of lighter distillates with FBPs around 220C. Classical
open column LC and HPLC methods are used extensively throughout the petro-
leum industry for routine analysis of middle distillate and higher boiling sam-
ples. Most LC methods rely on a time-consuming and laborious gravimetric
work-up to provide quantitative data because LC does not have a universal detec-
tor similar to the FID in GC. The main attraction of SFC, therefore, lies in its
ability to use the FID for simple and accurate quantification of separated hydro-
carbon groups.
The composition of petroleum changes with increasing boiling point so that a
separation method developed for a gasoline product may not be suitable for die-
sel fuels. Chromatographic methods therefore tend to be quite specific in their
scope with regards the sample types to which they can be applied. There are.
however, a number of common themes and problems which apply to all hydro-
carbon type separations. For example, all LC and SFC methods rely on adsorp-
tion chromatography to separate the different compound types and hence use the
same technology, i.e. silica or bonded silica polar stationary phases.
The SFC method operates with two fundamental differences: the mobile phase
and the FID. Since the mobile phase must be compatible with the FID, this limits
the choice to C02, N20 or SF6. For obvious practical reasons, C 02 is the most
popular fluid. Experience has shown that low fluid densities give GC-like sepa-
rations (i.e. retention increases with decreasing volatility) which are the antithe-
sis of hydrocarbon type analysis. In order to avoid such band broadening effects
within each hydrocarbon group, SFC separations are usually performed with
moderate pressures at or near the critical temperature where fluid densities are
high. It should be noted that it is the density of the fluid which is important, and
not the temperature, implying a solubility and not a volatility phenomenon. An-
dersson has reported that the C02 fluid density should be 0.81 g/ml or higher for
hydrocarbon type separations [61]. Lee and co-workers found that retention was
only slightly influenced by density in the range 0.88-0.93 g/ml [62]. In practice
this means the solvent power of CO, can only be reduced by adding a weaker
FID-compatible fluid such as SFs.
Supercritical jluid chromatography 329
For aromatic hydrocarbon type separations, the influence of alkylation on

the aromatic rings should be minimal. Retention behaviour under GC-like con-
ditions (retention times increase with increasing alkylation) is different to that
under LC-like conditions (retention times decrease with increasing alkylation).
For the majority of hydrocarbon type separations, LC-like conditions are pre-
ferred because they yield compact peaks with less overlap between the different
hydrocarbon types. However, for some analyses, where speciation of a hydro-
carbon group is required (e.g. measurement of naphthalenes in the di-aromatic
fraction of kerosenes), there may be an advantage in operating with GC-like
conditions.
On a solvent polarity scale for silica stationary phases, CO, at densities above
0.8 g/mI would be ranked slightly more polar than the liquid alkanes (pentane,
hexane, heptane) used as non-polar LC mobile phases. In general, hydrocarbon
type separations by SFC with COz are noticeably poorer than those obtained by
LC with hexane. Consequently, techniques used to improve resolution in LC
have also been tried with SFC. In particular, the most difficult separation be-
tween saturates and olefins requires either a more retentive stationary phase
(silver-modified silica retains olefins by complex formation) or a less polar
mobile phase (perfluorinated hydrocarbons in LC, SF6 in SFC). Both options
have been successfully applied to hydrocarbon type analysis by SFC.
SFC allows the use of mobile phase linear velocities above the optimum with-
out sacrificing too much in terms of resolution. The major limiting factor will be
the amount of gaseous C02 entering the FID. Packed capillaries of around
0.75 mm ID can be operated at optimum flow rates with all the column effluent
transferred to the FID. Smaller micropacked capillaries are therefore best for fast
analyses because flow rates above the optimum can be used without compromis-
ing the FID. Microbore columns (1-2 mm ID) must be operated at non-optimum
flow rates if the whole column effluent is transferred to the FID. An option is to
operate at higher flow rates and only send a portion of the column to the FID, a
strategy which must be adopted for larger (>2 mm ID) columns.
Finally, it is worth noting that the constant response of the FID to all hydro-
carbons is an approximation only. Non-uniform response may occur if the FID is
not operated under optimum conditions or the composition of the sample is un-
usual in the nature and amount of saturate and aromatic compounds present. The
presence of non-hydrocarbons, especially sulphur compounds, may exert undue
influence on the quantification which is almost always achieved by normaliza-
tion of the whole chromatogram. Some workers have reported variable FID re-
sponses for saturates and aromatics [63] and for the various aromatic types. As
with all forms of chromatography, problems with quantification can often be
traced to the injector and/or the detector which should be optimized for the type
of sample analysed.

330 Chapter I I
I I . 3.2.1 Gasolines
The fluorescent indicator adsorption (FIA) method (ASTM D 13 19/IP 156) is still
used routinely throughout the oil industry for the determination of saturates,
olefins and aromatics in gasolines and kerosenes. Norris and Rawdon [65] were
the first to suggest that this open column LC method could be replaced with an
SFC-FID equivalent. Using a silica column connected in series with a silver ni-
trate (20%) coated silica column, they demonstrated the separation of a gasoline,
kerosene and diesel fuel into saturates, olefins and aromatics using C 0 2 (35"C,
210 atm) as the mobile phase. The SFC method was quicker (analysis time ca.
5 min), applicable to coloured samples and full range gasolines (cf.. ASTM
Saturates Aromatics
r Temperature ( "C)
50 150
3
20 30 10 40 50
Time (min)
Fig. 1 1.8. 25 x 1 mm ID column packed with 5 p m silica SF6 at 350 atm and 50C.
Supercritical fluid chromatography 33 1
D13 19 where gasolines must be depentanized and coloured contaminants re-

moved by distillation) and showed good repeatability. Results were reported as
% mass by the SFC method compared with % vol. by FIA. The method was
submitted to ASTM for round robin testing but participating laboratories had
difficulty in reproducing the separation; this was thought to be due to instability
problems with the silver nitrate-modified silica column.
Schwartz and Brownlee [66] replaced the dual column system with a single
microbore column (250 X 1 mm ID) packed with silica and switched to the less
polar SF6 as the mobile phase. Although the separation between saturates and
Olefins
+
Saturates Aromatics
Y
1 Baddlush
w 10 20 io 20
lime (rnin) Time (min)
Fig. 11.9. (a) 350 atm 10% C02/SF6,50C, 5 pm silica. (b) 350 atm C o 2 at 40C.

332 Chapter I 1
Aromatics
ins
~ '\ L
4 Backflush
olefins was not as good as that achieved with the C 0 2 and the silver-modified
column [65], the results obtained correlated reasonably well with FIA data. A
pressure and temperature ramp (2.30-340 atm, 50-150C) had to be used after
elution of the saturates and olefins in order to elute the aromatics as a compact
band of peaks. Campbell [67] using identical separation conditions, except for a
larger surface area silica, achieved a much superior separation between saturates
and olefins (see Fig. 11.8). Campbell [67] also reported the use of two different
separation columns which effected a partial but different hydrocarbon type sepa-
ration on each column. Using 10% C 0 2 in SF6 as the mobile phase and a silica
column, the unresolved saturates + olefins elute first and the aromatics are back-
flushed off (Fig. 1 1.9(a)). With C 0 2 as the mobile phase and a silver-loaded sul-
phonic acid silica column, the saturates only are eluted first and unresolved ole-
fins + aromatics are backflushed off the column (Fig. 11.9(b)). Peak areas were
normalized and the olefin content determined by difference. In practice, this
system worked well although it is not suitable for low olefinic samples because
this hydrocarbon group is determined by difference.
This idea was then developed further into a multidimensional SFC system
enabling saturates, olefins and aromatics to be quantified in a single chroma-
tographic run of around 50 min (Fig. 11.10). The sample was injected onto the
silica column where the saturates + olefins were separated from the aromatics.
The aromatic fraction was isolated by valve switching while the saturates and
olefins were separated on the silver-loaded column. After backflushing the ole-
fins off the silver-loaded column, the aromatics were switched back into the
system and eluted [68]. The multidimensional method is applicable to higher
boiling samples including diesels but suffers from two practical disadvantages:
relatively complex and expensive instrumentation and the use of SF6 which pro-
duces the toxic and corrosive HF in the FID. Moreover, the widespread use of
oxygenates and polar additives in fuel will have a detrimental effect on the sta-
bility of the silver-loaded columns.
Anderson et al. [69] reported a similar multidimensional SFC method which
used a different elution order (aromatics were eluted first while the saturates and
olefins were parked on the silver-modified cation exchange column) for the
hydrocarbon types. This system uses COz as the mobile phase and therefore
overcomes objections regarding the use of SF6. By using micropacked capillar-
FID
Saturates
I I I 1
4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0
Mlnutes
Fig. 1 1 . 1 1 Dual SFC chromatograms of standard gasoline RR2.

334 Chapter 11
ies, the analysis times are drastically reduced to around 6 min and the aromatic
fraction is separated into mono- and di-aromatics.
Schulz and Genowitz [70] proposed a much simplified SFC-FID procedure
using a single silica column, COz mobile phase and dual UVD/FID detection.
Samples are injected neat and analysed at constant density using forward elution
only. On this system, the aromatics are completely resolved whereas the satu-
rates and olefins are only partially resolved from each other (see Fig. 1 1 . 1 1). The
UVD is operated at 190 nm where it functions as a quantitative olefin-selective
detector by virtue of its relatively uniform response to olefins. The UVD is cali-
brated against a standard olefin or a sample of known olefin content and the FID
acts as a general mass-sensitive detector for all three hydrocarbon types. Olefin
contents down to 0.02% can be measured and the results correlate closely with
FIA data. Pressure programming can be utilized to sharpen up the aromatic peaks
and so improve detection limits [71].
i i . 3 . 4 . 2Kerosenes and naphthas
SFC methods developed for gasoline analysis are usually suitable for kerosenes
and naphthas. Aviation kerosenes must typically meet specifications on olefin,
total aromatic and naphthalene content. Olefins and total aromatics are deter-
mined by FIA while naphthalenes are measured by a spectrophotometric method
(ASTM D1840). A modified version of ASTM D5 186 (aromatic content in die-
sel fuel by SFC) has been proposed for the determination of aromatic types in
aviation fuel. An additional performance check, using tetralin and 2-
methylnaphthalene, has been included in the kerosene method to ensure suffi-
cient resolution between mono- and di-aromatics. Since the di-aromatic hydro-
carbon group contains several different compound types, but predominantly
naphthalenes and biphenyls, it may not be possible to correlate the SFC results
with those of ASTM D1840.
Experimental design optimization studies for the hydrocarbon type analysis of
aviation fuels have indicated that the optimum resolution for saturates and mono-
aromatics has to be compromised in order to attain the resolution demanded for
the mono-/di-aromatic separation [72]. Fortunately SFC conditions (55OC and
150 atm) were identified which met the resolution requirements and produced a
good separation of all three hydrocarbon types in a kerosene (Fig. 1 1.12).
Di Sanzo and Yoder [63] undertook a wide ranging validation procedure of
their SFC-FID method for jet fuels. The conditions used by these workers (silica
column, C 0 2 at 30C and 115 atm) favoured the separation between saturates
and mono-aromatics at the expense of the aromatic type separation, in agreement
with the optimization studies of Fraile and Sanchez [72]. Jet fuel blends of
known saturate/aromatic content were prepared fiom pre-separated fractions and
analysed by SFC. A small positive bias (1-2% RSD) in the total aromatic con-
I- 1-
AROMATICS
l
i
Fig. 11.12. Separation of aviation fuel under optimized conditions.
tent by SFC was observed, possibly indicating slight differences in FID response
to saturates and aromatics. The quantification of naphthalenes was evaluated by
spiking known amounts of a dimethylnaphthalene mixture into jet fuel saturates
before analysing by SFC. The agreement between actual and measured values
was excellent.
LC methods use amino-bonded or polar amino/cyano-bonded stationary
phases to minimize the effects of alkylation on the retention characteristics of
aromatic compounds. This aspect of stationary phase selectivity has not been
explored to any great extent in SFC. Figure 11.13 illustrates how these charge
transfer type columns. when operated under high density(1iquid-like) conditions,
reduce the influence of aromatic alkylation to give more compact peaks which
are easier to integrate and give lower detection limits [73].
Olefin contents are very low in aviation fuels and are difficult to quantify ac-
curately. The dual UVD/FID method of Schulz and Genowitz [70] may have
sufficient sensitivity as Fig. 11.14 shows.

336 Chapter 1 I
.o
2.5 5.0 7.5 10.0

I
12,s
I I * 1 1
15.0
e 8 % * I "
17.5
# ' 1
.o
Il.. I.l""I..l
Fig. 11.13. Partisil PAC column at 240 bar/50C or 125 bad40"C; 250 X 2 mm ID, COz FID,
aviation fuel.
11.3.2.3Diesel fuels
The first standard method for hydrocarbon type analysis of diesel fuels, based on
HPLC with refractive index detection. was published in 1990 [74]. At the time
One-Ring Aromatics
4.0 5.0 6.0 7.0 8.0 9.0
Mlnules
i
10.0 11.0 12.0 13.0 14.0
Fig. I 1.14. UV and FID chromatograms of a jet fuel (jet fuel 1 190-208 ASTM).
the HPLC method was undergoing development and validation in Europe, SFC-
FID methods were also under study in the US. In 1992, ASTM D5186 was pub-
lished as the first standard SFC method. Both the HPLC and the SFC methods
were developed to meet an urgent analytical requirement, viz the determination
of aromatic content of diesel fuel. Aromatic compounds in diesel fuel are gen-
erally considered responsible for the emissions from diesel engines. Legislation
to control and specify the maximum aromatic content allowable in diesel fuel is
already in force in come countries. Olefins may cause coating of injection noz-
zles and valves leading to impaired combustion and more toxic emissions.
Many laboratories use a simple column and follow a method similar to or
identical to that prescribed in ASTM D5 186. Lee et al. [62] reported that a single
silica column with C02 as the mobile phase gave repeatable results which corre-
lated with FIA data. The aromatics elute over a relatively broad range of reten-
tion times which can make integration difficult particularly for low aromatic die-
sels. Poor precision can often be traced to difficulties in the integration step
when the hydrocarbon group is spread out along the baseline. Di Sanzo and Yo-
der [63], using a similar SFC set-up, also reported problems determining accu-
rate aromatic contents when diesel fuels contained less than 10% mass aromat-
ics; errors were attributed to incomplete separation of saturates and aromatics.
These workers used specially prepared blends of diesel saturates and aromatics
to confirm the excellent accuracy and precision of the SFC method over the
range 10-50% mass aromatic content. Chen and colleagues [75] compared SFC

338 Chapter 1I
Analysis Name : [RSXI 1 DIESELS, 3. 1.

OIESELf . Amount : 1.000.
~ ~ " " 2 ". 0 ' i " 3.0

" " ' i 4". 0 ' 5.0 .o
Tim hinutea)
Instrument : Method : SFC

Channel T i t l e : C a l i b r a t i o n : NONAHE
Lims ID : o Run Sequence : SFC
Acquired on 13-FIAR-1988 a t 16:31
Reported on 19fEB-igW it 17:38
Fig. 1 1.15. SFC-FID analysis of diesel fuels (100 mm X 4.6 mm ID 3 p m SiO, column) with col-
umn backflush to elute the aromatics.
aromatic ring type distribution with D2425 MS data obtained on the aromatic
fractions isolated by LC. The SFC results showed good agreement with the gra-
vimetric aromatic contents and with mono- and di-aromatic data from the MS
analysis; the relationship with tri-aromatic contents was fair.
It may be preferable to analyse low aromatic fuels using pressure program-
ming to compress the aromatics peak [71]; the effect of the increased C 0 2 flow
on FID response should, however, be checked out. Alternatively aromatics can
be backflushed off the column as a sharp band for easy integration and quantifi-
cation (Fig. 1 1.15). Large diameter columns (4-5 mm ID) operated close to or
above their optimum linear velocities yield rapid efficient separations (Fig.
1 1.16) and are a practical alternative to the more extensively used microbore
columns. Dual UV/FID detection can be employed because only a small portion
of the effluent is sent to the FID; the UV detector is a useful adjunct as it pro-
vides a fingerprint of the aromatic distribution and helps to define the satu-
rate/aromatic cut point in low aromatic content samples. The analysis of sulphur
compound types in diesel fiiels using a silica/C02 separation with dual
SCDKJVD has been described [42c].
Supercritical jluid chromatography 339
T i lmin)
Fig. 11.16. Aromatic content in diesel fuel. SFC with dual detection. Conditions: column, 4 x
250 mm silica ( 5 pm); mobile phase, C02; flow rate, 5 ml/min; column temperature, 30C; outlet
pressure, 150 bar.
Fraile and Sanchez [72], using experimental design optimization, have defined
the conditions of pressure and temperature required to meet the D5 186 resolu-
tion requirement for the separation between docosane and toluene. Maximum
resolution was achieved at 190 atm and 32C although a resolution >4 was at-
tained if pressures >120 atm and temperatures <40"C were used. These limits
show good agreement with empirically derived conditions for diesel analysis. A
recent study has noted that silica stationary phases with surface areas >350 m2/g
are more likely to meet the D5186 resolution requirement than low surface area
silicas [75].
Multidimensional approaches to diesel fuel hydrocarbon type speciation are
also documented. Fuhr and co-workers [64] reported the use of a dual column
system (silica and amino-bonded silica stationary phases) for the analysis of
aromatic types by SFC-FID (C02/4000 pd35"C). Diluted fuel samples were in-
jected onto the silica column to effect the saturates/aromatics separation; after
elution of the saturates to the FID, the aromatics were diverted onto the amino-
bonded column to give an improved aromatic-type separation. The validity of the
aromatic-type separation was confirmed by SFC analysis of both model com-
pounds and diesel mono-, di- and poly-aromatic fractions isolated by preparative
LC (Fig. 11.17). Analysis times were roughly three times longer than those on a
single silica column and the additional band broadening of the PAH fraction
could give rise to integration problems. Approximate FID response factors of
1.00, 1.05 and 0.87 were found for the mono-, di- and polyaromatics, respec-
tively; the authors considered these to be sufficiently close to allow aromatic

340 Chapter I I
r I I I I
0 5 10 15 20
lime (min)
Figure 11.17. SFC chromatograms of (a) a whole middle distillate and (b) fraction 1, (c) fraction 2,
and (d) fraction 3 obtained from the whole middle distillate. Conditions are described in the text.
type contents to be determined using area normalization. Repeatabilities for all

aromatic types were around 1% mass which is, not unexpectedly, poorer than
that achieved for total aromatics (0.4% mass) using a single silica column [62].
The SFC results compared favourably with aromatic-type data determined by a
MS matrix method.
Anderson et al. [61,69,76] have described a number of multidimensional SFC-
FID systems using micropacked capillaries and carbon dioxide mobile phase.
The first instrument used two columns (silica and silver-modified cation ex-
change) to separate saturates, olefins and aromatic-types in under 10 min [53].
Separations were performed isobaricaily (300 atm) and isothermally (65C). A
higher column temperature was required to reduce the strength of the olefin/Ag+
interaction and so allow the olefin group to be eluted as a narrow band. To com-
pensate for the higher column temperatures, higher column pressures were re-
quired to maintain densities of >0.82 g/ml. The success of this method is criti-
cally dependent on the availability of well-packed capillary columns which are
stable under conditions of repeated backflushing. The next generation instrument
[76] introduced an additional cyano-bonded silica column to isolate and quantify
polar compounds in diesel. A schematic of the valve and column arrangement is
shown in Fig. 11.18. Higher column temperatures and pressure (75"C, 375 atm,
density 0.81 g/ml) were used to ensure rapid elution of the olefins from the sil-
ver-loaded column. SFC-MS studies were carried out using a micropacked silica
50 pm capillary column, which generated only 50pVmin of gaseous C 0 2 even at
Pca. D.
Poe.c.
w-
J
t
8i
Si
Fig. 1 1.18. Schematic diagram of the column-switching system. (A) Separation on the CN and mSi
columns, polar compounds retained on the CN column, saturates and alkenes transferred to the Ag
column; (B) aromatics transferred from Si column to flame ionization detector; (C) polar com-
pounds backflushed from the CN column; (A) saturates eluted from the Ag column; (D) alkenes
backflushed from the Ag column.

