Documentos de Académico
Documentos de Profesional
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45
3.
4.
5.
6.
Introduction
Why spectroscopic techniques
2.1. Analytical characteristics
2.2. Spectroscopic characteristics
2.3. Instrumental characteristics
Some theoretical elements
3.1. Classical theory
3.2. Quantum theory
3.3. Absorption and scattering: band position and intensity
Selecting the information
Spectroscopic instrumentation
5.1. Near-infrared instrument
5.1.1. Classical instrument
5.1.2. Microscope and camera
5.1.3. Sample presentation
5.2. Mid-infrared instrument
5.2.1. Microscope and camera
5.2.2. Sample presentation
5.3. Raman instrument
5.3.1. Microscope and camera
5.3.2. Sample presentation
Applications
6.1. Quantitative analysis
6.1.1. Determination of fat content
6.1.2. Determination of major and minor components
in lipid
6.2. Qualitative analysis
References
RESUMEN
Espectroscopa: Desarrollo en instrumentacin y anlisis.
Este artculo de revisin presenta las caractersticas, ventajas, lmites y potencial de tres tcnicas espectroscpicas: las espectroscopias del infrarrojo cercano, del medio infrarrojo y
Raman. Se presentan, y discuten brevemente, los aspectos tericos relacionados con estas tcnicas, la informacin que pueden
suministrar, y las principales caractersticas de la instrumentacin. La ltima parte de la revisin esta dedicada a las aplicaciones de la espectroscopia en anlisis de alimentos, con especial
nfasis en anlisis de lpidos. Las ilustraciones y los ejemplos se
han elegido para demostrar la importancia de las tcnicas espectroscpicas en los procesos en-lnea y en los laboratorios en el
anlisis de componentes mayoritarios y minoritarios.
PALABRAS-CLAVE: Anlisis - Autentificacin Espectroscopia infrarrojo cercano- Espectroscopia infrarrojo medio - Espectroscopia Raman Microscopio Quimiometria.
SUMMARY
Spectroscopy: Developments in instrumentation and
analysis.
This review presents the characteristics, advantages, limits
and potential of three spectroscopic techniques: near-infrared
KEY-WORDS: Analysis - Authentication Chemometrics - Midinfrared spectroscopy - Near-infrared spectroscopy - Raman spectroscopy.
1. INTRODUCTION
In recent years, spectroscopic techniques have
come to be regarded as attractive and promising
analytical tools for analyses conducted in research,
control or industrial laboratories. These techniques
are increasingly considered by analysts as an
obvious solution. This trend stems from instrumental
developments, the extensive use of computers and the
development of appropriate chemometric procedures.
Daily, new applications of spectroscopic techniques in
the fields of chemistry, drugs, the agro-food sector, life
sciences and environmental analysis are being
demonstrated and published. The objective should be
not only to extend the field of application of
spectroscopic techniques, but also to use them as a
matter of course in control and industrial
laboratories and to develop (according to
internationally accepted guidelines) fully validated
spectroscopic methods.
This paper presents the characteristics, advantages,
limitations and potential of three spectroscopic
techniques: near-infrared spectroscopy (NIR), mid-infrared
spectroscopy (MIR) and Raman spectroscopy.
Special emphasis is placed on the application of
these techniques to lipid analysis. The illustrations
and examples have been chosen to demonstrate the
importance of infrared (NIR and MIR) and Raman
spectroscopies both in process (on-line) control and
in laboratories for the analysis of major or minor
compounds. There are many books and papers on
the subject; most of the references included here
have been selected for their relevance and clarity.
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Table I
Analytical, spectroscopic and instrumental characteristics of NIR, MIR and Raman spectroscopies
INSTRUMENTAL
SPECTROSCOPIC
ANALYTICAL
FEATURES
NIR
MIR
Raman
/
/
-
.
.
.
.
.
-
/
/
-
na
na
na
na
Rapid analysis
Simple analysis: concentration vs peak area
Qualitative and quantitative analysis
None or reduced sample preparation
Standard calibration and transfert
procedures
Validated methods (IUPAC, AOCS, ISO)
Sample form and sample size
No use of pollutant solvents
Economic method
(e.g