342 Chapter 1I
aU.l
4.w I zw Y
Fig. 11.19. Supercritical fluid chromatogram (FID) of two diesel fuels, CEN 17 and 20, obtained
with the coupled system. Columns: fused silica (25 mm x 0.25 mm ID), packed with Deltabond-
SFC. cyano-bonded, 5pm; fused silica (290 nun X 0.25 nun ID), packed with Superspher Si 60,
4 p m ; fused silica (100 mm X 0.250 mm ID), packed with Nucleosil 5 SA and impregnated in situ
with AgN03. Mobile phase, carbon dioxide at 75C and 375 a m . Peaks: MA, monoaromatics; DA,
diaromatics; TA, triaromatics; P, polar compounds; S, saturated compounds; A, alkenes. Switching
points are indicated by sw.
high column pressures, in forward elution mode only. The saturate fraction con-
tained no aromatics but some alkanedalkenes eluted with the mono-aromatic
fraction. Di Sanzo and Yoder found that SFC-FID gave high aromatic content
values when analysing low aromatic diesel blends [63]. Comparison with IP391
(HPLC with refractive index detection) data shows lower mono-aromatics
(trapped with olefins in the SFC method) and higher di-aromatics (FID response
factor) by SFC; PAH contents are in reasonable agreement although the scatter is
high.
11.4 REFERENCES
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23 T. Greibrokk, B.E. Berg, A.L. Blilie, J. Doehl, A. Farbrot and E. Lundanes, J. Chromatogr.,
394 (1987) 429.
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25 S. Hofinann and T. Greibrokk, J. Microcol. Sep., 1 (1989) 35.
26 D. Tong, K.D. Bartle, R.E. Robinson and P. Altham, J. Chromatogr. Sci., 3 1 (1993) 77.
27 J.C. Kuei, K.E. Markides and M.L. Lee, J. High Resolut. Chromatogr. Chromatogr. Commun.,
10 (1987) 257.
28 J.P. Chervet, M. Ursem, J.P. Salzmann and R.W. Vannoort, J. High Resolut. Chromatogr., 12
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35 K. Sheu, G. Lavigne, H. Sandhu, M. Koenig, G. Moebus and J.F. Johnson, Fuel Sci. Technol.
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Supercritical Fluid Chromatogr Extr), Elsevier, Amsterdam, 1992, pp. 103-127.
37 L.A. Allen, T.E. Glass and H.C. Dom, Anal. Chem., 60 (1988) 390.
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39 J.C. Fjeldsted, R.C. Kong and M.L. Lee, J. Chromatogr., 279 (1983) 449; P.A. David and M.
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E.R. Adlard Fd.), Chromatography in the Petroleum Industry
0 1995 Elsevier Science B.V. All rib&reserved 347
CHAPTER 12
HPLC and column liquid chromatography
A.C. Neal
Esso Research Centre, Milton Hill, Abingdon, Oxfordshire OX13 6AE, UK
12.1 INTRODUCTION
The development of high performance liquid chromatography (HPLC) was

predicted in 1941 by Martin and Synge [l]. In addition to pioneering liquid-
liquid chromatography and the theoretical plate model of chromatography, these
authors predicted that HPLC would be achieved by using very small particles
and a high pressure difference across the column. In fact, the origins of such
columns can be traced back to the work of Tswett in 1903 [2] and their use was
further extended by Kuhn and Lederer in 193 1 [3].
The advance of gas chromatography (GC) in the petroleum industry in the
1950s was such that liquid chromatography was effectively overlooked through-
out that decade. This rapid exploration and application of GC rekindled interest
in liquid chromatography as a complementary technique which could open up
regions of solute polarity, molecular weight and bulk separation alien to GC.
Commercialization of HPLC columns, pumps and detectors during the 1960s and
early 1970s simplified operation of the technique and allowed potential users to
apply it with relative ease. During this time, various terms were used to describe
the new technique: modern liquid chromatography, high pressure liquid
chromatography and high performance liquid chromatography and the latter
is now universally used although it is not easy to define exactly what is meant by
high performance. A comprehensive introduction to HPLC can be found in
Snyder and Kirkland [4] although this excellent book is now almost 20 years old.
More recently, Parris [5] and Gilbert [6] have written good general reviews on
the subject and the reader is referred to reference [4] for detailed and fundamen-
tal information and the latter two books for general theory and instrumental de-
scriptions.
348 Chapter 12
The applications of HPLC in petroleum analysis has itself been reviewed by

Amos in 1979 [7]. At that time, the number of specific applications in the indus-
try were few and the major uses were in hydrocarbon type analysis and the de-
termination of hindered phenol antioxidants in fuels, most notably aviation tur-
bine fuel. By comparison with paper chromatography and thin-layer chromatog-
raphy, which had independently competed with HPLC during the earlier years of
development, Amos concluded that the choice between PC, TLC and HPLC is
now fairly clear cut and that HPLC should be used for all routine high-speed
quantitative analysis.
Since that review was written, HPLC pump, column and detector designs have
advanced and expanded markedly, such that a far wider field of applications now
exists. HPLC has also diversified into aqueous/ionic systems (ion chromatogra-
phy) and high performance size exclusion chromatography (gel permeation
chromatography), and been hyphenated with spectrometry including induc-
tively coupled plasma emission spectroscopy (ICPES), nuclear magnetic reso-
nance spectroscopy (NMR), Fourier transform infra-red spectroscopy (FTIR)
and most importantly mass spectrometry (MS). The numerous attempts to inter-
face HPLC with MS have resulted in a variety of LC-MS systems with each in-
terface type having its own specific limitations and applications.
12.2 APPARATUS
A typical HPLC system (Fig. 12.1) is still composed of a pump, sample injec-
tor, column, detector and data recorder much as described by Amos [7]. How-
ever, considerable improvement and development of each component has taken
place. Advances in columns and detectors have resulted in a wider range of sepa-
rations and detection strategies being available.
12.2.1 Solvent reservoirs
Solvent reservoirs consist of purpose built glass bottles with a helium inlet
and filter (for degassing) and a solvent outlet composed of a fritted particulate
filter and PTFE outlet tube. Some systems even allow for a slight helium over-
pressurization of the reservoir to assist pump priming and prevent cavitation in
the solvent inlet tubing.
12.2.2 Pumps
Pumps have progressed from single isocratic systems delivering premixed

solvent, to purpose built binary, tertiary and even quaternary mixing systems
HPLC and column liquid chromatography 349
Solvent Reservoirs
Detectors Data System

Fig. 12.1.Elements of an HPLC system.
which premix the solvents to the desired composition and deliver them at the
required flow rate, compensating automatically for pressure and viscosity effects
which may occur during the mixing process.
During the early 1980s, much debate took place over the relative merits of
high pressure mixing (after the pump outlet) and low pressure mixing (at the
pump inlet).
High pressure mixing suffers from a number of drawbacks: chief among these
is the need for more than one pump with the concomitant expense. In addition,
imprecision in the solvent composition may occur if one or more of the solvents
is present as less than 5% of the total.
Low pressure mixing requires only one pump with the solvents proportioned
and mixed before the pump head. Control by microprocessor or computer data
systems allows for almost any shape of gradient (and flow) profile to be deliv-
ered. For these reasons, low pressure mixing, under either of the remote control
systems given above, has come to dominate the market for LC gradient systems
but some caution is still necessary in use. Firstly, for complete mixing, some
systems rely on a fairly large volume mixing chamber on the outlet side of the
pump. In certain applications, such as backflushing, a sharp gradient profile is
desirable and this may be compromised by the hold-up volume of the chamber.
In other words, if it is necessary to make sudden step changes in the gradient, the
step may actually be a slope. Low volume ( 1 0 ~ 1 dynamic
) mixers, such as the
Referencespp. 3 72-3 74
350 Chapter 12
LEE micromixer, are of considerable use in eliminating this problem. Secondly,

efficient degassing of the solvent used to be a prerequisite for accurate mixing in
order to eliminate cavitation in the mixing system and the detector noise and in-
accurate flow rates that could result. In practice, improvement in the design of
the solvent reservoirs and pump head geometry have reduced the occurrence of
these problems, provided that the manufacturers advice is heeded. Gradient elu-
tion may also limit the choice of detector to be used, especially if the detector
depends on changes in a physical property of the mobile phase itself. This issue
is discussed in detail in the section on detectors.
Three main types of pump, reciprocating piston, syringe and diaphragm, were
all applied in the early days but the reciprocating piston pump now dominates
the standard HPLC market where flow rates are at , or above, 1 ml min-*. Sy-
ringe pumps are best suited to lower flow rates and as such find more use in
microbore HPLC where flow rates are typically well below 1 ml min-l.
Reciprocating piston pumps operate by means of a rotating eccentric cam
which drives a piston. The piston draws solvent into a cylinder through the inlet
check valve during the return stroke. During the delivery stroke, solvent is ex-
pelled through the outlet check valve and hence ultimately to the column. These
pumps are relatively inexpensive, simple to maintain, and deliver a constant flow
of solvent over a wide range of flow rates. The piston drive is usually controlled
by solid state pulsing circuits and a stepper motor. This allows for rapid refilling
of the cylinder followed by swift repressurization of the solvent in the pump
head and then a smooth, constant volume delivery until the end of the delivery
stroke. The design of the cam and its eccentricity determine the smoothness of
the flow profile. This is now so well defined that accurate, rapid refilling has
become commonplace and methods for smoothing the profile such as large vol-
ume pulse dampers and dual or triple stage pump heads are largely redundant.
Lower volume dampers may still be used and are often an integral part of the
pump, invisible inside the box.
12.2.3 Sample injectors
Sample injectors are almost exclusively of the six-port valve type, although
on-column syringe injectors were initially used. Injection valves are connected
between the pump and the column and as close to the top of the column as prac-
tically possible. An interchangeable sample loop of discreet volume is connected
to the valve and isolated from the flow of mobile phase. The loop is filled with
sample solution and the valve is then turned manually or electronically so that
the loop is connected into the flowing mobile phase and the sample is thereby
injected onto the column.
Various designs of injectors exist, depending on the volume of sample avail-

able and the physical scale of the HPLC system. Internal microlitre-sized loop
injectors are available for microbore applications, whereas for standard and
preparative work, interchangeable sample loops up to 5 ml in volume can be
used with standard Rheodyne or Valco valves. Loops are filled by a standard
glass syringe with a Luer fitting, with or without a flat-ended needle according to
the design of the sample inlet on the valve.
Remote actuation of injection valves or flow switching valves is usually ac-
complished pneumatically or electrically. Typically the solvent delivery system
or data station allows for timed events, one of which is the activation of the in-
jection valve. In pneumatic actuation, the valve is turned by a supply of high
pressure gas, usually air, in a purpose made pistodcylinder type actuator. The
gas supply is itself delivered to the actuator by a solenoid valve. The solenoid
valve opens or closes under the control of the solvent delivery system or data
station, in order to pressurize or depressurize the actuator and hence operate the
injector.
Electrical actuators tend to be more expensive but are faster than pneumatic
actuators. They use a synchronous, high torque electric motor, directly con-
trolled by a relay closure or TTL switch. The valve may be coupled directly to
the motor, minimizing the number of moving parts. Although faster and poten-
tially more reliable than the pneumatic actuator, the extra capital cost is often the
sole factor which mitigates against their use. In our experience, pneumatic actua-
tors seldom give any cause for concern and the extra cost of electrical actuators
is rarely justified.
12.2.4 Columns
Selection of the appropriate column is, of course, entirely dependent on the

particular separation desired. Over the last 15 years, the technology of column
design and manufacture has advanced markedly, as has the range and reliability
of packings available. In the late 1970s, columns were almost exclusively
250 mm long 316 stainless steel with an internal diameter of 4.6 mm. End fit-
tings were of solid 3 16 stainless steel and packings typically amorphous silica or
alumina, or else silica with an octadecyl bonded phase, commonly referred to as
ODS or C18. At that time, intermediate polarity stationary phases were begin-
ning to excite interest but only amino (-NH,) and nitrile ( X N ) phases were
readily available. Since then the range of HPLC applications has broadened
considerably and advances in column chemistry and design have been fundamen-
tal to that progress.
HPLC analysis can be placed in one of four categories largely by virtue of the
column type used.
References pp. 3 72-3 74

352 Chapter I2
(1) Reversed phase: where the phase is of spherical silica with a non-polar
hydrocarbon chemically bonded onto the surface or less commonly sty-
rene-divinylbenzene beads. The mobile phase is polar and is most often a
mixture of methanol and water, acetonitrile and water or tetrahydrofuran
and water. One important variant of reversed-phase HPLC is reversed
phase ion-pair chromatography (RP-IPC) where the analyte is ionizable or
protonizable, and the mobile phase consists of a buffered aqueous mixture
containing a counter ion of opposite charge to the analyte.
(2) Normal phase: where the column is packed with spherical silica or with
silica with a polar phase chemically bonded to it. Typical bonded phases
include amino (-NH,) and nitrile (-CN) already referred to (and which
may also be used in reversed phase mode) and phenyl, nitro or diol. Some
specific phases such as dinitroanilinopropyl are also finding considerable
use. The mobile phase is non-polar, typically heptane with or without the
addition of small amounts of more polar solvents such as methylene chlo-
ride or ethyl acetate.
(3) Ion exchange: consisting of sulphonate or quaternary ammonium func-
tional groups chemically bonded onto silica or styrene/divinylbenzene
polymer particles. Weak cations or anions can be separated without the
use of buffer solutions as mobile phase, whereas strong cations or anions
will require them.
(4) Size exclusion or gel permeation: where solutes are separated by virtue of
their size in solution. This technique has many petroleum applications for
the determination of the molecular weight of polymeric lubricant addi-
tives but is not considered in detail in this chapter.
The range of columns currently available is, therefore, extremely wide, such
that the separation of hydrocarbons, functional groups, ionic compounds, poly-
mers and even enantiomers can be achieved. Column design has advanced from
conventional columns to include disposable cartridges, radially compressed
columns, metal-free columns made from polyetheretherketone (PEEK), and col-
umns with adjustable end fittings which recompress the packing if voids de-
velop, prolonging column lifetime.
Most stationary phases are also available in microbore columns with internal
diameters of 1-2 mm, which offer the advantages of reduced mobile phase con-
sumption and greater mass sensitivity. By contrast, preparative scale columns
packed with any of the aforementioned stationary phases (with the exception of
diol) are also available off the shelf. These have internal diameters (i.d.) of 9
or 21 mm and can be used to recover larger quantities of analytes, either for fur-
ther purification or for identification.
A petroleum HPLC laboratory can serve a diverse group of needs including
the analysis of fuels, lubricants, additives, waste water and refinery process
samples. As a consequence one could easily expect to find silica, C18, C8,
-NH,, -CN, ion exchange and size exclusion stationary phases in routine use,
each in one or more of the column designs described above.
12.2.5 Detectors
All detectors, be they for HPLC or any other analytical technique, must be
precise, sensitive and stable. In addition, HPLC detectors should have a large
linear dynamic range, be insensitive to temperature and eluent composition, ex-
hibit low noise and drift, and be simple and easy to maintain. Since the early
days of HPLC, no single detector has been able to fulfil all these criteria as the
flame ionization detector (FID) has done so admirably for GC. Instead, a range
of detectors has evolved, based either on changes in the bulk properties of the
mobile phase, or upon a selective property of the analyte(s). The subject has
been frequently reviewed and descriptions of the main detector types can be
found in any general HPLC text [5,6]. The reader is directed to Scott for a more
detailed and mathematical treatment [81. The treatment here will be restricted to
a brief discussion of those detectors which have found application in the petro-
leum industry. Even with this proviso, the majority of detector types currently
available are still included.
12.2.6 Selective property detectors
12.2.6.I W-visible spectrophotometers

Ultraviolet detectors have been used since the early years of HPLC and re-
main the workhorse detector in the majority of laboratories today. Early exam-
ples were effectively converted spectrophotometers with the only modifications
being associated with inclusion of a flow cell rather than a cuvette holder. These
instruments were therefore based on prism diffraction or grating interferometry,
such that the specific ,A of interest could be selected in order to achieve maxi-
mum solute sensitivity. The quantitation principle is the Beer-Lambert Law
which states that the amount of UV or visible light absorbed will be directly pro-
portional to the solute concentration. Sensitivity and limits of detection will
therefore vary from solute to solute as a function of the individual compounds
extinction coefficient. In extreme cases, where no UV or visible light chromo-
phore is present in the solute, no absorption will take place and such solutes will
not be detectable. This is the chief limitation of UV detectors and it is especially
apparent in petroleum analysis because saturated hydrocarbons have no chromo-
phore. A second major limitation is that mobile phases which themselves absorb

354 Chapter 12
UV light effectively impose a wavelength cutoff on the system. Below this

wavelength the absorption of the mobile phase itself is so strong that solutes
with A, in the same region cannot be detected.
The principle of detection within prism or grating instruments relies on the
transmitted light of the chosen wavelength being cast onto a photomultiplier.
Only one wavelength is observed that is often a compromise between sensitivity
and selectivity.
I2.2.6.2Diode array detectors (DAD)
Diode array detectors (Fig. 12.2) effectively allow a much broader wavelength
range to be acquired simultaneously, such that an entire W spectrum (more
typically 200400 nm) can be captured repeatedly throughout the analysis. These
detectors became commercially available in the early 1980s, have rapidly estab-
lished themselves as reliable and sensitive, and have allowed ever more complex
detection strategies to be employed. Of course the solute still needs a chromo-
phore and the mobile phase W cutoff must be observed, especially in gradient
elution.
The principle of detection within the DAD relies upon an array of photodi-
odes of typically 0.5 nm resolution, such that the transmitted light after the flow
cell is dispersed by a holographic polychromator and directed onto the linear
photodiode array. Thus, by recording the signal output from one photodiode, the
eluent is monitored at a single wavelength and by recording the output from all
Photodiode Array
gg/gg' Ellipsoidal Mirror
ElliosoidalMirror
I # - Deuterium Lamp
Fig. 12.2. Optical system of a diode array detector.

the diodes, an entire spectrum is obtained. The major disadvantage of this system
is that light of all wavelengths is present in the sample cell simultaneously and as
such, fluorescent light may well be present at the wavelengths being monitored.
In practice this is such an infrequent occurrence as to be of little consequence
but it must always be borne in mind, especially if the mobile phase or solutes can
be excited.
The DAD can be used as a simultaneous multiwavelength detector to maxi-
mize sensitivity to each solute in turn, or to record entire spectra in order to ex-
amine peak purity, or to produce three-dimensional maps of wavelength versus
absorbance versus time. All three modes offer the user more accurate quantita-
tion than would be possible with a single wavelength dispersive W spectropho-
tometer. For research use, the ability to record an entire W spectrum of all the
unknowns in a sample can give an early indication of solute identity. Coupled
with retention behaviour, this can yield hypothetical structural information, or
solute functionality, or carbon number, depending on the LC mode employed
(normal or reversed phase).
12.2.6.3 Fluorescence detectors
UV light can interact with some solutes by exciting delocalized electrons into
higher energy states above the normal ground state. When these electrons relax
back to the ground state, the solute will emit most of the absorbed energy as light
at a longer wavelength than that which excited it. In solutes where this decay is
instantaneous or where it ceases immediately upon removal of the incident light,
the solute is said to be fluorescent. It is possible to monitor the emission wave-
length and filter out the excitation wavelength altogether, and this produces very
high sensitivity, some two to three orders of magnitude greater than W absor-
bance and is therefore a highly desirable method of detection. In order to take
advantage of the phenomenon, non-fluorescent compounds may be derivatized
prior to analysis with a reagent to produce a fluorescent derivative. In petroleum
analysis, fluorescence detection is most useful when the solute itself is highly
conjugated and fluoresces naturally, as do many polynuclear aromatic hydrocar-
bons. Fluorescent light emerges from the sample at random angles and most in-
struments monitor the light emitted at right angles to the excitation beam. Some
solvents have the ability to quench fluorescence such that the process is effec-
tively suppressed. In particular very polar or aqueous mobile phases and buff-
ered or ionic eluents are not recommended due to this phenomenon.
12.2.6.4Electrochemical detectors
Compounds which are electrically oxidizable or reducible can be detected
electrochemically.In coulometric detectors the solute is completely electrolysed,
whereas in amperometric detectors, the solute is only partially electrolysed.

356 Chapter 12
Amperometric operation is more suited to flowing systems and is the commonest

mode of electrochemical detection. In amperometric detectors, the solute con-
centration is directly proportional to the diffusion rate of the solute across
the boundary layer to the electrode surface. The electrode current is therefore
dependent not only on the solute concentration but also on its diffusion coeffi-
cient. A detailed treatment of electrochemical detectors can be found in
Scott [8]. Electrochemical detectors rely on the mobile phase being electri-
cally conductive and the most direct method of assuring this is to use buffer so-
lutions.
12.2.6.5Flame ionization detector
The use of the FID in HPLC necessitates the removal of the mobile phase,
chiefly by selective evaporation. Much effort has been expended into making the
FID compatible with HPLC in order to take advantage of its properties of sensi-
tivity, known response and linear dynamic range. A number of mechanical trans-
port systems have been developed originating with James in 1964 [9]. A moving
wire was employed to carry the column effluent through a heated zone where the
mobile phase was evaporated off, and then to another zone heated to a higher
temperature in order to evaporate/pyrolyse the solutes and carry them into the
FID in a stream of nitrogen.
The chief disadvantages of transport detectors all lie with the transport
mechanism itself. The wire, chain or disk has proved to be difficult to coat uni-
formly, different solvents evaporate at different rates, accumulation of remaining
traces of solute give memory effects. These factors all contribute to relatively
poor signal to noise ratios. Since only a small proportion of the solute is evapo-
rated and detected, the sensitivity and large linear range of the F D are not util-
ized. In conclusion the compromises inherent in transport FIDs have meant that
this detector is not widely used and its early promise for HPLC use remains
largely unfulfilled.
12.2.6.6 Mass spectrometers
The interfacing of mass spectrometers to GC instruments (GC-MS) is possible
because both techniques are readily compatible. GC-MS is now one of the most
powerful diagnostic tools available to analytical chemists.
Interfacing HPLC to mass spectrometers (LC-MS) is much more difficult and
has largely hinged on the design of liquid phase separation systems and removal
of eluent until relatively recently. Work on LC-MS began in the late 1960s but it
was not until the work of Homing et al. [lo], Scott et al. [l 13, and Arpino et al.
[ I 21 in the 1970s that LC-MS was effectively achieved.
Overcoming the relative incompatibility of the liquid phase eluent and the
high vacuum required in the source of the MS has proved to be a severe chal-
lenge. In addition, the higher molecular weight, lower volatility and chemical
polarity of many compounds separated by LC make them less easily ionized than
the compounds amenable to GC-MS. Because of this, electron impact ionization
(EI), which is so successful in GC-MS, has proved less so in LC-MS. More rele-
vant ionization techniques such as fast atom bombardment (FAB) and atmos-
pheric pressure chemical ionization (APCI) have been applied in order to sur-
mount this problem. The mere modification of LC to make it more compatible
with existing high vacuum, electron impact MS has not on its own proved suffi-
cient, and the development of more compatible MS ionization and inlet systems
has been necessary for the two techniques to merge successfully. The whole
subject has been well reviewed recently by Niessen and van der Greef [13].
These authors list 26 distinct types of interfaces for LC-MS developed since
1972. The reader should consult reference [131 in respect of thermospray LC-MS
and the particle beam interface, both of which have been successful in petroleum
and coal-based applications.
12,2.6.7Injured and NMR
IR photometers have found little use as HPLC detectors for two main reasons.
Firstly, most solvents used as mobile phases absorb in the most useful regions of
the IR spectrum. Secondly, using absorption wavelengths away from solvent ab-
sorption bands has invariably resulted in less sensitivity and higher background
levels. The exceptions to this have been where the analyte contains a carbonyl
(C=O) group and in size exclusion chromatography of polymers. The former
case is able to take advantage of the high extinction coefficient and hence high
sensitivity of the carbonyl group. The latter application is able to overcome both
low sensitivity and high background by virtue of the relatively high sample con-
centration required by SEC.
Many of the limitations have been overcome or greatly reduced by Fourier
transform infrared (FTIR) instruments. Modern FTIR spectrometers have signal
to noise ratios over 100 times larger than energy dispersive instruments and as a
consequence sensitivity is greatly improved. Their chief disadvantage is that of
high cost and another disadvantage is incompatibility with reversed phase elu-
ents. The combination of water absorption and band broadening due to hydrogen
bonding conspire to reduce sensitivity and to limit the usable part of the IR
spectrum.
Proton nuclear magnetic resonance spectroscopy ('H NMR) has also been
used as an on-line HPLC detector. This technique exploits the odd spin of the
hydrogen nucleus, lH, in order to gain information on the environment of various
hydrogen atoms in the analyte molecules. In this way, the signals due to methyl,
methylene and aromatic protons in various molecular environments can be sepa-
rated and quantified. Once normalized, the proportion of various hydrogen types

358 Chapter 12
can be calculated and the alkyl, aryl and heteroatom substituents present in a
sample elucidated. Proton NMR will be unable to distinguish hydrogen atoms
from the mobile phase from those of the analyte and these will be included erro-
neously in the normalization if present. For this reason, static NMR experiments
or LC-NMR cannot use standard solvents but are required to use perhalogenated
or perdeuterated solvents. This is a severe limitation to on-line LC-NMR since
these solvents are extremely expensive, especially if significant volumes of per-
deuterated solvents such as chloroform-d (where 99.8% of the hydrogen is re-
placed by deuterium) have to be used for the LC separation. Another consider-
able limitation is the high capital and running cost of a modern Fourier transform
NMR spectrometer. Nevertheless, this technique has found application in petro-
leum analysis and is expected to find increasing use.
12.2.7 Bulk property detectors
12.2.7.1 Refvactive index detector

The refractive index detector remains the second most widely used LC detec-
tor after the UV detector. It is universal, detecting all analytes whose refractive
index (RI) differs from that of the mobile phase.
The RI of a substance is a dimensionless constant which typically decreases
with increasing temperature. Three types of RI detector are available and all are
termed differential refractometers, that is they measure the difference in RI be-
tween a sample cell and a reference cell containing mobile phase only. It fol-
lows, therefore, that all refractometers are sensitive to temperature changes and
to changes in eluent composition. Thus , in order to use them for gradient elu-
tion, the reference cell must always contain a mobile phase of identical compo-
sition to that in the sample cell and this is often impossible to achieve. For good
baseline stability, RI detectors are thermostatically controlled, either by a water
bath or by an insulated cabinet.
Deflection or angle of deviation instruments have a split flow cell, with Sam-
ple on one side, reference eluent on the other. Light from the source passes
through this cell to a mirror behind it and is reflected back through the cell to a
photomultiplier. If a solute of different RI enters the sample cell, the light beam
will be deflected. The photomultiplier output is proportional to the magnitude of
the deflection. Deflection RI detectors are simple and have a wide linear dy-
namic range. Instruments manufactured by Waters Associates have typically
been of this design.
Fresnel refractometers pass parallel incident light through a prism onto sam-
ple and reference simultaneously. If the refractive index of the liquid in the
sample cell differs from that in the reference cell, some light from the sample
cell will be diffracted, reducing the intensity of the beam reflected back out of
the sample cell. The difference between the intensities of the sample and refer-
ence beams is measured by a photomultiplier and recorded. The linear concen-
tration range of this detector is less than that of the deflection instrument unless
two separate prisms are used to cover the entire RI range. The optical cleanliness
of the system is also more critical than for the deflection detector. Fresnel refrac-
tometers have been manufactured by Perkin Elmer.
Interference refractometers split the source beam, pass it through sample and
reference cells simultaneously and then recombine it. Any difference in refrac-
tive index between sample and reference cells will manifest itself as a difference
in optical path length when measured by an interferometer. This design is more
sensitive than the previous types and additional sensitivity is possible if a laser is
used as the light source as by Woodruff and Yeung [14,151.
In summary, RI detectors are universal and can be sensitive under carefully
controlled conditions. Their use in gradient elution is still far from straight-
forward and base line drift is to be expected when the mobile phase composition
changes even by relatively small amounts. Despite all these operational draw-
backs, they are still the detector of choice when the solutes have no UV chromo-
phore, especially in isocratic determinations of saturated hydrocarbons.
12.2.7.2Evaporative light scattering detectors
The evaporative light scattering detector (Fig. 12.3), evaporative analyzer or
mass detector was developed and patented in 1966 by Ford and Kennard [16,171.
It was not until 1978, however, and the comprehensive work of Charlesworth
[18] that its usefulness as an HPLC detector was fully realized. The theory of
operation, construction and performance of what is now referred to as the mass
detector can be found in that reference.
In essence, this type of detector consists of a nebulizer, evaporation chamber,
light source, scattering chamber and light trap and a photomultiplier set at 135
to the incident light beam. Column eluate is nebulized with a relatively high flow
of nitrogen or air and the mobile phase evaporated as the solvenugas mixture
passes down the vertically mounted evaporation chamber. At the bottom of the
chamber, all that is left is gas, solvent vapour and finely divided droplets or par-
ticles of analyte. This aerosol passes through the light beam and the photomul-
tiplier detects that portion of the incident light which is scattered by the analyte
(at an angle of 135). At this angle, Charlesworth found the result to be effec-
tively independent of the RI of the analyte.
The true linear working range of this instrument is not extensive, typically 1.5
orders of magnitude in concentration. Above and below this range, the size of
the analyte droplets produced no longer promote the reflection and refraction of
the light. Although this is a drawback, it is a relatively minor one, as the re-

3 60 Chapter 12
Nebuliser aa
n I-
118 I I 1 =Exhaust
Fig. 12.3. Schematic of an evaporative m a s detector.
sponse functions due to Rayleigh and Mie scattering in the non-linear regions are
well described and still make calibration possible.
The chief disadvantage of this detector is the volatility limitations imposed
upon the analyte. The solvent evaporation chamber is, in effect a mild blow-
down apparatus which removes the mobile phase. If the analyte volatility or va-
pour pressure approaches that of the mobile phase it will vaporize and give no
response. In our laboratory, we have found hexadecane (b.p. 256C) to be par-
tially evaporated when the detector is operating at ambient temperature with
hexane (b.p. 68C). It is therefore likely that hydrocarbons below n-CI7will not
give full recovery. Even given this limitation, the detector finds considerable use
for intermediate and low volatility analytes.
12.2.7.3 Dielectric constant detector
With few exceptions, the dielectric constant of a substance increases with its
polarity. As an LC analyte elutes from the column, the dielectric constant of the
eluate will change. The dielectric constant of a non-polar or semi-polar sub-
stance is a function of its refractive index and as such many of the practical con-
siderations concerning RI detectors apply equally to the DCD. A more detailed
treatment may be found in Scott [8].
A typical DCD is a differential, temperature controlled concentric cylindrical

capacitor through which the column eluent flows. The cell electrodes are made
of stainless steel and are connected electrically as one side of a Wein or Schering
Bridge. If the mobile phase is less polar than the analyte, as in normal phase LC,
the dielectric constant of the eluate will increase as the peak elutes. The reverse
is usually the case in Rp-HPLC, and in order to avoid negative peaks in reversed
phase applications, DCDs allow for polarity reversal.
Setting up and balancing DCDs can be a tedious business as each side of the
bridge circuit needs to be balanced in an iterative fashion until the potential dif-
ference across the bridge is zero.
The linear dynamic range of the DCD is heavily influenced by the difference
in the dielectric constants of the mobile phase and the analyte, but has been
quoted as 3.5 X lo4 which is comparable to the RI detector.
12.3 QUANTITATION
In the majority of petroleum applications of HPLC, calibration is by external

standardization and quantitation is by peak area. Where samples are analyzed as
received or after dilution only, this approach is reliable and accurate. Where the
sample is worked up before analysis by liquidliquid or liquidsolid extraction, it
is necessary to determine the extraction efficiency (or recovery) in order to be
certain that a representative extract has been obtained. Where extraction effi-
ciencies are low or where time does not allow the recovery to be determined, an
internal standard or a standard addition method should be employed, provided
the detector response to the solutes is linear in the range of interest.
Peak area is most usually used for quantitation, as this is the most statistically
precise measure of analyte concentration. It does presuppose good resolution
however, and where this is not the case, a range of deconvolution methods or
even peak height measurement may have to be considered.
Contemporary HPLC now has a vast range of competitive quantitation devices
and statisticaVgraphica1 software available. Stand alone benchtop integrators,
microprocessor and PC data stations, local area networks (LANs), laboratory
information management systems (LIMS) and even mainframe chromatography
packages are all available. Selection is a compromise between cost, specification
and, increasingly, compatibility with existing computer hardware. Any of these
devices can take detector output and convert it to a high quality graphical or nu-
merical report, automatically labelled with peak identities according to previ-
ously recorded retention windows.
Caution is necessary, however, as any system will only act according to the
way it is configured by the operator. At each stage of the data systems applica-

362 Chapter 12
tion, the user must be certain that each setting is sound in order to obtain a final
quantitative output of the highest possible integrity.
12.4 APPLICATIONS
The applications of HPLC in petroleum analysis are summarized in Table

12.1. The wide variety of separation mechanisms, column chemistries and de-
tection systems represented by HPLC offers the petroleum chemist a range of
distinct systems. In general these fall into three categories:
(1) the separation and direct quantitation of individual compounds;

(2) separation and characterization of compound classes such as, for example,
saturates, olefins and aromatics in petroleum products;
(3) preparative or semi-preparative fractionation of complex mixtures for de-
termination by other analytical techniques.
Within each category, standard methods exist for particular determinations,

which have been rigorously tested in terms of inter-laboratory precision. Such
standard methods as exist within the Institute of Petroleum handbook, Standard
Methods of Analysis and Testing of Petroleum and Related Products, 1993 [19]
are discussed in the following sections.
12.4.1 Individual compounds
12.4.1.1 Polycyclic aromatic hydrocarbons (PAHs)

These compounds have attracted considerable interest due to their role as
pollutants and, in some cases, their carcinogenic properties. Amos [7] cites some
early W L C applications. Katz and Ogan [20] have used partition and size ex-
clusion columns in series to effect the analysis, and a combination of normal
phase amino and reversed phase C 18 columns has been used to determine PAHs
in crude oil by Grimalt and Albaigks [21]. Further LC-LC methods, chiefly
aimed at benz[a]pyrene, have been employed by Tomkins and Griest [22] and
Fielden and Packham [23]. In the former case, Partisil silica and analytical scale
Vydac 201TP reversed phase columns were used and in the latter case cyclodex-
trin and ODS silica. In both cases, the selectivity and sensitivity of fluorescence
detection was used to determine the PAH directly.
Symons and Crick [24] have determined PAHs in refinery effluent after clean-
up and preconcentration using a Radial-Pak CIS column with 75:25 aceto-
nitrilelwater eluent and UV and fluorescence detection. Recoveries were vari-
TABLE 12.1
APPLICATIONS OF HPLC IN PETROLEUM ANALYSIS
Crude oil
ha-arenes Grimmer et al.
Dibenzothiophene Rebbert et al.
Christensen and White
Phenols MacCrehan and Brown-Thomas
Polynuclear aromatic hydrocarbons (PAHs) Grimalt and Albaiges
Preparation of PAH fractions Ostman and Colmsjo
Saturates/aromatic types Welch and Hoffman
Naphthdgasoline
Aromatic nitrogen compounds Nondek and Chvalovsky
Benz[a]pyrene Tomkins and Griest
Saturateshromatic types Apffel and McNair
Cookson et al.
Munari et al.
Hayes and Anderson
Saturates fractions ASTM D 2002,2003
Saturates/olefindaromatics ASTM D 1319
Aviation fuel
Coumarin IP 374
PAHS Fielden and Packham
Saturateshrornatics IP PM-AT
Saturateshromatic types Welch and Hoffman
Cookson et al.
Hayes and Anderson
Haw et al.
Davies et al.
Saturates/olefindaromatics ASTM D1319
2,4-Dimethyl-6-tertiarybutylphenol Hayes and Hillman
IP 343
DieseUdistillatefuels
Alkyl nitrates Schabron and Fuller
Mono/di/triaromatics IP 391,PM-AY
Olefins Lienne et al.
PAHS Fielden and Packham
Davies et al.
Phenalenones Marshman
Saturatedaromatics IP PM-AZ
Saturatedaromatic types Apffel and McNair
Cookson et al.

364 Chapter 12
TABLE 12.1 (continued)
Davies et al.
Hazlett el a1
Diesel exhaust particulates

Naphtho[8,1,2-abc]coronene Jinno et al.
Nitrated PAHs Paputa-Peck et al.
MacCrehan e f al.
Fuel oil
Benz[a]pyrene Tomkins and Griest
Lubricating oils
Additives (over 50) Musha et al.
Musha et al.
Aromatichon-aromatic fractions ASTM D 2549
Renz[a]pyrene Saito
Furfural DeSanzo et al.
Naphthalene/phenanthrene Mazzeo et al.
PAH fractions Palmentier et al.
Ostman and Colmsjo
Polychlorinated biphenyls DeSimone et al.
Saturates/aromatics IP 368
Saturatedaromaticsipolars ASTM D2007
Pei et al.
Pei and Hsu
Sulphonates Bear
ASTM D3712
Sulphurized alkylphenols Chen and Nero
V1 improver Blanco-Gomis et al.
Zinc dialkyldithiophosphates Fodor and Newman
Heavy oils
Olefinic fractions Yamamoto and Akutsu
PAH fractions Coulombe and Sawatzky
Saturates/aromatics/PAH/resins/asphaltenes,
etc. Lancas et al.
Saturates/naphthenes/alkylaromatics/thiophenes Hsu et al.
Hsu et al.
Bitumen
PAH fractions Coulombe and Sawatzky
Refinery effluent
PAHs Symons and Crick
able and less than 87% for 4-6 ring PAHs. Saito [25] determined benz[a]pyrene
in lubricating oils and greases with fluorescence detection after an alumina
clean-up; precision was reported as 6 7 % RSD with a detection limit of
3.95 nglg.
HPLC has also been used purely as a fractionation technique for PAHs. Cou-
lombe and Sawatzky [26] applied this method to bitumens and heavy oils and
determined PAHs in the various LC fractions by GC. Palmentier et al. [27] em-
ployed a semi-preparative scale fractionation followed by GC-MS. Ostman and
Colmsjo [28] prepared PAH fractions from crude oil and used crankcase oil by
elution from short silica columns followed by an automated backflushed Bonda-
pak-NH, HPLC system. Individual PAHs in the final fractions were quantified
by GC. Detection limits were in the order of 1 ppm from a 10-15 mg sample
using GC-FID or 0.1 ppm by scaling up the initial silica clean-up.
Mazzeo et-al. [29] detected PAHs as quinones by oxidizing them with CeN.
Reductive mode electrochemical detection was employed to achieve detection
limits in the order of ppb. Chromatography was performed on an ODS column
using propan-2-01 /phosphate buffer as eluent. These authors applied the above
system to the analysis of naphthalene and phenanthrene in a motor oil.
A proposed Institute of Petroleum standard method, IP PM-BN, also exists for
the determination of PAHs in petroleum, coal and shale oil products. Detection
limits of 0.1% d m of total, and 0.1 mg kg-1 of individual PAHs are quoted. The
method uses open, gravity feed silica columns to produce a PAH extract which is
further separated by HPLC on a 5,um particle Spherisorb amino column or
equivalent. The isolated 4-6 ring fraction is then run on a Sephadex LH20 parti-
tion column in order to separate alkylated PAHs from the parent PAHs. These
parent PAHs are individually determined by GC. Precision has yet to be estab-
lished.
12.4.1.2 Other indigenous compounds
Nitrated PAHs in diesel engine exhaust particulates have been examined by

Paputa-Peck et al. [30] and MacCrehan et al. [31]. Paputa-Peck employed a
normal phase HPLC fractionation of methylene chloride extracts. Determination
of individual nitrated PAHs was by GC with a nitrogen-phosphorus detector or
by GC-MS. MacCrehan separated methylene chloride extracts by RP-HPLC and
compared voltammetry, amperometry and fluorescence for direct detection of
individual compounds. Diesel particulates have also been examined by Jinno et
al. [32] for naphth0[8,1,2-abc]coronene using reversed phase separations and
multichannel W detection.
High molecular weight heterocyclic nitrogen and sulphur compounds have
been studied by Borra et al. [33] and Andreolini et al. [34]. These authors used

366 Chapter 12
highly efficient capillary LC columns and a combination of direct diode array

fluorescence detection or fraction collection and mass spectrometry to examine
solvent refined coal distillate, syncrude and shale oil samples.
Polycyclic aromatic nitrogen compounds (aza-arenes) in Arabian Light crude
oil have been examined and identified by Grimmer et al. [35]. These authors
used a TLC/HPLC isolation scheme with separation and identification of indi-
vidual compounds by GC. Aza-arenes, anilines and alkyl aromatic amines in
gasoline, kerosene and diesel fuel have also been studied by Nondek and
Chvalovsky [36] using two different charge transfer columns, 3-(2,4-
dinitrobenzene su1phonamido)propyl silica and 3-(2,4-dinitroanilino)propyl sil-
ica. A comparison of five different charge transfer columns for the separation of
aromatic compound classes from a crude oil distillate sample and other fossil
fuel samples has been made by Thompson and Reynolds [37].
Phenalenones such as 7H-benz[d,e]anthranen-7-one,benzanthrone and methyl
phenalones have been quantified in middle distillates by Marshman [38] using
silica reversed phase separation and UV detection at 400 nm. Detection limits
quoted are typically in the region of 0.2 mg I-*. Dibenzothiophene has been
quantified in crude oils by Rebbert et al. [39] and by Christensen and White
[40]. The former authors employed HPLC to fractionate samples for GC/FPD
quantification. In contrast, the latter authors used a novel LC-tandem MS system
to separate and unambiguously identify dibenzothiophene directly. Indigenous
phenols in crude oil have been examined by MacCrehan and Brown-Thomas
[41] with detection limits of less than 100 ng/g. These authors used alkaline sol-
vent extraction of the oil, solid phase purification of the extract and RP-HPLC
with electrochemical detection.
HPLC has even been applied to asphaltenes in order to assist the determina-
tion of an average molecule. Monin and Pelet [42] used size exclusion and a
range of bonded phase columns to fractionate such samples after selective disso-
lution in a number of solvents.
12.4.I . 3 Additives and contaminants
The anti-oxidant 2,4-dimethyl-6-tert-butyl phenol has been quantified by nor-
mal phase isocratic HPLC with UV detection and is the subject of an IP Standard
Method, IP343. The method allows a number of columdmobile phase combina-
tions and, in our experience, is robust and precise. Published repeatability and
reproducibility at the 200mg/l level are 2.61 and 6.56, respectively. Some
homologues and isomers of this compound may also be separated using varia-
tions in mobile phase composition. The same compound has also been quantified
with electrochemical detection by Hayes and Hillman [43].
Alkyl nitrate cetane improvers in diesel fuel have been determined by
Schabron and Fuller [44]. Normal phase LC on silica coupled with variable
wavelength IR detection was used to separate and quantify amyl, hexyl and octyl
nitrates. Recovery, accuracy and precision quoted were good and detection limits
of 0.05 and 0.01 vol.% are given for amylhexyl and octyl nitrates.
Up to 50 lubricating oil additives have been separated and retention times de-
termined by Musha et al. [45,46]. These authors used both normal and reversed
phase columns with UV detection at the maximum absorbance for each com-
pound.
Furfural has been determined in lubricating oils by Di Sanzo et al. [47]. A
5 p m ODs-silica column with an eluent of 70:30 watedmethanol and W detec-
tion at 280 nm gave a recovery >95%, good precision, and good agreement with
a bisulphite extractionKJV method. Samples for HPLC were pre-extracted with
methanol and cleaned-up with a C18 silica cartridge prior to determination.
Synthetic and indigenous sulphonates, including alkyl benzene sulphonates, have
been separated and quantified by Bear [48]. This author evaluated the evapora-
tive light scattering detector in the analysis of a wide range of surfactants and
concluded ...a uniform linear response for each class of surfactant, with detec-
tion limits in the low nmole range. In particular, the detector response was re-
ported to be independent of the alkyl chain length and the degree of aromatic-
ity with respect to alkyl benzene and alkylaryl petroleum sulphonates. Columns
and mobile phase varied according to the application and samples were analyzed
after dilution of the parent product. A diode array UV detector was also used in
series with the ELS detector. Standard deviations of all the analytes were less
than 1%.
Sulphurized alkylphenols have been separated from reaction side products and
base oil on a normal phase, y-cyclodextrin silyl column with gradient elution and
evaporative light scattering detection by Chen and Nero [49]. Individual fraction
from the separation were also characterized by mass spectrometry. Fingerprint
comparison between samples which passed and failed engine test specifications
are presented. The advantages of the ELSD over RI detection were stated by
these authors to be freedom from ambient temperature variation effects, minimal
baseline drift with multiple solvent gradients and a response which was mass
dependent rather than concentration dependent.
To illustrate the breadth of HPLC applications in the field of lubricating oil
additives, normal phase and reversed phase methods have even been applied to
the characterization of poly(styrene-alkylmethacry1ate)co-polymer viscosity in-
dex improvers of molecular weight up to 300 000 Da by Blanco-Gomis et al.
1501.
An Institute of Petroleum method exists for the determination of the coumarin
content of kerosene. This compound, 1,2-benzopyroneis often added to kerosene
as a marker for excise purposes. The method uses a silica column, a mobile
phase of 2% propan-2-01 in hexane or heptane and W detection at 274nm.

368 Chapter 12
Typical calibration range is 2-4 mg/l and at the 2 mg/l level, repeatability is
quoted as 0.06 and reproducibility 0.28.
12.4.1.4 Compound classes
The inherent normal phase separation mechanism (adsorption) has the ability
to separate complex mixtures of hydrocarbons according to degree of unsatura-
tion. As such, it has been widely exploited in the characterization of petroleum
products with respect to the saturate, monoaromatic, diaromatic, triaraomatic,
polar and (to a lesser extent) olefin content. Ongoing development of bonded
normal phases has largely been aimed at achieving cleaner cut-offs between
compound classes, most notably by the use of substituent groups which separate
by charge transfer mechanisms with the aromatic nuclei of the sample compo-
nents. Products are often quantified in terms of the mass or volume fraction of
each compound class present, and further separation of individual components
within any class is either not possible or unnecessary.
No fewer than five standard IP methods of this type exist covering aviation
fuel, auto diesel, drilling mud oils, gasoils and lubricating oil base stocks. Two
distinct HPLC technologies and quantitation methods are employed, both with
isocratic elution.
Silica columns and backflushing are used to separate saturates from total aro-
matics in basestocks (IP 368) and gasoils (IP PM-AZ) with gravimetric quantita-
tion and in aviation fuel (IF PM-AT) with RVUV detection and quantitation of
total aromatics and naphthenes. In all three cases, saturates elute through the
columns unretained and aromatics (with or without olefins) are backflushed off.
In auto diesel and drilling mud oils, two amino bonded phase columns are
used to separate mono, di and triaromatics with RI detection and external stan-
dard quantitation (IP 391 and IP PM-AY). The main concern in these last two
methods is that the external standards chosen are individual compounds, whereas
the actual sample components present in each class are many and varied. Detec-
tor response factors between sample and standard can therefore vary and will be
composition dependent.
A wide range of petroleum products and crude oil have been characterized by
HPLC and it is best to consider each one in order of product type.
Crude oil has been characterized by Welch and Hoffman [SO]. These authors
used an on-line microbore LC-GC-MS system with a 2,4-dinitrophenyl mercap-
topropyl silica LC column. The system employed a retention gap between the LC
and the GC columns and no attempt at quantitation was made. This article also
includes the analysis of JP-4 aviation fuel, isolating and identifying alkylben-
zenes, alkyltetralins, alkylbiphenyls, naphthalene and dimethyl naphthalenes.
Gasolines have been characterized by Apffel and McNair [52], Cookson et al.
[53], Mussi et al. [S4]and by Hayes and Anderson [SS] on an aminopropyl silica
HPLC and column liquid chromatography 3 69
column to separate alkene-free gasolines into saturates, monoaromatics, diaro-

matics, tri/polyaromatics and polar groups. Each group was quantified by an RI
detector into weight percentage abundance. Calibration samples were obtained
by fractionation of a fuel sample rather than by use of single pure compounds in
an effort to minimize compositional and RI response factor differences.
Both Apffel and McNair and Munari et al. used on-line HPLC-GCRID meth-
ods to analyse gasoline saturates, unsaturates, aromatics and polar compounds.
The latter authors employed a retention gap between the two chromatographic
systems and microbore HPLC columns. Hayes and Anderson used off-line
HPLC with a dielectric constant detector to achieve an accurate group type sepa-
ration and quantitation of gasoline with uniform response factors from the detec-
tor. The mobile phase was 2,2-dichloro-1, 1,1-trifluoroethane (Envron 123). The
individual fractions were then analyzed by GCMSD to identify components and
GC/FID to quantify them. The authors reported that spent Envron 123 can be re-
used several times without purification or easily redistilled on a continuous
basis.
Kerosenes have been characterized also by some of the authors previously
cited [51,53,55]. In addition, Haw et al. [56] used a propylamino silica column
with on-line NMR as the detector. Ln this case, the mobile phase was l,l,l-
trichlorotrifluoroethane with 2.5% deuterochloroform and 0.05% hexamethyld-
isiloxane as NMR reference. Each compound class (monocyclic and dicyclic
aromatics) could be given an average composition. The average composition of
the saturate fraction was, however, limited by problems in accounting for qua-
ternary carbon. Davies et al. [57] utilized the LC-retention gap-GCRID ap-
proach to a kerosene sample with microbore amino and silica glass-lined LC
columns in series with pentane eluent and backflushing. Unfortunately, the low
dead volume of detectors required for microbore LC precluded conventional RI
or dielectric constant detectors and thus direct quantitation of the saturate and
aromatic fractions prior to GC was not possible. The system was automatic and
clearly improved the analysis of the aromatic fractions.
Diesel and distillate fuels have been studied by all the methods described for
crudes, gasolines and kerosenes [52,53,57]. Silica and amino columns have been
used to separate diesel into saturates, olefins and aromatics with RI and/or U V
quantitation by Felix et al. [58]. Davies et al. [59] used the LC-GC technique
previously described, but with specific reference to polynuclear aromatics in die-
sel fuel. The chromatographic system described by these authors produced a
complete fractionation by compound class but in this study emphasis was placed
on the definition of a two-dimensional (LC versus GC) retention map for se-
lected PAHs. By comparison, the retention indices of aromatic compounds from
the diesel sample led the authors to conclude that naphthalene, phenanthrene and
their alkyl derivatives were the predominant aromatics present.
Refeences pp. 3 72-3 74

3 70 Chapter 12
The LC-NMR approach, previously applied to kerosene, has been extended to

diesels and distillates by Hazlett et al. [60], again culminating in the determina-
tion of average composition for each compound class. From LC-NMR data,
Caswell et al. and Swann found it possible to predict the physical properties of
diesel and jet fuels [61,62]. Multiple regression analysis was used for the corre-
lation of 13 LC-NMR parameters from each fuel with 17 physical properties
such as cetane number, distillation data, flash point, pour point, density, etc.
Thirteen of the 17 properties in reference [61] had correlation coefficients in
excess of 90% and seven were in excess of 95%.
Copper(li) and silver-modified silica columns have been prepared by passing
ammoniacal CuSO, through the column or by use of ammoniacal AgN03 during
packing by Lienne et al. [63]. With pentane or Fluorinert FC72 as mobile phase,
olefins could be separated from light and heavy distillates with RI and UV de-
tection.
Heavy hydrocarbons have been characterized by Hsu et al. [64,65] by on-line
LC-MS. It was reported that distinction could be made between naphthenoaro-
matics and alkylaromatics and also between aromatic hydrocarbons and thio-
phenes. The value of this kind of information for refinery processing is very
high.
12.5 PREPARATIVE HPLC AND COLUMN LIQUID

CHROMATOGRAPHY
12.5.1 Standard methods
The 1993 Annual Book of ASTM Standards [66] published by the American
Society for Testing and Materials lists six liquid column chromatography meth-
ods of relevance to the petroleum industry.
ASTM D13 19 is identical to the Institute of Petroleum, London method IPI 56
entitled Hydrocarbon Types in Liquid Petroleum Products by Fluorescent Indi-
cator Absorption. It is limited to samples boiling below 3 15C which are sepa-
rated by it into saturates, olefins and aromatics by elution through a silica col-
umn with 2-propanol under air or nitrogen pressure. Fluorescent dyes are added
to the top of the column which co-elute with the olefins and aromatics and serve
to mark the boundaries of each zone. The saturates front coincides with the wet-
ted front of the material passing down the column. The lengths of each zone are
measured at the end of the separation and these lengths are proportional to the
percentage of each class present in the sample. This test has been in use with
slight modifications for many years and is especially relevant to gasolines and
aviation kerosenes. Its main drawbacks are that it is time-consuming and opera-
tor intensive and that strict control of the silica gel quality is critical.
ASTM D2002 and D2003 are also silica gel fractionation methods used to
provide representative saturate fractions from low and high olefinic naphthas,
respectively. Again 2-propanol is used under pressure as the mobile phase.
ASTM D2001 uses a dual column system of Attapulgus clay and clay/silica
gel. Saturates, aromatics and polar fractions from oils with initial boiling points
greater than 260C are recovered from the columns. This analysis is often re-
ferred to as the Clay-Gel method. Once again it is a time-consuming and la-
bour intensive technique. This type of separation has been the subject of HPLC
development by Pei et al. and by Pei and Hsu [67,68].
ASTM D2549 furnishes aromatics and non-aromatics for further analysis by
mass spectrometry. In this method, up to 10 g of oil boiling between 232 and
538C are separated on a column of bauxite and silica gel. Pentane is used to
elute the non-aromatics and the aromatics are eluted with successive portions of
diethyl ether, chloroform and ethanol.
ASTM D3712 uses a silica gel column with chloroform and ethanol to sepa-
rate diluent oil from petroleum and synthetic sulphonates. The average molecular
weight of the sulphonate is then determined by ashing the recovered material.
Metal sulphonates are first hydrolysed to sulphonic acids and converted to so-
dium sulphonates prior to the separation. Incompletely hydrolysed samples do
not separate well and, as the analysis is again labour intensive, care must be
taken in the hydrolysis step to avoid a considerable waste of effort. This method
is identical with IP Method 369.
12.6 INDIVIDUAL PUBLICATIONS
Zinc dialkyl dithiophosphate additives (ZDDPs) in finished lubricating oils

have been determined on 183 cm X 0.78 cm glass columns packed with 37-
75pm silica by Fodor and Newman [69]. The alkyl/aryl ratio of the ZDDPs in
the recovered methylene chloride fraction was determined by IR spectroscopy
against calibration solutions of the pure compounds. The authors concluded that
losses incurred in the chromatography limited the method to that of an estima-
tion rather than a quantitative determination.
Yamamoto and Akutsu used a preparative scale silica gel column to separate
saturates and aromatics and a 60 A silica/lO-20% w/v silver nitrate column to
separate saturates from alkenes. The argentation column could produce a 2 g
fraction of alkenes from a column packed with 80 g of argentated silica. Typical
samples included heavy distillates from thermal cracking [70].
De Simone et al. [71] obtained polychlorinated biphenyl extracts from petro-
leum samples by means of a combination of gel permeation and silica microcol-
umns. The PCB concentration in the extract was determined by GC with electron
capture followed by mass spectrometry to confirm structures.

3 72 Chapter 12
A mixed heavy end sample was separated into fractions with a 50 x 1 1 cm

silica gel Si60 column by Lancas et al. [72]. Two grams of the sample were
mixed with silica in a precolumn and single solvents or binary/tertiary mixtures
used to fractionate it. Six solvents in six mixtures of increasing eluotropic
strength gave saturates, monoaromatics, diaromatics, triaromatics, polynuclear
aromatics, resins, asphaltenes and asphaltols. Typical recovery is quoted as bet-
ter than 90%in most cases, with an RSD of 1.2%.
12.7 FUTURE TRENDS
A number of more selective column mechanisms are beginning to find appli-

cation in petroleum analysis. Most specifically, the range of selectivities now
commercially available in normal phase charge transfer columns such as DNAP,
TNAO and TNAP columns are allowing a more precise definition of aromatic
type cut point. As the industry has a continuing need for more precise total aro-
matic and aromatic type quantification, it is expected that the use of such col-
umns will increase. Similarly, the separation of functionalized bad actors from
a range of hydrocarbon products may prove to be accomplished by anion and
cation exchange columns which are now also commercially available.
Undoubtedly, the single most useful advance in detector design for the petro-
leum industry has been that of the evaporative mass detector. This detector will
find increasing use in the field for two reasons. Firstly, the operation of the de-
tector necessarily results in volatile sample matrices being evaporated along with
the HPLC mobile phases used in petroleum applications. This may actually
prove to be an advantage in the analysis of some gasoline additives. Secondly, as
heavier products will not suffer the same fate as gasoline/naphthas, characteriza-
tion of such samples can take advantage of the detectors true linearity and com-
position independence. These characteristics are unique in such a robust and
relatively inexpensive device.
Finally, LC-GC is still waiting for an enterprising manufacturer to develop a
truly turnkey system. Numerous applications of this hyphenated technique al-
ready exist which should be transferable. The analysis of oxygenates in gasoline
at percentage and at trace levels may yet prove to be the application which
arouses a sufficient volume of interest to be commercially viable.
12.8 REFERENCES
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5 N.A. Parris, Instrumental Liquid Chromatography, Elsevier, Amsterdam (1984).
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Dekker, New York. (1979).
8 R.P. W. Scott, Liquid Chromatography Detectors, Elsevier, Amsterdam (1977).
9 A.T. James, J.R. Ravenhill and R.P.W. Scott, Chem. Inst., 746 (1974).
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Chromatogr. 99 (1974) 13.
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12 P. Arpino, B.G. Dawkins and F.W. McLafferty, J. Chromatogr. Sci., 12 (1974) 574.
13 W.M.A. Niessen and J. van der Greef, Liquid ChromatographyMass Spectrometry, Marcel
Dekker, New York (1992).
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15 S.D. Woodruff and E.S. Yeung, Anal. Chem., 54 (1982) 2124.
16 D.L. Ford and W. Kennard, J. Oil Colour Chem. Assoc., 49 (1966) 299.
17 D.L. Ford and W. Kennard, Aust. Pat. Appl. No. 33406.
18 J.M. Charlesworth, Anal. Chem., 50 (1978) 1414.
19 Standard Methods of Analysis and Testing of Petroleum and Related Products, 1993, Institute
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20 E. Katz and K. Ogan, 5th Int. Symp. on Chem. Anal. Biol. Fate Polynucl. Aromat. Hydrocar-
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21 J. Grimalt and J. Albaiges, Afinidad, 40(385) (1983) 223.
22 B.A. Tomkins and W.H. Griest, J. Chromatogr., 386 (1987) 103.
23 P.R. Fielden and A.J. Packham, J. Chromatogr., 479 (1989) 117.
24 R.K. Symons and I. Crick, Anal. Chim. Acta, 151 (1983) 237.
25 T. Saito, Bunseki Kagaku, 39 (1990) 21 1.
26 S. Coulombe and H. Sawatzky, Fuel, 65 (1986) 552.
27 J.P.F. Palmentier, A.J. Britten, G.M. Charbonneau and F.W. Karasek, J. Chromatogr., 469
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28 C.E. Ostman and A.L. Colmsjo, Fuel, 68 (1989) 1248.
29 J.R. Mazzeo, I.S. Krull and P.T. Kissinger, J. Chromatogr., 550 (1991) 585.
30 M.C. Paputa-Peck, R.S. Marano, D. Schuetzle, T.L. Riley, C.V. Hampton, T.J. Prater, L.M.
Skewes, T.E. Jensen, P.H. Ruehle, L.C. Bosch and W.P. Duncan, Anal. Chem., 55 (1983)
1946.
31 W.A. MacCrehan, W.E. May, S.D. Yang and B.A. Benner Jr., Anal. Chem., 60 (1988) 194.
32 K. Jinno, J.C. Fetzer and W.R. Biggs, Chromatographia, 21 (1986) 274.
33 C. Borra, D. Wiesler and M. Novotny, Anal. Chem., 59 (1987) 339.
34 F. Andreolini, C. Borra, D. Wiesler and M. Novotny, J. Chromatogr., 406 (1987) 375.
35 G. Grimmer, J. Jacob, K.W. Naujack and D. Schneider, Erdoel Kohle, Erdgas, Petrochem.,
38(2) (1985) 82 (Synopsis 8504).
36 L. Nondek and V. Chvalovsky, J. Chromatogr., 3 12 (1984) 303.
37 J.S. Thompson and J.W. Reynolds, Anal. Chem. 56 (1984) 2434.
38 S.J. Marshman, Fuel, 5 (1990) 364.
39 R.E. Rebbert, S.N. Cheder, F.R. Guenther and R.M. Parris, J. Chromatogr., 284 (1984) 211.
40 R.G. Christensen and E.V. White, J. Chromatogr., 323 (1985) 33.
41 W.A. MacCrehan and J.M. Brown-Thomas, Anal. Chem., 59 (1987) 477.
374 Chapter 12
42 J.C. Monin and R. Pelet, Adv. Org. Geochem. Proc. 10th Int. Meet., Wiley, London, (1983)
pp. 839-846.
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48 G.R. Bear, J. Chromatogr., 459 (1988) 91.
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50 I>. Blanco-Gomis, J.M. LeBourgeois and R. Rosset, Chromatographia, 3 l(1-2) (1991) 71.
51 K.J. Welch and N.C. Hoffman, J. High Resolut. Chromatogr., 15 (1992) 171.
52 J. Apffel and H. McNair, J. Chromatogr., 279 (1983) 139.
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E.R. Adlard (Ed.), Chromatographyin the Petroleum Industry
CHAPTER 13
Modern data handling methods
N. Dyson
Dyson Instruments Lid., Hatton Lyons Industrial Estate, Hatton, Houghton-le-Spring,
Tyne and Wear DH5 Om UK
13.1 INTRODUCTION
The majority of current chromatographs use a data processor, either a stand

alone integrator or a computer-based system, to record, process and display
chromatograms and results. Prices range from just over $1000 for a single chan-
nel instrument, up to many thousands of doIlars for multi-channel computer sys-
tems.
The electronic technology of signal processing has advanced with the micro-
processor. Report preparation and distribution using spreadsheets, databases and
electronic mail can be both stunning and delightful, but the theory of peak meas-
urement used within these data processors is still that of the 1960s. Very little of
the research on peak separation is incorporated into commercial data processors,
overlapping peaks are still separated by perpendiculars and tangents. This meth-
odology has severe limitations and really ought not to be applied to the meas-
urement of all peaks, but it is. Current data processors are no more accurate than
a chromatographer of the 1960s working with a pencil and ruler, but they are a
lot more precise and very convenient to use. They can deliver results as soon as
the analysis is completed and it is very easy for the unwary professional or nov-
ice analyst to accept these results at face value.
The purpose of this chapter is to indicate how measurement errors arise,
highlight the limitations of modern data handling methods in coping with these
errors, and point to strategies of method development that minimize the difficul-
ties of data processing. It will steer analysts towards the correct balance between
good and sufficient method development and realistic use of integrators and
computers. Chromatographic data processors are not a substitute for a column or

376 Chapter 13
good experimental technique. Even the most expensive data processor is quite
capable of delivering a highly precise but totally inaccurate set of results.
13.2 THE ROLE OF THE DATA PROCESSOR
13.2.1 The modern integrator has four distinct roles
(i) Measure the detector signal. To take the signal as it emerges from the
detector and prepare it, without distortion or other loss of information,
for processing. The detector signal is analog in nature and is sampled by
an analog to digital ( A D )converter on the integrator input. This string
of sample readings is stored inside the integrator and processed by first
improving the signal to noise ratio and then extracting information on
the size of a peak and its retention time.
(ii) Calculate and report solute information. Analyte quantity and analyte
composition in terms of mass or concentration is measured from peak
area following calibration and the determination of response factors
from standard solutions. Height is an alternative measure of quantity
when the signaynoise ratio is poor. Analytes are identified by the reten-
tion time window in which they elute.
(iii) Control the experiment. Time-based programs control the mechanical
functions of the analysis: start the auto sampler, switch over valves, open
and close relays, etc. These programs can be in the chromatograph but
are often in the integrator. Similar programs optimize parameter settings
during the analysis. At the end of the experiment, the system is auto-
matically restored to its original configuration for the next analysis, or it
is re-configured for a different analysis.
(iv) Monitor the analysis results for feedback control, Additional informa-
tion from the integrator is used to monitor experimental stability. The
quality of the experiment can be assessed from additional data output:
The coefficient of variation of results over a series of experiments is a meas-

ure of instrument control. Variations in retention time indicate a lack of control
of mobile phase flow rates, column temperatures or gas leaks. The total peak
area count is a measure of injection volume. Its constancy (or otherwise) is a
measure of injection proficiency. Column condition can be monitored by meas-
urement of the area and height of a large isolated peak, (see Fig. 13.1). The ratio
(A/H)2 is an approximate measure of peak variance [ 13 and therefore column ef-
ficiency. Peak asymmetry ratio, BIA [2], defined in Fig. 13.2, is a test of column
suitability. If too many peaks are asymmetric, it may be better to switch to a col-
umn of more appropriate polarity.
Modern data handling methods 311
Fig. 13.1. Whatever the peak shape, the ratio of its area to height is equal to its width at some
height.
These are quantities that are readily calculated from existing data and can be
used as control diagnostics. The measurements can be used in a control feedback
loop to delay the next analysis until conditions are stable enough, or sound an
alarm if they are too bad for too long. It is possible to build systems which allow
automatic carrier gas and column switching followed by purge cycles, these re-
Fig. 13.2. Asymmetry ratio = BIA at 10% peak height [2].

378 Chapter 13
route samples to an alternative chromatograph if a fault occurs and nobody cor-

rects the problem in a specified period.
13.2.2 Lab economics
The sum of all the analysis times gives the utilization of a chromatograph.
This is used to estimate spare capacity and decide whether an additional chroma-
tograph is required. Utilization divided by the instrument costs provides the
capital cost per analysis. The numbers of similar analyses performed by different
analysts may be compared to judge who are the most efficient workers.
13.2.3 Prepare and distribute data, information and reports
Even stand alone integrators are not always stand alone any longer. Micro-
processor developments and data communications software for chromatography
[3] allows information generated by integrators to be incorporated into sophisti-
cated reports which show trends and make comments as well as give the basic
figures. These reports can then be distributed via electronic mail to a variety of
destinations. In some QC labs, the chromatographers role is to develop the
method and guarantee the quality of the analysis; he need not even see the
analysis results or be asked to comment upon them [4].
13.3 LIMITATIONS OF THE DETECTOR SIGNAL
It must not be assumed that the detector signal accurately reflects the profile
of the injected solute, yet it is clearly meaningless to measure the detector signal
unless it does. The signal is not representative of the solute unless the following
conditions apply.
13.3.1 Injector fractionation is uniform
Any injection port splitting of solutes must split each component species in
the same proportion as all others, i.e. there must be no discrimination. This sim-
ple requirement is very hard to achieve and has haunted the design of WCOT
injection ports. The multiplicity of existing designs implies that no single device
has yet got it right for all types of solutes. Although improvements in injector
design have been made and are continuing, it still requires a competent analyst to
produce good quantitative results using WCOT columns,
Modern data handling methods 3 79
Splitting carrier gas streams into two columns or detectors usually creates
fractionation problems.
13.3.2 All solutes of interest have eluted
Columns sometimes retain part of the sample after the end of the analysis,
which is why many need reconditioning regularly. The peaks which purge during
reconditioning were possibly .weighed as part of an earlier analyte but, clearly,
they did not appear in the report of that analysis.
13.3.3 Column resolution is adequate
If solute peaks are not resolved sufficiently they cannot be measured accu-
rately. The transition from packed to capillary columns showed just how well
peaks can conceal other peaks. Increased column performance does resolve
peaks and presents them for measurement, but the analyst can reach a point
where there is too much resolving power, where a single component of interest
splits into its isomers and these have to be rounded up and added together again,
if they can be identified, to give the required result.
13.3.4 All solutes of interest are detected
Solutes that produce zero response from a detector cannot be measured: FIDs
cannot detect water or inorganic gases, for example. Selective detection turns
lack of response into a virtue although the analyst must be very careful that his
quantitation does not deteriorate because some solutes are measured at the bal-
ance but not at the detector.
13.3.5 Signallnoise ratio is high enough
A solute peak must be clearly distinguishable from the background noise.

ACS guidelines [5] state that a peak cannot be clearly recognized unless the S/N
ratio is at least 3: 1 . The mininium ratio for quantitation is 10:1, although at this
ratio, the results are not expected to be particularly good.
In practice, what chromatographers regard as a noisy baseline has a S/N ra-
tio of about 25 or 30:l and there are theoretical reasons for defining a good
baseline as having a S/N ratio of 1OO:l or better [6] because when the signal is
this clean, the data can be used. for peak modelling.
3 80 Chapter 13
Fig. 13.3. Detector non-linearity effectively folds over the peak top.
13.3.6 Detector is linear
Saturation of the detector electronics is a built-in safety device to prevent the

electronics fiom damaging themselves. When the amplifier saturates, which it
can do on any of its range settings, it stops transducing the cell signal into an
output representative of the passing solute.
Before saturation, the relationship between analyte and detector signal
goes non-linear and gives a distorted measure of the solute. In the non-linear re-
gion, the peak top can be thought of as folding over (see Fig. 13.3). In extreme
cases when the top folds over completely, it can appear that there are two peaks
~71.
Until all of these factors have been tested and their associated problems over-
come, all chromatograms must be considered suspect sources of information no
matter how repeatable the results are.
13.4 DETECTOR SIGNAL PROCESSING
13.4.1 Improving signal to noise ratio
Detector signals always contain noise (unwanted frequencies) from chemical

and electronic sources. Noise hinders signal measurement and is therefore re-
moved before measurement begins. Detector sensitivity is increased by improv-
ing the signal to noise ratio (S/N). There are two basic techniques which data
processors currently employ: electronic filtering and software smoothing.
Modern data handling metho& 381
13.4.2 Electronic filtering
Electronic filtering uses inductive (LR) or capacitive (RC) components to ir-

reversibly filter high frequency noise. Inductive components are used in mains
power filters, capacitive components remove instrument generated noise. Time
constants are matched to the noise they are intended to remove. If they are too
small, they will leave too much noise, if they are too large they will remove the
noise but they will also distort peak shape; retention time and peak height are not
then reliable measures but the peak area remains constant. This distortion can
manifest itself in capillary chromatography where an old GC is used with a high
resolution column. The time constant of the detector amplifier may have been
built to accommodate packed column peaks and be too large for the narrower
WCOT peaks which are distorted in consequence.
13.4.3 Software smoothing
Software smoothing uses mathematical operations to remove noise. These al-

gorithms are either based on the moving window functions described by Savitsky
and Golay [8,9] or they use software filters matched to the noise fiequency [lo].
Smoothing is performed on the: stored signal data and, unlike electronic filtering,
is reversible as the original data is not destroyed. Moving window smoothing
assumes a high enough data sampling rate (about 100 samples per peak [ 1l]),
and not too much peak asymmetry otherwise the peak shape is further distorted
[12]. Even when sampling is fast enough, Savitsky4olay smoothing has the
disadvantage, that unless the noise is random, it moves high frequency noise into
the low frequency domain where it looks like baseline rubble and can interfere
with peak location or be measured in error as peaks [ 131.
At high S / N ratios, all smoothing techniques work well but trace analyses are
important, and this is where smoothing is least effective and most needed.
Smoothing algorithms are built into the integrator by the manufacturer and vary
from one manufacturer to the next. They are unknown to the analyst and their
validity cannot be easily checked.
13.4.4 Measurement of peak areas
One of the results of the rapid development of microprocessor-based instru-

ments is that all analysis data are stored and all signal processing is now post-
run, with the benefit of hindsight calculation. Data processors store the detec-
tor signal in digitized form. A single chromatographic peak is measured by the

382 Chapter I 3
trapezoidal rule after the peak boundaries have been located from the first and
second derivatives [ 1,147.
Location of peak boundaries is very sensitive to the S/N ratio and if they are
not located accurately, the baseline drawn below the peak will be placed incor-
rectly with consequential errors in area measurement [ 151.
Overlapping peaks, roughly similar in size, are separated for measurement by
dropping perpendiculars from the valley points between them. As resolution de-
creases, the valley points become harder to locate, and less accurate to use. By
the time a peak becomes a shoulder on another peak, its measurement is grossly
inaccurate.
When one peak is very much smaller than another, it is separated from the
larger peak by skimming a tangent beneath it and its area is measured above the
tangent. This measured area is underestimated in two ways: first, by constructing
a straight baseline where the true baseline (the tail of the larger peak) is curved,
and second, by using construction algorithms that depend on the peaks position
on the tail [1,16].
13.4.5 Baseline placement
Integrators and computers draw straight lines beneath peaks to simulate base-
line though at least one commercial system has simulated the baseline beneath a
tangent skimmed peak with an exponential fitted curve. The true baseline is the
detector signal in the absence of the measured peak (or peaks) and in tempera-
ture programmed analyses or where the measured peaks are skimmed from an-
other, the true baseline can be highly non-linear.
Where a group of overlapping peaks sit on a curved baseline, integrators place
a baseline beneath the whole group by means of the elastic band technique [l].
If the chromatogram is imagined to be a solid projection up and out of the chart,
the baseline constructed beneath a group will be the same shape as a piece of
elastic, stretched and placed around the underside of the group. This baseline
will touch the beginning and end of the group and some of the lower valleys in a
series of straight lines.
13.4.6 Mathematical separation (deconvolution) of peaks and integrator

developments
The mathematical separation of overlapping peaks using peak models has

been attempted since before the first electronic integrators were manufactured,
but, so far, without enough success to induce manufacturers to incorporate any
of the methods into data processors. The mathematical methodology of separat-

ing peaks falls into two broad categories:
1. selective extraction
2. resolution enhancement
13.4.7 Selective extraction
13.4.7.I Peak modelling

In modelling, peak shape models (e.g. Gaussian) are assumed and fitted to ex-
posed parts of a peak curve in order to allow the whole peak to be defined. This
is subtracted from the chromatogram envelope and the process repeated with
other peaks until all are removed and only baseline remains. The composition of
the analyte is then calculated from the subtracted peak areas. Many peak models
have been proposed: gaussian, exponentially modified gaussian (EMG), lorent-
zian, log normal and others [ 17-19].
The exponentially modified gaussian is bearing most experimental h i t [20,
2 11. Some success in deconvoluting simulated overlap of computer generated
EMG peaks has been reported [22], but real chromatograms remain a problem.
13.4.7.2 Signal subtraction
A form of selective extraction which does not depend on peak shape is the
subtraction of two stored signals (chromatograms). This has been adopted into
commercial integrators to remove baseline drift and/or highlight differences
between complex chromatograms. It is a development of the dual FID GC with
two identical columns to offset baseline drift on the fly. It creates purchase
economies because a single column GC can be used where a dual column GC
would have been bought; it extends specification because matched columns
could never subtract chromatograms, only baselines.
Two chromatograms are stored: one is the chromatogram with all peaks and
baseline drift, the other is the same chromatogram without sample injection, i.e.
the baseline and drift. Provided that the experimental conditions are stable, the
baseline should be identical in both experiments, and when one data set is sub-
tracted from the other, what remains are the peaks on a flat baseline. Constant
(and unwanted) events such as injection pulses or baseline spikes created by
valve switching are also removed.
Differences between complex chromatograms can indicate complex contami-
nation (more than one contaminant peak), or material degradation [22a]. The
difference chromatogram contains the errors of both chromatograms, but then, so
did matched columns.

3 84 Chapter 13
13.4.8 Resolution enhancement
In resolution enhancement, peaks variances are reduced until the peaks re-
solve sufficiently to be measured. Numerical techniques such as Fourier trans-
form and others have been used [ 10,23-261.
These approaches have generally disappointed because:
(i) There is not enough information in an FID or other single channel detec-
tor. With perfect deconvolution, a single peak can be resolved into its
components, and if this peak is pure, then deconvolution would count
the isomers, a number which can be checked independently. However,
an identically shaped peak of another species would have to yield an-
other set of isomers and a different number. GC MS and GC FTIR offer
more information, as does diode array detection but so far this is limited
to LC. The next challenge to the mathematical resolution of overlapping
peaks can be expected to come from multichannel detectors coupled to
computers powerful enough to untangle the data.
( i j ) Both the extraction and resolution approaches to peak separation assume
that peak shape is constant even if it does not conform to a specific
model. The quality of manufacture of chromatographs and columns is
not yet good enough to uphold this assumption. Repeated injections of
the same solute rarely produce identical chromatograms, but the situa-
tion is improving: Ghaoui has reported the successful improvement of S /
N ratio without significant peak broadening by ensemble averaging [27].
(iii) Much of the methodology for peak separation is adapted from spectros-
copy. When applied to chromatography and variously asymmetric peaks,
some methods have been shown to generate spurious peaks and to distort
peak shape further [ 12,131.
Over the years, resolution enhancement has been more successful than peak
modelling but the major improvement in resolution has been the introduction of
capillary columns, not better mathematics.
Until such time as all variables affecting peak shape and signahoise ratio are
brought under control, mathematical techniques will be successfully applied only
in limited circumstances, and integrator peak separation techniques will stay at
the perpendicular/tangent level.
13.5 MEASUREMENT ERRORS
13.5.1 Accuracy and precision
Integrators are very good at delivering repeatable results if the instrument

conditions are stable; unfortunately, these results might be totally wrong. This
highlights the difference between precision and accuracy.
Accuracy is how close results are to the right answer. This is what the ana-
lyst seeks, but if it is to be checked, the right answer must be known in advance
by some wholly independent means or it must have been predicted by theory.
Precision is a measure of how close experimental results are to each other and
they reflect experimental stability and control.
The difference between the right answer and the measured answer is called
bias, and bias is never revealed experimentally except when two sets of re-
sults relating to the same sample are in complete conflict and one has to be
wrong.
Imprecision is easily observed and measured as coefficient of variation (CV)
or relative standard deviation (RSD. Inaccuracy is observed only when there is a
good theoretical model to provide the correct value for comparison,
13.5.2 Errors created by noise
Noise determines the smallest quantity of solute that can be detected [ 5 ] . The
minimum detectable quantity is the smallest that can be clearly distinguished
from the background noise.
Noise creates errors around the base of measurable peaks by obscuring the
beginning and end of the peak making it difficult to judge where the baseline
should be drawn. It causes integrators to find valleys at the start and end of
peaks, delaying the real start of measurement and tripping end of peak measure-
ment too early. Area is lost. Noise valleys on top of peaks split the area meas-
urements so that several peaks are reported, with retention times within seconds
of each other.
Fig. 13.4. Noise causes premature termination of peak measurement and area loss from tail.

3 86 Chapter 13
13.5.3 Errors created by peak overlap
Overlapping peaks are separated by perpendiculars dropped from the valleys

between them to the baseline constructed below them, or by tangent skimming
depending on the relative peak sizes. These methods are known to be inaccurate
except in limited circumstances [28,29]. In the absence of better methods, inte-
grators and data processors continue to use them.
Perpendicular separation of two peaks is only accurate when both peaks are
the same size and shape and have equal response factors. When their response
factors are dissimilar, the valley height should be no more than about 5% of the
smaller peak height.
If one peak is larger than the other, perpendicular separation at the valley will
overestimate the smaller peak and underestimate the larger one. In other words,
the true cleavage point is not the valley bottom but some unidentifiable point up
the side of the smaller peak.
Tangent skimming is used to separate peaks when the relative peak heights
are about 10:1 [ 181. There is no theoretical justification for this transition point
[28] and some manufacturers offer a choice of size ratios. It is not only arbitrary,
it creates a discontinuity where the integrator switches from perpendicular to
tangent. As a result there can be large changes in measured area for small
changes in peak size.
13.5.4 Errors created by asymmetry
In itself, asymmetry is not a problem for integrators [ 11. Integrators measure

peak areas by the trapezoidal rule which makes no assumptions about shape. In
theory a highly asymmetric peak on a flat, noise-free baseline is measured with
the same accuracy as a symmetrical peak.
The problems caused by asymmetry are that it compounds and increases the
errors of noise and overlap, and that it is not a constant phenomenon but can
vary with peak size and ftom peak to peak within the same chromatogram.
Varying asymmetry continually frustrates peak modelling.
Tailing on a noisy baseline causes the integrator to terminate peak meas-
urement early as the tail disappears and becomes indistinguishable from the
noise [30].
Asymmetry occurs mostly at the base of peaks. It spreads peak bases and in-
creases the overlap between neighbouring peaks. Perpendicular separation of
asymmetrical peaks does not account for the unequal contribution of each peak
to the other.
Variable asymmetry invalidates comparisons of peak height within the same

chromatogram unless careful calibration has been made. If all peaks were the
same shape and varied only in size, overlap correction factors could be success-
fully applied to areas and height would be a simpler measure of quantity.
13.5.5 Transmission of systematic errors through large peak groups
Overlap and asymmetry can conspire to create systematic errors which

transmit themselves through the group, creating large measurement errors in un-
expected places [3 11.
The errors created by the use of perpendicular separation of two overlapping
peaks are observed in the individual peak measurements but not in the total
measured area of the pair. This is because the area lost from one peak is trans-
ferred to the peak on the other side of the perpendicular. The loss from one is
equal to and cancels the gain in the other, or the sum of the two errors is zero. In
general, the sum of the errors, ei, in a group of n peaks caused by perpendicular
separation is equal to zero:
Ce,= O (13.1)
n
alternatively, the error in any one peak measurement, ep,is equal, and opposite in
sign, to the sum of the errors in all the other peaks in the group:
(13.2)
n-1
In large groups of peaks such as are common in the WCOT analysis of petro-
leum samples, the errors, even if individually small, can accumulate into a con-
siderable error. The implication of Eq. (13.2) is that there is little credibility in
the measured area of any single peak if it is part of a large group. It is therefore a
priority of method development to break up large groups of peaks into smaller
groups if they contain peaks of specific interest.
Asymmetry predominantly occurs as peak tailing rather than fronting. This
produces a systematic trend in the measurement errors; area is predominantly
transferred to the next peak on the tail. The last peak in the group has no peak on
its tail to receive the transferred error and must therefore accumulate all the sys-
tematic errors of tailing/overlap if Eq. (13.1) is to hold true. As a result, where
there are large groups of overlapping and asymmetric peaks, the error in the last
peak area measurement is very large.

388 Chapter 13
Meanwhile, the total peak area of the group will be measured accurately, and
the component peak areas will be measured with misleading precision.
** CALCULATION REPORT **
CH PKNO TIME AREA HE I GHT MK IDNO CONC KAME
1 1 0.496 5305231 835362 R 1
2 0.996 5294473 697597 V 2 0.998
3 1.496 5278366 599840 V 3 0.9949
4 1.996 5261428 527155 V 4 0.9917
5 2.495 5233851 472062 V 5 0.9865
6 2.995 5228614 42900 1 V 6 0.9856
7 3.494 52 16771 394619 V 7 0.9833
8 3.993 5206816 366638 V 8 0.9814
9 4.493 5 172876 343769 V 9 0.9751
10 4.991 5 190388 324724 v 10 0.9784
11 5.49 5 18150 1 308343 v 11 0.9767
12 5.989 5177132 294498 v 12 0.9759
13 6.488 5172920 28246 1 V 13 0.9751
14 6.986 5 169050 271991 V 14 0.9743
15 7.485 5 165786 262994 V 15 0.9737
16 7.984 6755953 255005 V 16 1.2735
_ __ _______ -___ ___ _
. -__
. -------
TOTAL 8501 1128 6666057 15.024
Fig, 13.5. These asymmetric peaks are the same size but note the measurement error of the last
peak.
30 1
25
z
20
Sum of Area Measurement Errors = 0
15
w
m
:
Y
10
0
a
5
-5 J
1 2 3 4 5 6 7 8 9 1 0 1 1 1 2 1 3 1 4 1 5 1 6
Peak Number in Group
Fig. 13.6. Error propagation across an asymmetric group (see Fig. 13.5).
7 a
Fig. 13.7. Negative baseline dips are a major source of measurement error.
Fig. 13.8. Baseline placement can be grossly wrong.

390 Chapter 13
13.5.6 Baseline construction errors
Baseline construction errors fall into two categories:
(i) where the integrator constructs a linear baseline even though the true
baseline, i.e. the detector signal in the absence of peaks, is non-linear;
(ii) where detector signal disturbances, especially those which give negative
dips to the baseline, cause the integrator to draw a baseline in the wrong
place.
Baseline disturbances, especially those involving deep negative dips, can give
rise to enormous errors, especially when the integrator drops perpendiculars to
separate peaks instead of using a tangent skim.
13.5.7 Errors of tangent/perpendicular peak splitting
The criteria for perpendicular and tangent peak separation depend on peak
size, shape and resolution. Different manufacturer's integrators and computers
use different algorithms and measure the same overlapping peaks in different
ways [32,33].
The measured area of a tangent peak depends on its position on the tail of the
larger peak [16]. Studies with synthetic chromatograms have shown that there
can be more than 25% difference in the measured area of a tangent peak half
way up the side of a peak compared to the same peak located at the end of the
peak's tail [3 11.
13.5.8 Ease of use
The very precise results reported by integrators can mislead the analyst into
believing they are accurate; however, integrators and computers are so conven-
ient to use and have such a high output of work that they cannot be discarded;
there is nothing to replace them.
13.6 CALIBRATION
13.6.1 Use of area or height for quantitation
Solute quantity is proportional to peak area [ 11. Height is a single, substitute

measurement for area that wastes most of the peak information and assumes
peak shape does not change from one analysis to the next. The choice between
area and height weighs experiment noise levels against peak shape.
Modern data handling methoh 391
Integrators are primarily area measuring devices [34]. If instrument time con-
stants are too large or the data sampling frequency is not high enough for the
peaks being measured, peak shape is distorted, and peak height changes but not
area. Peak asymmetry may increase as the column ages in use, in these cases
area is the more reliable measure.
Detector non-linearity affects the top of peaks where they are narrowest and
therefore height more than area. Moreover, the linear dynamic range for height
measurements is less than for area measurements [35,36], therefore area is pre-
ferred to height if detector linearity is in question.
If the signalhoke ratio is poor (<25: 1 approx.), noise will obscure the begin-
ning and end of a peak, making baseline placement erratic. Uncertainty at the
base of the peak, the broadest part, is where it matters most and creates the
greatest area error. A gain or loss of 2% at the base of a peak will result in a gain
or loss of 5% to its area [15]. Height is therefore more accurate and becomes the
preferred measure with noisy chromatograms.
13.6.2 Calculations and response factors
The three standard calculations of chromatography are:

- area % or normalization
- internal standard
- external standard
These days further processing of results is possible by exporting them into
computer spreadsheet files and special maths packages where they can be
worked endlessly into new reports or used to monitor trends. These are too nu-
merous to include here but they do conserve the analyst's time. It is important
to remember that the quality of these sophisticated methods is entirely depen-
dent on, and no better than, the original accuracy and precision of the measured
areas.
13.6.3 Limitations of the standard calculations
13.6.3.I Area %
The concentration of a solute, ci, is given by
A;Ri
c . =- x 100% (13.3)
' EAjRj
where Ai is the peak area of solute i, Ri is the absolute response factor of solute
i, defined as the quantity of solute to produce a peak of unit area.
392 Chapter 13
Concentration calculated from Eq. (1 3.3) is only accurate when all solutes are
eluted and detected and the detector is linear over the range of concentrations
measured. It also assumes the areas are measured accurately.
13.6.3.2Internal standard
The concentration of a solute, ci, is given by
(13.4)
where As is the peak area of an added internal standard, Ris is the relative re-
sponse factor of the solute to the standard (ratio of the two absolute response
factors), Wsis the weight of added standard and W,, is the sample weight.
The internal standard method compensates for variations in sample volume as
the added standard peak area varies correspondingly. It also indicates when sol-
utes are retained on column or have not been detected because the concentrations
do not then add up to 100%. Its shortcomings are that it is not always easy to
find a suitable standard or a convenient stretch of vacant baseline to place it, and
one solute has to be standard for a variety of other solutes of differing responses,
concentrations and perhaps peak shape.
I3.6.3.3 External standard
The external standard calculation compares peak areas under standard and
non-standard conditions:
c,
4
=-ws (13.5)
A S
A standard solution is prepared containing all the compounds of interest in the

unknown samples and analysed. Unknown samples are then analysed and the
solute concentrations obtained by comparison to the standard using Eq. (13.5).
Since solutes are compared against themselves, there is no need to use response
factors; they would be identical in Eq. (13.5) and would cancel.
This method relies on the injection volumes of standard and unknowns being
identical, which rules out manual injection techniques for the GC analysis of
liquids as they are too erratic. External standard method is best used with
valve/sample loop injection of gases (or in HPLC) where the sample sizes are
relatively large.
13.6.4 Calibration curves
Standard solutions are analysed over a range of concentrations to determine

response factors and to determine or compensate for detector non-linearity. Cali-
Modern data handling method 3 93
bration curves of area versus quantity are then calculated. If enough standard
solutions are analysed, the data are fitted to polynomials, which may turn out to
be linear.
Modern data processors either have all the standard calculation facilities built
into them, or they can be programmed to perform non-standard calcula-
tions. Some export the data into a computer which has the necessary software.
After entering details of the prepared standards and carrying out the analyses,
calculation of response factors and subsequent applications to unknowns is
automatic.
13.6.5 Empirical correction of analysis errors
There are many causes of peak measurement inaccuracy, yet calibration often
produces accurate assays even for analyses where noise, overlap and asymmetry
are clearly present and creating the inaccuracies about which this chapter warns.
It is reasonable to question whether these inaccuracies are genuine and if so how
can any analysis give reasonable results. The answer lies in the empirical nature
of the response factor. When a response factor is determined from bad chro-
matography, it combines the lme response factor with another empirical factor
which converts the measured result into the correct answer. This empirical factor
is a function only of that instrument operating under the prescribed analytical
conditions; if the sample, method or chromatograph are changed, the system
must be recalibrated. However, if conditions do not change, then the empirical
factor remains constant and the analysis results are accurate. The measured re-
sponse factor compensates for bad chromatography. This fortunate compensation
has its limits. Problems arise when analyses slowly deteriorate, columns become
contaminated, analytes change peak size and shape, resolution changes; then the
compensation may no longer work. Unfortunately there is no noticeable bound-
ary where the compensation ceases to work, where calibrated analyses drift into
error. The analyst can only avoid problems by calibrating regularly and by
keeping the analytical conditions as stable and reproducible as possible between
calibrations.
13.7 VALIDATION AND STANDARD CHROMATOGRAMS
13.7.1 Meaning of validation
Validation in chromatography means the systematic proof of the reliability of

the analytical method in terms of precision and accuracy [37]. The opportunities
for errors in peak measurement described earlier are just too many and varied to

394 Chapter I 3
accept results naively. All analyses should be validated by estimating the uncer-
tainty of each result. This specifically means calculating the accuracy error (i.e.
bias) in each result, and the coefficient of variation or relative standard deviation
of the experiment series, but calculating CV or RSD is not always done in prac-
tice and quantifying bias may be impossible by its very nature.
The emergence and adoption of quality schemes in chemical manufacturing
has created the demand to routinely calculate CVs or RSDs but the pursuit of
accuracy is an unending task and success is never guaranteed.
13.7.2 System suitability
System suitability is the verification that a chromatographic system remains

within the operating limits established during validation throughout the series of
experiments [37]. This means that once calibration has been made, the analyst is
confident that operating conditions do not drift between calibrations.
13.7.3 Validation and standard chromatograms
In an ideal world, the integrator should be independently validated and cali-

brated by means of an independent standard chromatogram of accurately known
dimensions. It can then be used to validate and calibrate the chromatograph and
measure unknown samples. In practice, the chromatograph is used to validate an
integrator or computer by injecting standard solutions and measuring them. The
calibrated integrator is then used to measure unknown samples on the chroma-
tograph. This interdependence means that systematic errors cannot be uncovered
and the measurement of unknowns can contain large inaccuracies.
With existing chromatographs, detector output during the analysis of a stan-
dard sample is not sufficiently repeatable or reproducible to be a good candidate
for standardization; in any case there are very few appropriate standards; a
typical lab sample remains the simplest and best.
Electronic signal generators and computer programs exist to generate chroma-
tograms of known dimensions [29], and although there are no agreed interna-
tional standards for them, their output is independent and fixed, and they offer
the preferred method for independent validation. They are also a very good
training aid for integrators.
Signal generators are independent of the chromatograph but have the disad-
vantage that they produce a single output chromatogram which bears no re-
semblance to any real samples that the lab measures. Computer programs are
better able to create a variety of independent standard chromatograms of known
Modern data handling metho& 395
dimensions, modelling real chromatograms. These can be measured and the re-
sults compared to the known answer. Accuracy as well as precision can be
checked on a synthetic chromatogram, relevant to the lab.
Historically, there have been several techniques for generating reproducible
signals including rotating drums with flashing peaks [38], tape recorders [39],
simple signal generators [ 121 and, more recently, storing real chromatograms and
creating computer simulated chromatograms [ 13,311.
Electromechanical devices always suffer from wear: magnetic tapes stretch
and have a limited frequency spectrum, motor speeds wander and component
specifications vary with tolerance and ambient conditions.
Synthetic computer-generated chromatograms are an improvement because
they are fixed data files which cannot degrade (although files can be deleted).
Real chromatograms stored inside the data system which measured them have
relevance and variety. They are fixed and can be referenced at will. Such chro-
matograms are not so reliable ,when transferred to another integrator to compare
processing as the transferred signal will differ from the original detector signal
by having passed through the first integrator. When the signal is transferred to
the data processor of another manufacturer, the processing algorithms are almost
certainly different [32,33].
Synthetic chromatograms based on realistic peak models such as the expo-
nentially modified Gaussian model, generated inside a computer and output
through a DIA converter open up the possibility of defining a standard chroma-
togram. It may prove impossible to agree upon a versatile international standard,
but analytical groups can agree among themselves on their own standards
which will be reproducible in different labs. The DIA converter is always subject
to component variations but these are offset by normalizing and ratioing peak
areas.
Comparisons of commercial integrators by the way they process a synthetic
chromatogram have shown that unless they are given relatively simple signals to
measure, they will not only process them incorrectly (as described by this chap-
ter) but also differently to each other [32,33]. Manufacturers of compounds
which attract tariffs and duties (perfumes, wines and spirits for example), may be
tempted to buy the integrator that gives the lowest measure and attracts the low-
est tax invoice irrespective of whether it is the most accurate. Legal cases which
hinge on chromatographicanalyses are another concern.
13.8 STRATEGIES FOR PE,QKMEASUREMENT
The strategy is to get rid of the problems systematically before the peaks are
measured.

396 Chapter 13
13.8.1 Noise
Noise is always present if the detector sensitivity is high enough, but sig-
nalhoise ratio can be improved by increasing the sample size and reducing the
detector sensitivity. If the column is regularly conditioned and kept in good
condition, there will be less unwanted bleed. Good quality carrier gas, hydrogen
and air will introduce fewer impurities into an FID flame and give lower back-
ground noise. The chromatograph will generate less noise if it is regularly serv-
iced.
13.8.2 Baseline drift
Baseline drift comes from poor experimental control. It is important therefore

to systematically root out the causes (which are just too numerous to list here).
Avoid temperature programming if it creates drift, minimize temperature ramps
if they cannot be avoided.
13.8.3 Peak overlap
Peak overlap is a fact of life for most chromatographers. It can never be got
rid of entirely. Nevertheless, it is important to maximize resolution of peaks of
interest and keep overlapping groups of peaks as small as possible on a baseline
that is as flat as possible. Alternatively use a selective detector to pick out only
the required peaks.
Sometimes a shorter analysis is preferred to optimizing the flow rate for best
column performance. When speed and throughput are important, the analyst
implicitly accepts less accurate results.
13.8.4 Asymmetry
Poor injection port design has been the major cause of off-column asymmetry
but this is rapidly improving; dead volume has largely been eliminated. Glass
inserts which restrict the expansive upstream flow of injected solutes remove
many of the problems. Off-column adsorption sites in the sample flow path, par-
ticularly septum shavings in the glass insert, must be removed.
On-column asymmetry is minimized by selecting a column of appropriate po-
larity although modern column manufacturing techniques give rise to good peak
shape in most cases.
An old chromatograph with a slow detector time constant will distort peak
shape. Please a salesman, throw it out and buy a new one.
13.8.5 Detectors
The linear dynamic range of detectors should be determined for all measured
analytes. It is important to note that detectors have different linear ranges for
different substances. It is possible to have a detector working linearly for one
component and non-linearly for another in the same sample.
When major peaks of interest go off scale, the analyst should occasionally in-
crease attenuation and check the tops of the peaks to confirm they have not been
clipped by the amplifier saturating.
13.8.6 Integrators
The analyst should know the limitations of integrators, read the manual and
learn how to program them. Give an integrator only those chromatograms which
it can measure accurately. A good chromatogram is one where solutes are well
resolved and reasonably sized for ease of measurement on a flat, noise-free
baseline.
13.8.7 Checking the analysis results
Results from an integrator or computer should never be accepted unquestion-

ingly.
13.8.7.1 Checking the chromatogram
A chromatogram is good enough to be measured if:
- it looks very similar to a standard or well known chromatogram
- there are the correct number of peaks, i.e. the right sample was injected
- the peaks are the right size, i.e. the injection volume was correct
- the noise level is acceptable; the column does not need conditioning and
earlier solutes have completely eluted
- the baseline is where it ought to be; the system had stabilized before in-
jection, and there are no dips afterwards
- baseline drift is acceptable or absent
- integration event marks are all present and correct

398 Chapter 13
The rule with event marks is: one event mark at every event (peak boundary
or valley) and no event missed. It suggests the measurement parameters were set
correctly and the peaks were integrated correctly.
13.8.7.2 Checking the analysis report
If the report is to be believed, all required peaks must be correctly identified.
If they are not, the integrator time windows are probably not set correctly, in
which case the wrong calibration factors will be applied when calculating peak
quantities.
Peaks must have the correct measurement diagnostic indicating that they were
measured as single peaks or separated from others by a tangent or perpendicular.
If they do not, it is, once again, an indication that the data processor is not pro-
grammed correctly.
13.8.7.3 Accepting the results
Finally, the actual peak areas and retention times must be repeatable within
previously accepted limits to show that experimental stability is good, and the
measured quantities must be within an accepted range to confirm that the prod-
uct is within specification.
13.9 REFERENCES
1 N. Dyson, in Chromatographic Integration Methods, Chromatography Monographs Series

(R.M. Smith, Series Editor), Royal Society of Chemistry (1990) ISBN 0-85186-587-9.
2 J.J. Kirkland, W.W. Yau, 1-l.J. Stoklosa and C.H. Dilks Jr., J. Chromatogr. Sci., 15 (1977)
303.
3 D.C. Nelson, V. Dauciunas, R. Lysakowski and M. Duff, Int. Lab., 32 (1992).
1 I. Scott, ICI Chemicals and Polymers, Personal Communication (1988).
5 D. MacDougall, Anal. Chem., 52 (1980) 2242.
6 T. Petticlerc and G. Guiochon, J. Chromatogr. Sci., 14 (1976) 53 1.
7 P. Deng and F. Andrews, J. Chromatogr., 349 (1985) 415.
8 A. Savitsky and M.J.E. Golay, Anal. Chem., 36 (1964) 1627.
9 J. Steiner, Y. Termonia and J. Deltour, Anal. Chem., 44 (1972) 1906.
10 D.L. Massart, B.G.M. Vandeginste, S.N. Deming, Y . Michotte and L. Kaufman, Chernornet-
rics: A Textbook. Elsevier, Amsterdam (1988) ISBN 0-444-42660-4.
11 S.N. Chesler and S.P. Cram, Anal. Chem., 43 (1 97 I ) 1922.
12 S.P. Cram. S.N. Chesler and A.C. Brown, J. Chromatogr. 126 (1976) 279.
13 M.R. Schure, J. Chromatogr., 550 (1990) 5 I .
15 A.B. Littlewood, Z. Anal. Chem., 236 (1968) 39.
I6 R.J. Hunt. J. High Resolut. Chromatogr. 8 (1985) 347.
17 S.T. Bake. in Quantitative Column Liquid Chromatography: A Survey of Chemometric
Methods, Journal of Chromatography Library, Vol. 29, Elsevier, Amsterdam (1984).
18 A.N. Papas, CRC Crit. Rev. Anal. Chem., 20 (1989) 359.
19 J. OlivC and J.O. Grimalt, 1. Chromatogr. Sci., 29 (1991) 70.
20 J.G. Dorsey and J.P. Foley, J. Chromatogr. Sci. 22 (1984) 40.

21 M.S. Jeansonne and J.P. Foley, J. Chromatogr. Sci., 29 (1991) 258.
22 C.P. Cai andN.S. Wu, Chromatographia, 31 (1991) 595.
22a E.R. Adlard and R.E. Davies, in Petroanalysis 81. Advances in Analytical Chemistry in the
Petroleum Industry (G.B. Crump, Ed.), Wiley, Chichester (1982).
23 R. Amino, in Advances in Chromatography, Vol. 15, Signal and Resolution Enhancement
Techniques in Chromatography, Marcel Dekker, New York (19??) Chap. 2.
24 R. Bracewell, The Fourier Transform and its Applications, McGraw-Hill, New York (1965).
25 Y. Hayashi, T. Shibazaki, R. Matsuda and M. Uchiyama, J. Chromatogr., 407 (1987) 59.
26 G.H. Rayborn, G.M. Wood, B.T. Upchurch and S.J. Howard, Am. Lab, 18 (1986) 56.
27 L. Ghaoui and L.D. Rothman, J. High Resolut. Chromatogr., 15 (1992) 36.
28 A. Westerberg, Anal. Chem., 41 (1969) 1770.
29 B.G.M. Vandeginste and L. DeGalan, Anal. Chem., 47 (1975) 2124.
30 B. Jover and J. Juhasz, J. Chromatogr., 154 (1978) 226.
31 N. Dyson, Int. Lab., 22(6) (1992) 38.
32 A.N. Papas and M.F. Delaney, Anal. Chem., 59 (1987) 54A.
33 A.N. Papas and T.P. Tougas, Anal. Chem., 62 (1990) 234.
34 G. Guiochon and C.L. Guillemin, in Quantitative Gas Chromatography for Laboratory Analy-
ses and On-Line Process Control, Journal of Chromatography Library, Vol. 42, Elsevier, Am-
sterdam (1988) ISBN 0-444-42857-7.
35 P.W. Carr, Anal. Chem., 52 (1980) 1746.
36 L.M. McDowell, W.E. Barber and P.W. Carr, Anal. Chem., 53 (1981) 1373.
37 T. Crispin and R. Wirtz, Chromatographia, 22 (1986) 434.
38 R.D. Johnson, J. Gas Chromatogr. 6 (1968) 43.
39 J.T. Shank and H.E. Persinger, J. Gas Chromatogr. 5 (1967) 631.
0 1995 Elsevier Science B.V.AU rights reserved 40 1
CHAPTER 14
Capillary electrophoresis in the petroleum

industry
Tim Jonesa and Gerard Bondouxb

a Waters Chromatography Division, Mill@ore (UK)Ltd, Winster House, Heronsww, Chester Business Park,
WrexhamRoad, Chester CH4 9QR, UK and bWaters Chromatography Division, Millipore S.A., 6 Rue Jean-
Pierre Timbaud, BP 307, 78054 St. Quentin-en- yvelines, France
14.1 INTRODUCTION
Capillary electrophoresis (CE) is a new, evolving analytical technique which

potentially offers considerable advances in the speed, efficiency and quality of a
number of application areas within the petroleum industry. CE has evolved fiom
classical electrophoretic techniques, the term capillary is used as the electro-
phoretic separation takes place in a fused silica capillary tube. Early electropho-
resis experiments in a glass tube were conducted by Lodge (1886) [l], who
monitored the migration of H+ in a tube containing a phenophthalein jelly.
Compton and Brownlee (1988) [2] have reviewed the last 100 years development
of CE. The use of small diameter glass tubes and silica capillaries has been re-
ported within the last 30 years by a number of authors [3-91. These efforts led to
the introduction of the first commercial instrument in 1989. While CE is there-
fore considered a new technique, there is a significant history of work to support
more recent efforts.
The number of publications since the introduction of commercial CE instru-
ments reflects the exponential growth of the technique in the last 5 years. The
first published applications concerned the separation of proteins and bio-
molecules in general (Fig. 14.1). The separation of other classes of compounds
such as pharmaceutical formulations, vitamins, inorganic ions, organic ions, car-
bohydrates and other natural molecules (as oppose to synthetic) have been ex-
plored. Even the separation of polymer particles by size has been demonstrated
[lo]; all the applications associated with classical electrophoresis can be

402 Chapter 14
5 10 15 20 25
Minutes
Fig. 14.1. Tryptic digest of cytochrome c (chicken). Analytical conditions: 60 cm X 75 p m capil-

lary, +15 kV, UV detection at 214 nm. Electrolyte A: phosphoric acid 30 mM, pH 2.0. Electrolyte
D: phosphate buffer 50 mM, pH 10.0. Electrolyte B creates a faster electro-osmotic flow and thus
shorter migration times.
achieved by CE. A number of liquid chromatography techniques can be trans-

ferred to CE. In common with sther new techniques, most of these applications
are confined to the research laboratory. There are, however, some control labora-
tories using CE for the routine analysis of ions and peptides.
A number of books are available on CE techniques [l 13 or on particular as-
pects of CE [ 12,131. The aim of this chapter is to give a short description of CE
such that its application to analyses within the petroleum industry can be real-
ized.
As in all electrophoretic methods, the separation of the analytes is achieved
using an electric field. The particular requirements for CE are illustrated in the
schematic of the instrument (Fig. 14.2). The system consists of a fused silica
capillary (in which the separation takes place), a detector, electrolyte and sample
vials, electrodes (usually platinum) and a high voltage power supply (capable of
supplying up to 30 kV). The sample is introduced into the capillary by a syphon
or other pressure system and the components separated by applying a potential
difference across the capillary. The components are monitored via the detector
which normally produces an anologue signal which can be recorded as in chro-
matographic techniques. The instrumentation required for CE is so simple that
most of the early work and indeed, a significant amount of current research, is
carried out successfully on home-built equipment.
The fused silica capillary used in CE has an internal diameter varying from 25
to ISOpm, and a length from a few centimeters to one meter. The use of small
diameter capillaries gives CE some significant advantages over other separation
Capillary electrophoresis in the petroleum industry 403
techniques. The most obvious is the efficiency of the separation. More than 106
plateslm have been reported. Compared with HPLC, there is little or no d i a s i o n
of the sample in CE; typical elution profiles are presented in Fig. 14.3.Low dis-
persion of the analytes results in very sharp peaks, high separation capacities and
good detection limits. Different kinds of detectors can be used to monitor the
migration profile, the detector is placed on-line and the separation is monitored
continuously. This can be contrasted with classical electrophoresis where the
separated species are monitored off-line once the separation is completed.
Data processing is the same as used in chromatography; calculation routines
have to be adapted for CE to obtain optimum results.
The similarities between CE and HPLC are obvious, for this reason CE is of-
ten considered as a combination of both techniques. This approach can be mis-
leading, especially when considering some of the peak-shape variations that CE
displays and it is more correct to consider CE as a separate analytical technique.
The terms given to liquid chromatographic techniques cover many different
methods, i.e. gel permeation chromatography (GPC), ion chromatography (IC),
reversed-phase chromatography (RPC)and fast-protein liquid chromatography
Fig. 14.2. Capillary electrophoresis configuration. 1, capillary; 2, electrolyte vials; 3, platinum

electrodes; 4, high voltage power supply; 6, sample carousel. Configuration A (normal configura-
tion): 5, detector (W or fluorimeter). Configuration B: 5, grounding device; 7, detector (ampero-
metric, or conductometric, or MS interface).
References pp. 4 2 5 4 2 6
404 Chapter 14
Fig. 14.3. Elution profiles in CE and HPLC. (A) Capillary electrophoresis; (B) HPLC. The electro-
osmotic flow generates a flat profile, compared to the profile of pressure driven flow. As a result
analyte zones in CE are very sharply defined compared with HPLC.
(FPLC). Different chemical approaches are used to obtain the separation in these
methods, but the instrumentation remains the same. Similarly, the term CE
refers to a group of different chemical approaches. The various separation meth-
ods, instrumentation and applications are described in the following pages.
14.2 SEPARATION TECHNIQUES
There are four main separation techniques used in CE. Other modes of sepa-
ration could be considered, but they generally belong to at least one of the four
main modes. These are:
- free-zone capillary electrophoresis (FZCE)

- micellar electrokinetic chromatography (MEKC)
- gel-filled capillary electrophoresis (GFCE)
- capillary isoelectrofocussing (cIEF)
Each separation technique is considered in turn and where appropriate is ref-

erenced to applications within the petroleum industry.
14.2.1 Free-zone capillary electrophoresis (FZCE)
The term FZCE is used to describe the separation that takes place in a capil-
Injector Detector
-
1 Neutral species (not charged) have electrophoretic
mobility
2 - Positively charged species will have a positive electro-

phoretic mobility. They will move in the capillary
with a speed higher than the speed of the electrolyte.
Usually kP<< po.
CI.0
3 - Negatively charged species will have a negative

electrophoretic mobility. They will move in the
capillary at lower speed than the electrolyte.
PO pep PaPP
L - =
+ L
Fig. 14.4. Migration of analytes in FZCE, po represents the electrophoretic mobility of the electro-
lyte.
lary filled with an electrolyte consisting of various aqueous salt solutions. The
composition of the electrolyte is the same in the capillary and in the two electro-
lyte vials (injection and detection sides). If different electrolytes are used at the
injection and detection ends of the capillary, the mode of separation is not FZCE
but isotachorphoresis ( not covered in this chapter). In FZCE, the electrolyte
contains no additives such as surfactants at micellar concentrations, or sieving
materials, but may contain material which can dynamically modifL the capillary
surface. It is important to appreciate that once a clear understanding of the
mechanisms of FZCE are gained, the other modes of CE separations will be
readily understood.
FZCE separations are governed by two parameters:
(i) mobility of the analyte,

(ii) electro-osmotic flow (EOF).
The analyte mobility represents the ability of a molecule to move in an elec-

tric field. The mobility of neutral molecules is zero, the mobility of charged
406 Chapter 14
molecules is proportional to their charge and inversely proportional to their

mass. The charge to mass ratio of an ion is therefore an important parameter in
CE separations. Both anions and cations can be separated in the same run.
Cations are attracted towards the cathode and under normal conditions, their
speed is additive to the electro-osmotic flow. Anions are attracted towards the
anode, but are normally carried to the cathode by the faster EOF. Neutral
molecules are carried towards the cathode by the EOF. The order of migration is
cations with the highest charge/mass ratio followed by cations with smaller ra-
tios. The neutral species migrate as one, their charge/mass ratio is zero. Finally
the anions with the lower charge/mass migrate earlier than those with greater
ratios.
The electro-osmotic flow within the capillary is a product of the electrical
field strength, the charge on the capillary surface, the ionic strength, pH and vis-
cosity of the electrolyte. In most cases, the electro-osmotic flow and the mobility
of the analyte ion are additive, although a number of well established applica-
tions of CE rely on differences between the electro-osmotic flow and analyte
mobility.
The mechanics of osmotic flow has been described in a number of publica-
tions [ 141 and is presented schematically in Fig. 14.5. Osmotic flow is created by
the presence of positive or negative charges on the interior walls of the capillary.
When using bare silica capillaries, the ionization of the silanol (SiOH) groups
occurs at pH 3 4 . The negative charges induce an accumulation of positive
charges (H,O+) near the walls. These hydroxonium ions may also be involved in
the solvation of analyte cations. When the voltage is applied across the capillary,
the positive charges are attracted by the cathode and they move (migrate) to-
wards it. The net effect is a movement of the electrolyte in the capillary in the
direction of the cathode. It is important to note that the EOF exists only if the
walls of the capillary are charged (e.g. if the electrolyte pH exceeds 3) and if the
internal diameter is small enough (less than 200 pm).
The strength of the EOF increases if the electrolyte pH is raised as a result of
the increased ionization (i.e. charge) of the silica surface of the capillary. Silica
Fig. 14.5. Electro-osmotic flow.

is fully ionized at pH 6.8, therefore increasing the pH of the electrolyte above

6.8 does not lead to an increase in EOF. It can, however, influence CE separa-
tions due to the effect on the ionic state of the analyte. The EOF can be affected
by modifying the state of the capillary walls. The addition of cationic surfactants
at millimolar concentrations can reverse the EOF. The positively charged surfac-
tant molecules form a layer at the surface of the capillary and effectively create a
positively-charged surface instead of a negative silica surface. Hydroxide ions in
the electrolyte will therefore associate with the coated surface. When the voltage
is applied, the hydroxide ions migrate towards the anode, i.e. the normal elec-
tro-osmotic flow is reversed. Similar results can be obtained by coating the capil-
lary with positively charged polymers. The stability of the coating, however, can
be adversely affected by the applied electric field, leading to poor reproducibil-
ity. Coating of the capillary with a neutral polymer can almost eliminate the
EOF, alternatively the injected samples may react with the walls and modify the
EOF. It is also possible to alter the ionized state of the capillary wall by coating
it with a weak acid. Positive or negatively-charged walls can be generated de-
pending on the pH of the electrolyte used. Further control of the EOF can be
gained by modifying the viscosity of the electrolyte; as the viscosity increases,
the EOF velocity decreases and this has been used to successfully separate sur-
factant species (Fig. 14.6).
Application work using FZCE must be defined by reference to the actual EOF
observed. The EOF can be determined using a neutral-charged marker molecule
(such as a UV-absorbing organic compound, e.g. formamide). Migration of a
neutral marker in a 75 p m X 60 cm capillary, at pH 7.0 and at an applied voltage
of 20 kV, will take approximately 10 min, giving a calculated value of the EOF
of 0.26pVmin. This value will vary, depending on the pH, nature and concen-
tration of the electrolyte and the applied voltage. The EOF can be compared to
the flow of solvent from an HPLC pump. It is necessary to maintain a constant
EOF to obtain stable migration times, just as it is important in HPLC for the
pump to give a stable flow for reproducible retention times. FZCE is a popular
mode of CE as it is relatively easy to predict the effect that modifications to the
analytical conditions will have on the subsequent separation. For this reason,
there are a large number of published papers and referenced applications.
Manipulation of the EOF and the factors that affect it are both the benefit and
the problem with FZCE. For the technique to be successful in a modern analyti-
cal laboratory, it is necessary to ensure that the EOF is correctly controlled. This
is especially true when analysing unknown samples; the operator must be wary
of the effect the sample analytes can have on the EOF. For this reason, there are
a number of capillary conditioning steps that can be used to minimize these ef-
fects. Rinsing the capillary with potassium hydroxide (normally at 0.5 M
strength) between samples is often used to remove sample components which
408 Chapter I4
__+____,
molecules molecules
B
Fig. 14.6. MECC with SDS addition. (A) Micelle structure. The interior of the micelle present hy-
drophobic properties. (B) The separation mechanism is a combination of FZCE and partition equi-
librium between the micelle and the electrolyte. The micelle has a lower mobility than the EOF
and therefore migrates towards the cathode.
may be bound to the silica surface. The reproducible analysis of quaternary-

amine-based detergents is greatly improved by rinsing the capillary between
samples with a rnethanolicKOH solution. In addition ,the electrolyte contains a
zwitterion at high concentration (1 M) to inhibit the adsorption of the amine to
the capillary wall El.51. An alternative to the use of zwitterions as a dynamic so-
lution is to use covalently coated capillaries. Unfortunately, these coatings are
not stable in use and consequently have short effective lifetimes.
14.2.2 Micellar electrokinetic chromatography (MEKC)
Neutral molecules cannot be separated by FZCE, they migrate at the same rate
as the EOF and will therefore be detected as a single unresolved peak. If differ-
ences in hydrophobicity exist between the various neutral molecules in the sam-
ple, it is possible to separate them using MEKC.
The addition of an anionic surfactant to the electrolyte at an appropriate con-
centration can result in the formation of micelles (Fig. 14.6a). The interior of the
micelle has a hydrophobic character while the exterior is hydrophilic. The neu-
tral analytes can be separated according to their affinity with the hydrophobic
part of the micelle. The technique has been extensively studied by Terabe and
co-workers [S, 16-20] who developed the technique working with phenolic com-
pounds as a model system, but they have looked at a broad spectrum of analytes
including chiral drug formulations [21]. MEKC uses an ionic micelle which will
normally have an electrophoretic mobility opposite to that of the EOF. The
mobility of the micelle is lower than that of the EOF such that adsorbed analytes
in the micelle will be detected, as they migrate to the detector at the cathodic end
of the capillary.
Separation is based on the analytes partitioning between the micellar phase
and the solution phase. Micelles form in solution when a surfactant is added to
water in a concentration above its critical micelle concentration. The most com-
monly used surfactant in MEKC is sodium dodecyl sulphate (SDS), which is an
anionic surfactant. A typical electrolyte contains SDS at 50 mM concentration
and a buffer such as sodium phosphate at pH 8.0.
The hydrophobic analytes in the sample are the most retained. This is similar
to what is observed in reversed-phase HPLC. In MEKC, the anticipated migra-
tion order is the same as the elution order in reversed-phase HPLC. One of the
limiting factors of HPLC is the minimum particle size of the column packing; the
smallest particles in common use today have a diameter of 3 p m . The dimen-
sions of the micelles, however, are much smaller and as a result, much better
efficiency can be obtained with MEKC.
As in HPLC, the addition of organic modifiers influences the separation by
altering the degree of adsorption of the analytes, depending on the amount of
modifier present. Increasing the amount of organic modifier in the electrolyte
will cause a decrease in the adsorption of hydrophobic analytes into the micelle
and a reduction in their migration times. It may also increase the viscosity of the
electrolyte, thereby reducing the EOF leading to an enhanced separation.
MEKC has been used for a wide variety of applications from the separation of
amino acid derivatives to the separation of preservatives (Fig. 14.7) or metal
complexes. The potential applications for MEKC include the majority of HPLC
reversed-phase applications, MEKC offers increased resolution at the possible
expense of sensitivity. It is important to note that while CE separations are more
sensitive in terms of the absolute sensitivity of analyte detected, HPLC is more
sensitive in terms of the absolute concentration of analyte. In a CE separation 1-
10 nl of sample are injected, this equates to picogram quantities of the analyte
410 Chapter I 4
4 5
1
5h -uv
t " " l ~ 5.00" l ~ " '

10.00l l l l '15.00
0.00
Mimrtea
Fig. 14.7. Separation of preservatives in the MEKC mode. Peak 1, phenoxy ethanol; 2, methyl-
paraben; 3, sorbic acid; 4, ethyl-paraben; 5, propyl-paraben; 6, isobutyl-paraben; 7, n-butyl-
paraben. Electrolyte: 25 mM phosphate, 25 mh4 borate; 5% acetonitrile (pH 8.0), 50 mM SDS.
Detection: UV 254 nm. Capillary 50 pm X 60 cm.
compared to the nanogram or microgram amounts injected via HPLC (where

routinely 5-50 pl of sample are injected).
Despite this obvious limitation, MEKC offers a number of advantages com-
pared with reversed-phase HPLC, for instance MEKC gives better separation
efficiency with a different selectivity (because the separation is based on both
free-zone and MEKC) from much smaller (nanoliter) injection volumes. There is
considerably less waste with MEKC due to the low consumption of solvents
compared with HPLC. MEKC should be considered, however, as a complemen-
tary technique to HPLC as gradient separations cannot at present be carried out
Capillary electrophoresis in the petroleum industry 41 1
by MEKC (although MEKC can replace the need for gradient HPLC [22]) and
the detection limits for HPLC (in terms of injected concentration) are better than
CE. HPLC is also usually a better choice if the analytes are only soluble in or-
ganic solvents (such as long-chain fatty acids). Although anionic surfactants
such as SDS are usually preferred, cationic surfactants such as CTAB can also
be used [23].
14.2.3 Gel filled capillary electrophoresis (GFCE)
This technique is a direct extrapolation of classical electrophoresis in that the

main separation mechanism is based on differences in analyte size as they mi-
grate through the pores of the gel-filled capillary. Cohen and Karger [24] used
polyacrylamide gels containing sodium dodecyl sulphate for very high efficiency
separations of proteins, polynucleotides and DNA-fragments. An example of the
excellent resolution available is illustrated in Fig. 14.8. An alternative approach
to binding a gel into the capillary is to incorporate cellulose or dextran deriva-
tives in the electrolyte. These viscous chemicals form a network in the capillary
and can act as molecular sieves. While the technique has been of considerable
value for the analysis of large biomolecules, the analysis of industrial polymeric
materials has not benefited from GFCE. Capillaries containing immobilized gels
Om 1
15 20 25 30 35
Minutes
Fig. 14.8. Separation of double stranded DNA using a sieving buffer. Electrolyte: 50 mM sodium
phosphate, pH 5.0 with 0.5% hydroxypropylmethylceliulose (4000 cP). Injection by electromigra-
tion (-5 kV for 5 s). Run: -10 kV. Capillary: 100pm X 1 m. Detection: UV at 254 nm. Sample:
1 kb DNA ladder (Gibco BRL) at 250 nglpl (Courtesy of Millipore).

412 Chapter 14
are expensive and fragile, they are easily poisoned by the non-selective adsorp-
tion of analyte molecules to the gel material. Sieving buffers are less fiagile and
less expensive but have not been sufficiently investigated to determine their po-
tential for the analysis of industrial polymeric materials.
14.2.4 Capillary isoelectric focusing (CIEF)
This technique is also derived from classical electrophoresis in which sub-

stances are separated on the basis of their isoelectric point or PI values. The
capillary is filled with an ampholyte solution which forms a pH gradient in the
capillary when the voltage is applied. The analyte molecules (almost exclusively
proteins) migrate to the point where they have a net neutral charge, at which
point they stop migrating. The focused zones of the sample can be eluted from
the capillary using pressure or the electro-osmotic flow of a different, non-
ampholytic buffer. This method provides very accurate determination of PI val-
ues and has a large sample-loading capacity; up to 1 p g of material can be loaded
onto the capillary, separated, and the purified fractions collected.
14.2.5 Instrumentation
As described briefly in the introduction to this chapter, CE systems have a

common architecture whatever their application. The different parts of the sys-
tem must be optimized to achieve reproducible analyses, ease of use, sensitive
detection and automatic operation. The main components of a CE system are
discussed in detail and the important parameters highlighted.
14.2.6 Capillary
Although CE is possible using glass or polymer capillaries, silica capillaries

are the most widely used as they give the best combination of properties for CE.
Alternatives to silica are usually explored in order to overcome difficulties with
the separation; in these circumstances it is usually more productive to investigate
an alternative technique. Alternatively, the chemistry of the electrolyte can be
adapted to dynamically coat the capillary surface. A considerable amount of ef-
fort is being put into improving the stability and nature of coating materials; a
number of significant developments are expected in the near future.
The dimensions of the capillary are important parameters. The internal diame-
ter is usually between 25 and 150pm. Smaller diameters can improve separa-
tions due to an increase in efficiency, but at the expense of detection limits

(injected volumes are smaller and the detector pathlength is shorter). The use of
larger capillary diameters can improve detection limits, but at the expense of the
separation. The recent development of Z-Flowcells for most commercial instru-
ments has improved detection limits , without the need to use large-bore capillar-
ies. The square of the diameter of the capillary is directly proportional to the cur-
rent generated in the capillary at a given potential. This limits the diameter of the
capillary since applying sufficient field strength to separate the analytes may
generate an excessive current if the capillary has a large internal diameter. This
can be manifest in two ways, the current generated exceeds the safety limits of
the system and it shuts down, or the heat generated by the current causes boiling
of the electrolyte and a break in the electrical continuity of the capillary. Before
either of these two points are reached, detector baseline instabilities will be ob-
served because of bubble formation.
The separation capacity of the capillary is proportional to its length. Keeping
other analytical parameters constant, increasing the length of the capillary de-
creases the electric field strength leading to longer run times.
14.2.7 High voltage power supply
The most important feature of the power supply is stability; as the pump of
the CE system, it is important to have a good-quality power supply. Most devices
will operate at up to +30 kV with a high degree of accuracy and precision. They
can run at constant voltage, constant current, or a mixture of both. Safety pre-
cautions are built into the CE system to prohibit inadvertent hazards. CE systems
cannot be considered spark-proof; this may limit the extension of their use in
hazardous areas such as oil platforms or refineries, unless they are operated in a
purged environment.
14.2.8 Temperature control
Applying a potential difference across the capillary and the resulting current
generates heat, this has to be dissipated to avoid boiling within the capillary. Dif-
ferent approaches are feasible, using either air flow across the capillary, or ex-
ternally cooled fluid circulation. With the latter approach, the capillary has to be
inserted into a cassette-type device to facilitate the coolant. Good temperature
control is necessary for reproducible migration times. A change of 1C causes a
2% change in ionic mobility (Fig. 14.9). Temperature control of the electrolyte
and sample compartments is also important. The selectivity of MEKC separa-

414 Chapter 14
3.0 1
I
2.9 J
carbonate
phosphote
fluoride
nitrate
nihite
sulfate
chloride
bromide
Fig. 14.9. Influence of temperature on the migration times of anions. Analytical conditions: electro-
lyte, 5 mM chromate, 0.5 mM CIA-Pakm OFM anion-BT. Courtesy of Millipore-Waters.
tions can be significantly altered by changing the temperature, while the stability
of temperature labile samples will be enhanced by low (4C) temperatures.
14.2.9 Injection
There are two modes of injection found on commercial instruments, hydrody-

namic injection and electrokinetic (or electromigrative) injection. A pressure
differential is used to load the sample into the capillary in the hydrodynamic
mode. This can be obtained by different means; gravity or air pressure at the in-
jection side, or vacuum at the detection side. If the capillary is held in a cassette,
pressure or vacuum has to be used for hydrodynamic injection. Air-cooled capil-
laries can be raised and lowered to facilitate gravimetric injection. Where direct
pressure or vacuum is used, it is important that there is accurate control of the
pressure and the sample vials are well sealed, so that reproducible injections can
be made. When using gravimetric techniques for injection, the capillary is placed
in the sample vial and elevated to a given height above the detection end of the
capillary. An aliquot of the sample will syphon into the capillary. The injected
volume is proportional to the pressure differential, sample viscosity, capillary
diameter and time. If any of these parameters cannot be controlled, the injections
will be irreproducible. Normally, the capillary diameter will stay the same, as
will the pressure differential if all seals are in good condition. One can manipu-
late the time of injection to facilitate loading; viscosity is more difficult to con-
trol, particularly if the sample matrix is subject to change. Temperature control
is important here because of its effect on the viscosity of solutions.
Injection volumes are very small, less than 10 nl is common, and should be
considered if sample homogeneity is not reliable. The resolution of the separa-
tion can be compromised if the injection volume is too large. It is difficult to de-
fine exactly the volume of sample that will disrupt a CE separation.
Electromigrative injection is used to selectively introduce sample components
into the capillary by generating a potential difference between the capillary and
the ions in the sample. The injection of negatively charged species is obtained by
making the end of the capillary positive with respect to the sample. The negative
ions migrate to the capillary where they can accumulate. Positively charged spe-
cies are injected by making the end of the capillary negative with respect to the
sample. Differing amounts of sample are injected by varying either the injection
voltage or the length of time the voltage is applied. Unlike hydrodynamic injec-
tion, electromigration is a selective injection technique, only the positive or
negative ions in the sample will be injected, neutral species will not. This can be
a significant benefit if the sample matrix is complex and the operator knows
which ionic species are of interest. A disadvantage of this technique is the non-
linear relationship between the field-strength used and the mobility of the ions in
the sample. The most mobile ions will migrate through the sample and into the
capillary faster than slower ions. There will therefore be an enhanced accumula-
tion of the most mobile ions that can lead to inaccuracies in quantitation unless
the sample matrix is well defined and stable. A good use of electromigration in-
jection is in the analysis of very low levels of anions and cations in ultra-pure
water (Fig. 14.10).
14.2.10 Detection
There are a number of detection techniques available for CE systems; all are
taken from HPLC, although a number of modifications are necessary for opti-
mum CE performance. The detector types and their uses are presented here.

416 Chapter 14
4
3
5
2
i
i ,, , , , ~ I I l ~ I ~ l ~ ~ ~
3.60 3.80 4.00 4.20 4.40 4.60 4.8

minutes
Fig. 14.10. Analysis of anions at trace level in deionized water containing 2000 ppm boron (as
boric acid). Electrolyte 9 mh4 sodium chromate, pH 7.8, 0.5 mh4 CIA-PakTM anion-BT. Detection
indirect UV at 254 nm. Injection by electromigration (45 s at 5 kV). Courtesy of Millipore-Waters.
Peak I , chloride (2 ppb); 2, sulfate (3 ppb); 3, nitrate (n.q.); 4, chlorate (30 ppb); 5, fluoride
(2 PPb).
I4.2.10.1 W detection
UV is the most widely used detection technique in CE. The detector flow-cell
is normally obtained by removing the polyimide sheath from the capillary at a
predetermined distance from the sample end. The flow cell thus created is
mounted into an appropriate optical assembly. The very short optical pathlength,
corresponding to the internal diameter of the capillary, limits the sensitivity of
the technique, although it does lead to sharp analyte peaks. Recently two variants
of flow-cell design have been introduced, both of which aim to increase the opti-
cal pathlength and increase sensitivity. The Z-Flowcell increases the pathlength
to 3 mm, while the Bubble cell increases the pathlength to 300pm. Both result
in increased sensitivity but with a decrease in resolution.
Detection wavelengths are selected either by lamp and filters (discretely vari-
able) or by a diffraction grating and polychromator lamp. Changes in absorbance
due to the analyte are detected by photodiode, or photomultiplier, or photodiode
array (Fig. 14.11). Each of these approaches has advantages. Discretely variable
detectors have the advantage of simplicity and unmatched sensitivity at specific
wavelengths. The tunable detectors offer more flexibility as they can be tuned to
the appropriate wavelength for the analyte, but they often lack sensitivity at ul-
tra-low wavelengths (lower than 190 nm). Photodiode array detectors for CE
have only recently been introduced and offer increased information about the
sample. The main benefit of CE, however, is the ability to resolve closely related
species. The benefits of photodiode array detection in CE are not as obvious as
they are for the lower resolution techniques such as liquid chromatography.
An alternative use of UV detection is termed indirect UV. The technique re-
lies on the use of an electrolyte having a strong chromophore at a specific
wavelength such that non-UV absorbing analytes can be detected. (Fig. 14.12).
A 2a
C
1 4 5 7 9
......... ... ..................
/'
....
....
,..'
B Zb
o'..
Za "..*
7 .:-.
3 .....
@
\ .......................................................
-9"
2a
0
.y
........................ ...
0 f
...
2b
Fig. 14.11. Four most common UV visible detectors in capillary electrophoresis: (A) fixed-
wavelength detection; (B) variable-wavelength detectors; ( C ) scanning monochromator detector;
(D) photodiode array detector. 1, light source; 2a and 2b, deuterium or tungsten switchable lamp
sources; 3, mirror; 4, capillary; 5, filter; 6, grating; 7, photodiode or photomultiplier; 8, diode array.
Reprinted with permission [4].

418 Chapter 14
1 CIP-Sullonate 25 ppnl
;I1I
2 CtO-Sullonnte 25 ppiii
3 C9-Sulloriale 25 ppiii
4 CB-Stillonale 25 p p i ~
5 C7-Sullonate 25 ppit~
1
6 C6-Sulloiiale 25 ppiil
7 CS-Sulfonate 25 ppiit
8 C4-Sullonale 25 ppri~
-c
JL
I 1 I I
6.0 8.0 10.0 12.0
Migralion lime (Mnules)
Fig. 14.12. Indirect UV detection of sulfonic acids. Electrolyte: naphthalene sulfonic acid and ace-
tonitrile. Capillary 75 p m X 60 em. Injection 30 s, hydrostatic. Voltage 20 kV. The naphthalene
sulfonic acid provides the UV background absorbance necessary for the detection of the non-
absorbing analmytes. This electrolyte was also chosen as its mobility is close to the mobility of the
analytes. This is necessary for symmetrical peaks [4]. Courtesy of Millipore.
The use of indirect U V detection for ion analysis by CE is described extensively

by Jandik and Bonn [ 121. This detection mode is universal and very sensitive
with electrolytes such as chromate or parahydroxybenzoate. Due to differences
in the separation mechanism, there are no system peaks in CE, unlike the use
of indirect W in HPLC. The z flow cell or bubble cell do not offer significant
advantages for indirect W, as they do not improve the signal to noise ratio.
14.2.10.2 Fluorescence, indirect fluorescence, laser-inducedfluorescence
Direct fluorescence detection allows very low detection limits for compounds
containing fluorophores, or those which can be derivatized to form fluorescent
compounds. Most applications concern peptides, amino acids and proteins. The
most sensitive detectors are those using lasers as the excitation source. The exci-
tation wavelengths are 442 and 326 nm with a helium-cadmium laser, 488 nm
with a cadmium laser or argon ion laser. It is usually necessary to derivatize the
sample to be able to use these wavelengths. Classical derivatives such as ortho-
phthaldehyde or dansyi chloride are used with normal fluorescent detection for
amino compounds, but special reagents have ben developed for use with laser
induced fluorescence detection. Using these compounds, very good detection
limits can be obtained. Ten attomoles of amino acids can be detected [25]. Deri-
vatization at such low levels is very difficult and these results were obtained by
dilution of the sample from higher (nanomole) concentrations.
Indirect laser induced fluorescence detection permits the detection of non-
fluorescent analytes by using an electrolyte containing a fluorescent compound.
Results have been published for the determination of both cations and anions
using indirect laser fluorescence. The detection limits, however, are in the same
order of magnitude as the detection limits by indirect UV with a less expensive
instrument.
14.2.10.3Amperometric detection, conductometric detection, MS detection
All the detection modes used in W L C will be available for CE in the near
future. The following three modes of detection are still in the development stage
but will certainly soon be in routine use.
Amperometric detection. This technique is often termed electrochemical detec-

tion, it requires that the analyte molecules have a redox potential, i.e. that they
can be either oxidized or reduced by applying a low voltage. Classical examples
are catecholamines, carbohydrates and some inorganic compounds such as sul-
phite, sulphide, cyanide and iodide. Detection of metal ions has also been de-
scribed. The design of a cell is shown Fig. 14.13. Working electrodes can be
made of carbon fibre (catecholamine analysis), copper (carbohydrates), silver
(sulphur species), mercury, gold or platinum. Interest in this technique lies in the
selectivity and sensitivity it can offer, for example attomoles of catecholamines
can be detected [26]. Compared to HPLC, amperometric detection in CE gives
better coulombic efficiency (percentage of the electroactive analyte that is oxi-
dized or reduced in the cell).The main difficulty to overcome in CE is that rou-
tinely the detector is set to approximately 1.0 V (in order to oxidizeheduce the
analyte) which generates a current of nanoamps, in the presence of the high volt-
age and microamp current generated by CE. It is therefore necessary to ground
Fig. 14.13. Amperometric detection cell design. Reprinted with the permission of ref. [25]. A,
fused silica capillary; B, drop of carrier electrolyte; C, stainless steel plate; RE, reference electrode;
WE, working electrode; AE, auxiliary electrode.

420 Chapter I4
the high voltage at the detection end of the capillary before the amperometric
detector flow cell [27]. A similar approach is required for conductometric detec-
tion.
Conductometric detection. This mode of detection works in exactly the same

way as the HPLC detector and it is an attractive detector to use for measuring
ions. The detection principle is to measure the change in conductance of a solu-
tion due to the presence of an ion. The conductance can either be enhanced or
reduced by the presence of the ion. It is difficult to measure small changes in
conductance in the presence of the highly conducting CE electrolyte and high
voltage. An interesting experimental approach is the use of chemical suppression
1281. Using this method, good detection limits were presented for anion analysis,
but a number of problems still need to be resolved. It is difficult to assemble the
suppressor and conductivity cell on the capillary and the lifetime of the suppres-
sor is compromised by the cationic surfactants used to control the EOF. If the
EOF is not controlled, the separation becomes very unstable. There are difficul-
ties therefore in the selection of electrolytes which are compatible with both
separation and suppressioddetection.
Once these problems have been overcome, conductometric detection of ions
separated by CE should offer advantages against present methods of detection.
Mass spectroscopy detection. The mass spectrometer is an attractive detection

technique as it can give absolute identification of the separated analytes based on
their molecular mass and, if the appropriate technique is used, mass spectrum. It
is for this reason that LC-MS is a fast growing technique. One of the difficulties
with LC-MS is the incompatibility between the optimum flow rates for LC
analysis versus the optimum flow rates for most MS interfaces. One possible
solution attempted by MS manufacturers is to modify the interface to accommo-
date high flow rates. This can, however, compromise the performance of the MS.
An alternative solution is to use capillary LC with microbore columns and flow
rates in the microliter per minute range. In CE, the EOF usually ranges from 0.1
to 1 ,uI/min, which makes it more suitable for MS interfaces than HPLC.
A review of CE-MS published recently [28] comments that the electrospray
ionization interface (ESI) is more frequently used than fast atom bombardment
(FAB). This is probably due to the extensive use of ESI for protein and peptide
analysis, for which it is well suited, as is CE.
There are compatibility problems with CE electrolytes and MS, this can be
reduced by the use of a sheathing liquid which effectively dilutes the CE electro-
lyte to ionic levels that the MS can tolerate (Fig. 14.14). The combination of the
resolving capacities of MS and CE are synergistic. The only issue is sensitivity
since the injected amounts in CE are so small, and the need to dilute the CE
Capillary electrophoresis in the petroleum industry 42 1
Fund sllka ESI buff- flow Hoc

E 5 rotcogr,
~elate
-r
t+lOpl/mln) lo,Coowr
'Y
Inol I Purp II
Nz
Fig. 14.14.CEMS interfacing. Reprinted with the permission of ref. [26].
electrolyte. Reducing the elution speed (by controlling the CE voltage) and by
using optimized injection techniques (sample stacking, isotachorphoresis pre-
concentration), it is possible to improve detection limits. Selected ion monitoring
(SIM) also brings an additional gain in sensitivity.
14.3 APPLICATIONS OF CAPILLARY ELECTROPHORESIS
Capillary electrophoresis can be applied to most of the areas previously asso-

ciated with liquid chromatography. The following review is not intended to be
exhaustive and new applications are being published almost daily.
The majority of applications published to date relate to the pharmaceutical in-
dustry. Most new pharmaceutical compounds are now subjected to various
modes of CE analysis during their development stages and as these become for-
mulated into new drug products, it is apparent that CE will play an increasing
role in this field. An area of particular importance is the use of CE to provide
elegant separation of optical isomers in the presence of electrolyte additives such
as cyclodextrin (Fig. 14.15). Most pharmaceutical drugs are cationic and are as-
sociated with a counter-ion such as sulphate or chloride. The quantitation of the
counter-ion and of possible anionic impurities is readily achieved by capillary
ion analysis (CIA) which allows rapid and accurate mass balance problems to be
resolved (Fig. 14.16).
Within the chemical industry in general, the most common application of CE
is the analysis of mixtures of small ionic species where it is used for process
monitoring, quality control product formulation and environmental monitoring
422 Chapter 14
f I I I 1
0 S 10 1s 20
n
m o (mfn)
Fig. 14.15. Separation of enantiomeric compounds. Reprinted with the permission of ref. [27]. I,
(-)-Pseudoephedrine; 11, (+)-ephedrine; 111, (-)-ephedrine; IV, (+)-pseudoephedrine. Electrophero-
gram which shows the separation of the enantiomers and diastereomers of ephedrine and pseu-
doephedrine. The separation was achieved with the 25 p m capillary and I .5 mM p-CD-SBE(1V) in
20 mM Tridphosphate buffer (pH 2.5). The field strength was 210 V cm-* and sample concentra-
tions were 0.4 mM. Note the two adjacent chiral centres and the baseline separation of all four
isomers.
(including waste water, smoke emissions, working atmospheres and biological

fluid monitoring). The ease of use and low running costs make CIA attractive for
the determination of ions in chemicaHpetroleum plants. Applications range from
deionized water to waste water [30]. Figure 14.17 shows the analysis of waste
water from a polyethylene plant. The separation of organic acids is also an im-
portant application where CIA offers fast analysis with little risk of error from
interfering compounds. The determination of surfactants and impurities in sur-
factants is still difficult by HPLC or IC. CIA has been shown to offer an excel-
lent solution to this problem [3 11.
Monitoring manufacturing processes, mainly by CIA is now well accepted in
the pulp and paper industry [32] and in the atuminium industry [33]. Organic
acid impurities in the manufacture and processing of terephthalic acid are now
widely monitored by CIA. The analysis of agricultural fertilizers, especially the
analysis of nitrate and nitrite is being developed. This has revealed a further use
for CE in forensic analysis. Both inorganic (nitrate and chlorate [34]) and or-
ganic compounds [35] are present in explosive residues and can be detected fol-
lowing acts of terrorism. The forensic determination of drugs of abuse has also
been published [36].
The determination of ions by CIA for environmental control is well estab-
lished in many areas. Samples range from tap water to surface water, rain, air
and waste water. Inorganic anions and cations and organic acids are readily
1
I I I I
0.W im 200 3w 4m
Retention time in minutes
Fig. 14.16. The analysis of a pharmaceutical formulation by capillary ion analysis. Note the pres-
ence of sulphate as a contaminant, the presence of sulphate caused discrepancies in the mass-
balance calculations for this formulation.
6 i
1
I " ' I " ' l " ' l " ' l " ' l " ' 1 '
2.60 2.80 3.00 3.20 3.40 3.60 3.80
minutes
Fig. 14.17. Capillary ion analysis of a water sample from a waste water treatment plant.. Analytical
conditions: 5 mM sodium chromate, 0.5 mM CIA-Pakm anion-BT. Hydrostatic injection for 30 s.
Indirect UV detection at 254 nm. Peak 1, chloride; peak 2, sulfate; peak 3, nitrite; peak 4, nitrate;
peak 5, organic acid; peak 6, bicarbonate. Courtesy of Millipore.

424 Chapter 14
analysed. Organically bound metals require extensive sample preparation proto-

cols before final analysis by CIA. The results obtained by CIA in these applica-
tions are in good agreement with those obtained by other methods and CE is
faster and more flexible than IC and can frequently replace gradient IC.
14.4 CONCLUSION
The interest in CE and the rapid evolution and acceptance of the technique
suggests that the market for CE instrumentation will probably grow to the size of
the HPLC and GC markets. It is important to remember that CE should be con-
sidered as a complementary technique to HPLC, IC and GC and not as a rival.
The main advantages of CE include high resolution, speed of analysis and sensi-
tivity when expressed as minimum detectable mass. It can also be considered to
be a user-friendly technique due to the absence of a packed column which is of-
ten a source of problems in HPLC and IC. Another important aspect is that the
low sample volumes required means that CE can almost be considered as a non-
destructive method of analysis. The low consumption of solvents, buffers and
gases reduces the environmental burden .As in any new method, there have been
impressively rapid developments in instrumentation and applications.
The small diameter of the CE capillary can be considered a limitation of the
technique in that the sample size is very small. In terms of minimum detectable
concentration, HPLC has a lower limit of detection. A further limitation is the
inability of the capillary to accommodate widely different analyte ion concentra-
tions compared to IC and HPLC. The linear range of the detector can be limited
to only three orders of magnitude but improvements in technology are taking
place. The low sample capacity of CE may be a disadvantage for those interested
in collecting the separated fractions even when using optimized devices such as
a membrane fraction collector [37], even so the size of the collected fractions
remains very small.
The electroosmotic flow can also be considered to be a limitation although the
main advantages of CE are due to the EOF. The EOF permits short migration
times, migration of non-ionic substances and the separation of both anionic and
cationic species simultaneously. It is difficult, however, to maintain a constant
EOF and therefore reproducible migration times. EOf fluctuations due to tem-
perature changes or to electrolyte exhaustion have been solved by the manufac-
turers but injected samples may modify the capillary wall and cause a change in
the EOF. Coating the capillaries, the addition of a zwitterion or cationic surfac-
tant, or purging the capillary between injections all help to stabilize the EOF. In
FZCE or MEKC, the most reliable method is to thoroughly rinse the capillary
with extensive base washes before use followed by frequent rinses between
analyses. For compounds insoluble in water, CE is not always the best method to
use.
CE is a new technique; if it follows the same development and growth pattern
as GC and HPLC we can expect to see many improvements in the near future.
The 5th International Symposium on High Performance Capillary Electrophore-
sis (Orlando, 1993) presented a good overview of the developments in CE. These
were concerned not only with those techniques covered here but also with many
others. It is now possible, for example, to reduce analysis time down to a few
seconds or too carry out hydrolysis or derivatization reactions in the capillary
prior to separation.
Instrument improvements will include the control of EOF using an external
radiofrequency field, sophisticated control of the applied voltage for better re-
producibility and improved MS detection. Interesting hrther developments in-
clude the miniaturization of the CE system and its auxiliaries by etching onto a
silica chip. It may be possible in this way to reduce the overall size to that of an
electronic component and to be able to use it as an in-vivo sensor [38].
14.5 REFERENCES
1 A. Lodge, B.A. Thesis (1886).

2 S. Compton and R. Brownlee, Biotechniques, 6 (1988)432.
3 A.Tiselius, S. Hjerten and S. Jerstedt; Arch. Ges. Virusforsch., 17(1965) 512.
4 S. Hjerten, Chromatogr. Rev., 9 (1967)122.
5 R. Virtenen, Acta Polytech. Scand, 12 (1974)I.
6 F.E.P. Mikkers, F.M. Everaerts and T.P.E.M. Verheggen, Anal. Chem., 169 (1 979)11.
7 J.W. Jargenson and K.D. Lukacs, Anal. Chem. 53 (1981)1298.
8 S. Terabe, K. Otsuka, K. Ichikawa, A. Tsuchiya and T. Ando, Anal. Chem., 56 (1984)111.
9 A. Cohen and B.L. Karger, J. Chromatogr. 397 (1987)409.
10 B.B. Van Orman and G.L. McIntire, J. Microcolumn Separation, 6 (1989)289.
11 S.F.Y. Li, Capillary Electrtophoresis, Journal of Chromatography Library, Vol. 52, Elsevier,
Amsterdam (1992).
12 P. Jandik and G. Bonn, Capillary Electrophoresis of Small Molecules and Ions, VCH, New
York (1993).
13 J. Vindegol and P. Sandra, Introduction to Micellar Electrokinetic Chromatography, Huthig,
Heidelbert (1992).
14 T. Tsuda, J. Liquid Chromatogr. 12 (1989)2501.
15 M. Merion, B. Bell-Alden, E. Grover, U. Neue and J. Peterson, poster presented at HPCE
(1991).
16 K. Otsuka, S. Terabe and J. Ando, J. Chromatogr., 332 (1985)219.
17 K. Otsuka, S. Terabe and J. Ando, J. Chromatogr., 396 (1987)350.
18 K.Otsuka and S. Terabe, J. Chromatogr. ??? (1989) 150.
19 S. Terabe, K.Otsuka and T. Ando, Anal. Chem., 61 (1989)251.
20 S. Terabe, Y. Ishihaua, H. Nishi, T. Fukuyana and K. Otsuka, J. Chromatogr. 545 (1991)359.
21 H.Nishi, T. Fukuyana, M. Matsuo and S. Terabe, J. Microcolumn Separation l(5) (1989)234.
426 Chapter 14
22 J.R. Mazzeo, J.A. Martineau and I.S. Krull, Methods, 4(3) (1992) 1.
23 J. Liu, Y.Z. Hsieh, D. Wiesler and M. Notovny, Anal. Chem., 63 (1991) 408.
24 AS. Cohen and B.L. Karger, J. Chromatogr., 397 (1987) 409.
25 R.W. Wallinford and A.G. Ewing, Anal Chem., 61 (1989) 229A.
26 R.W. Wallinford and A.G. Ewing, Anal Chem., 60 (1988) 258.
27 P.K. Dasgupta and L. Bao, Anal. Chem., 65 (1993) 1003.
28 R.D. Smith, J.H. Wahl, D.R. Goodlett and S.A. Hofstadler, Anal. Chem., 65( 13) ( 1993) 574.
29 C.S. Weiss, J.S. Hazlett, M.H. D a m and M.H. Danzer, J. Chromatogr., 608 (1992) 325.
30 G. Bondoux, P. Jandik and W.R. Jones, J. Chromatogr., 602 (1992) 79.
31 Y.M. Liu and S.J. Sheu, J. Chromatogr., 637 (1993) 219.
32 D.R. Salomon and J. Romano, J. Chromatogr., 602 (1992) 219.
33 S.C. Grocott, L.P. Jefferies, T. Bowser, J. Camevale, P.E. Jackson and B.F. Kenny, J. Chro-
matogr., 602 (1992) 257.
34 T. Kishi, J. Nakamura, Y. Komo-oka and H. Fukuda, 4th Int. Symp. on Analysis and Detec-
tion of Explosives, Jerusalem (1992).
35 W. Kleibohmer, K. Camman, J. Robert and E. Mussenbrock, J. Chromatogr., 638 (1993) 349.
36
37 F. Tagliaro, C. Poiesi, R. Aiello, R. Dorizzi, S. Ghielmi and M. Marigo, J. Chromatogr., 638
(1993) 303.
38 Y.F. Cheng, M. Fuchs, D. Andrews and W. Carson, J. Chromatogr., 608 (1992) 109.
427
Subject Index
additives analysis by HPLC, 366-368 - capillary tubes for, 412-413

amperometric and conductometricdetectors - applications, 421-424
for CE, 419-420 - detection methods, 415-42 1
associated gas analysis by temperature pro- - development of, 401404
gramming, 18 - free zone 404-408
associated gas - future prospects, 424-425
- higher boiling compounds, 16-21 - gel filled 411-412
- two detectors in series, 16 - injection methods, 414-415
ASTM - power supplies, 4 13
- D1160,41 - techniques and apparatus, 404-421
- D2001, D2002, D2003, D2549 and D3712, - temperature control, 413-414
371 capillary ion analysis (CIA), 421-424
- D2892,41 capillary isoelectric focussing, 412
- D3710 for gasolines, 46 carrier gases for high temperature GC, 71-
- D86,41 72
atomic emission detectors (AED), 160-200 coal and shale, 131
applications of, 191-197 column switching
excitation sources for, 161-162 - as a sampling technique, 258-263
historical development of, 162-164 - modes for multidimensional GC, 235
home-built, 164-1 84 - techniques, 241-250
linear dynamic range of, 177-1 80 coupling a capillary pre-column to a capillary
microwave cavities, 164-168 column, 260-262
microwave power supply for, 166 coupling a packed pre-column to a capillary
operating limits, 170-180 column, 262-263
scavenger gas for, 176-177 crude oil, sulfur compounds with an SCD,
schematic diagram of system, 165 224-227
selectivity of, 180-1 83 cutting techniques, 245-249
spectrometer for, 167-169
upper limit of detection, 174-1 76 data handling methods, 375-399
Deans switching of multi-column systems,
backflushing 253-255
- in gas analysis, 10, 13-16, deconvolution of partially resolved peaks,
- summation of backflushed compounds, 382-3 84
249-250 detector linearity in quantitation, 380
- techniques, 244-245 detector signal processing, 380-384
baseline assignment in quantitation, 382 dielectric constant detector for HPLC, 360-
biomarkers in crude oil, 137 361
diesel fuel
calibration curves for quantitative analysis, - hydrocarbon group analysis by SFC, 336-
392-393 343
capillary electrophoresis (CE), 401-426 - sulfur compounds with an SCD, 221-224
428 Subject Index
diode array detectors for HPLC, 354-355 - future trends, 372

electro-osmotic flow, 405-407 - history of application to petroleum analysis,
electrochemical detectors for HPLC, 355-356 347-348
electronic filtering of detector signals, 381 - pumps, 348-350
errors in quantitation - sample injectors, 350-351
- due to baseline assignment, 390 - table of applications in petroleum analysis,
- due to peak asymmetry, 386-387 363-364
- due to peak overlap, 386 - with mass spectrometry, 356-357
- transmission of systematic errors, 387-389 - with NMR and IR, 357-358
- from noise, 385 hydrocarbon gases

errors in quantitative results, 384-390 - calibration mixtures for, 38-39
- quantitative measurement, 25-28
FIA analysis of gasoline (ASTM D 1319, - rapid analysis, 21
IPl56), 370 - sample handling, 37-38
hydrocarbon group analysis by SFC, 328-329
gas analysis hydrocarbon type analysis by HPLC, 368-
- detector response factors, 27-28 370
- precision, 26-27 hydrodynamic chromatography (HDC), 95-96
gas samples, air contamination, 25 HDC
gasoline - block polymer analysis, 106-109
- anti-knock additives in, 143 - C O ~ U for,
~ ~ 103-104
S
- composition, trends in, 144-146 - detectors for, 102-103
- emissions from, I44 - effect of column coiling, 103-104, 117-
- hydrocarbon group analysis by SFC, 330- 119
334 - instrumentation for, 100-104
-- oxygenated compounds in, 143, 146-147 - metal detergent sizing, 109-1 11
- oxygenates with an atomic emission detec- - separation of polystyrene standards, 104-
tor. 197 105
- sulfur compounds with an SCD, 218-220 - tubular pinch in capillaries, 11 1-1 17
geochemistry, high temperature GC for study FIDC
of, 127 - in capillaries, 11 1-1 17
- in microcapillaries, theory of, 96-99
heterocyclic nitrogen and sulphur compounds HDC applications
by HPLC, 365-366 - in capillaries, I 19-1 20
Hewlett Packard AED - in microcapillaries, 104-1 1 1
- 5921A, 184-191 - in packed columns, 120-124
- columns for, 186
- computer controls for, 190 injector design, effect on quantitation, 378
- photodiode array spectrometer for, 189- integrator calibration, 390-393
190 integrators
-- plasma cavity for, 187-1 89 - accuracy and precision, 384-385
- sensitivity and selectivity, 190-191 - role of, 376-378
fiPLC
- analysis of petroleum mixtures, 347-374 kerogen
- apparatus for petroleum analysis, 348-361 - and other macromolecules in crude oil,
- columns, 351-353 128-130
- evaporative light scattering detectors for, - pyrolysis-GC-MS of, 129
3 5 9-3 60 kerosine and naphtha hydrocarbon group
- FIDfor. 356 analysis by SFC, 334-336
Subject Index 429
light scattering detector, 86-87 PCBs by HPLC, GPC and GC, 371
limitations of the detector signal in data han- peak area measurement, 381-382
dling, 378-380 peak asymmetry ratio, 377
live column switching, 255-258 petroleum geochemistry, 130-13 1
- using GC, 133-135
macromolecules, size by HDC, 98-100 - using GC-isotope ratio MS, 137
mass spectrometric detection for CE, 420-421 - using GC-MS, 135-137
metal porphyrins in crude oil with an AED, - using LC and TLC, 131-133
197 - using LC-MS, 137-138
methyl tert-butyl ether (MTBE), 144 - using SEC and SFC, 138
micellar electrokinetic chromatography pneumatic switching of multi-column systems,
(MEKC), 408-4 1 1 252-253
microwave cavities for atomic emission detec- polyacrylamides by hydrodynamic chromatog-
tors, 164-166 raphy, 121
multi-column systems, 23 1-268 polycyclic aromatic hydrocarbons by HPLC,
- analysis time reduction, 232 362-3 65
- extended column life, 233
- for natural gas, 11-16, 19-22 quantitation
- improved detection limits, 233 - by area normalisation, 391
- increase in resolution, 232 - by internal and external standards, 391-392
- instrumentation for, 250-258 - by peak heighupeak area, 390-391
- strategies for application of, 263-266 - in HPLC, 361-362
multi-dimensional GC, definition of, 233-234
recycle Chromatography,250
natural gas refinery gases, 28-38
- analytical procedures for, 10-28 - adsorption columns for analysis of, 31-37
- backflushing higher boiling components, - sulfur compounds with SCD, 216-218
12-13 retention of material on columns, effect on
- calorific values, 4, 5-7 quantitation, 379
- compression factor, 4 RI detectors for HPLC, 358-359
- dewpoint, 4, 8-10,
- genera1 properties, 2 4 selectivity tuning of serial coupled columns,
- isothermal analysis, 10-1 5 238-24 1
- relative density, 4 separation performance of columns (no. of
nitrated PAHs by HPLC, 365 possible peaks), 23 1
noble gases in natural gas with an AED, 197 serial coupled columns, 235-238
SFE-GC, 291-299
0-FID detector, 143, 147-152 - with AED, 296-297
0-FID - with FTIR-MS, 298-299
- applications other than gasoline analysis, - with MS, 294-296
156-157 SFE-HPLC, 299-300
- for total oxygen determination, 154-155 - withMS,300
- quantitative analysis of oxygenated com- signal smoothing algorithms, 38 1
pounds, 152-155 signal to noise ratio in quantitation, 379-380
- response factors for various oxygenated silica particles by hydrodynamic chromatogra-
compounds, 153 phy, 122
- selectivity of, 155-156 simulated distillation
optimisation of quantitative analysis, 395-397 - ASTM Method D2887,42
430 Subject Index
- hy SFC, 322-327 - of liquids, 281-283

- capillary columns for, 44 - off-line applications, 284-290
- definition of, 4 1 - off-line, 283-284
- for coal liquids, 45 - on-line applications, 291-301
- for crude oils, 50-51 - on-line with an analytical technique, 284
- for gasolines, 46-47 - under dynamic conditions, 280-281
- for used engine oils, 51-52 - under static conditions, 280
- lubricating oils and residues, 47-50
- multi-element, 45 tangenVperpendicu1arpeak splitting in inte-

- precision of ASTM D2887,42-44 gration, 390
- process control, 51-52 two detectors in series, 10, 16
- SFC for, 48-49 type analysis of crude oil fractions, 131
- with an AED, 191-197
stopped flow techniques, 250 Unresolved complex mixture (UCM), 134
sulfur chemiluminescence detector (SCD), UV detectors for CE, 4 16.41 8
201-229 UV fluorescence detectors
- applications, 216-227 - for CE, 418-419
- chemiluminescence cell, 204 - for HPLC, 355
- columns for, 2 14-2 15 UV/visible detectors for HPLC, 353-355
- flame source for, 204
- flameless version, 2 12-2 14 validation of quantitative analysis, 393-395
- principle of operation, 203-204 valve switching of multi-column systems, 25 1-
- relative response factors, 208 252
-- schematic diagram of, 205 viscosity detector for wax analysis, 88
- sensitivity and selectivity, 209-212
sulfur-selective detectors for GC, summary of wax analysis, 55-93
currently available, 202-203 waxes
supercritical fluid - by IP method 372185, 57
- definition of, 269 - carbon number distribution, 75-78
- properties of, 269-270 - detectors for, 64-65
supercritical fluid chromatography (SFC), - general properties of, 55-57
305-3 45 - high temperature columns for, 65-70
- applications, 322-343 - programmable temperature vaporizer
- column ovens, 309 (PTV), 61-64
- detectors for, 3 10-322 - quantitative analysis, 70-75
- history of the technique, 305-306 - sample introduction, 6 M 4
- instrumentation, 306-322 - SFC Of, 78-81
- pumps for mobile phase, 307-309 - size exclusion Chromatography of, 81-89
- sample introduction, 309-3 10 Wobbe Index of natural gas, 4
- with supercritical fluid extraction, history
of, 270-27 1 Xanthane polysaccharides by hydrodynamic

supercritical fluid extraction (SFE) chromatography, 121
- advantages of, 271-275
- after derivatisation, 283 Zinc dialkyl dithiophosphates by HPLC, 371
-- equipment for, 275-280
431
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JOURNAL OF CHROMATOGRAPHY LIBRARY- volume 56

Library of Congress Cataloging-in-Publication Data

Chromatography in the petroleum industry / edited b y E.R. Adlard.

01995 Elsevier Science B.V. All rights reserved.

This book is printed on acid-free paper.

Printed i n The Netherlands

Chapter 1 . The analysis of hydrocarbon gases .................................................. 1

Chapter 2 .Advances in simulated distillation ................................................... 41

2.2.3 Aromatics and heteroatoms ................................................................. 44

Chapter 3 . The chromatographic analysis of refined and synthetic waxes ..... 55

Chapter 4 .Hydrodynamic chromatography of polymers ................................. 95

Chapter 5 . Chromatography in petroleum geochemistry ................................. 127

Chapter 7 . Microwave plasma detectors ............................................................ 159

Plasma start-up ....................................................................................

Chapter 8. The sulfur chemiluminescence detector........................................... 201

Refinery gases .....................................................................................

Chapter 9. Multi-column systems in gas chromatography ............................... 231

10.2.1 Technical advantages .. .................................................. 27 1

Chapter 11. Supercritical fluid chromatography ................................................ 305

11.2.4.3 Element specific detection.................................................. 318

Chapter 13. Modern data handling methods ....................................................... 375

13.7.1 Meaning of validation ......................................................................... 393

Subject Index ............................................................................................................ 427

Although the spectacular development of gas chromatography in the 1950s

D.J. ABBOTT Esso Research Centre, Analytical Group, Abingdon, Oxford-

1. ROBERTS Analytical & Applied Science Division, BP Research & Engi-

The analysis of hydrocarbon gases

Hydrocarbon gases can be categorized in a number of ways, one of which is to

by means of valves. This complexity, which is more daunting in prospect than in

1.2 NATURAL GAS

Hydrocarbon gases arise naturally from a variety of sources. Bacterial fermen-

Natural gas is part of a continuum of hydrocarbons, ranging from methane to

For identification of source, the concentrations of the inert components and

1.2.1 Analytical requirements

N2 C02 C1 C2 C3 164 nC4 nee= IC5 n-C5 C6+

Fig. 1.4. Calorific value errors.

defined, and so must its contribution to CV or other properties. In fact, the CV of

Fig. 1.5. Dewpoint temperature errors.

bution taken up by that of methane. The failure to measure individual compo-

be and frequently is used for calculation of several physical properties, which

1.2.2 Analytical procedures

Fig. 1.9. Three column analyser. Valve 1, position 2; valve 2, position 1

0 min 10 min 20 min

Sampla Column 3 Column 4

The above chromatograms were generated, as is evident from the conditions,

1.2.2.2 Two detectors

I . 2.2.4 Temperacure programming

Sample Molecular sieve

Fig. i 16. Combined system.

in which they have been undergoing separation. This is an over-simplification. In

0 min 4 min 8 hin

of 5-1 5 rnin. Laboratory analysers, which usually provide more information,

Fig. 1.19. Rapid analysis, light gases.

Fig. 1.20. Rapid analysis, C3 to C5 gases

Fig. 1.2 1. Rapid analysis, analysis to C8.

1.2.3 Quantitative measurement

or may not be recognized. Wherever possible, samples subject to air contamina-

1.3 REFINERY GAS

1.3.1 Analytical requirements

1.3.2 Analytical procedures

Component Symbol Boiling point ("C)

Fig. 1.25. Light gases analyser. Valve 1, position 2; valve 2, position I

Fig. 1.27. Rackflush of heavier components

Detector 1 Detector 2 cz]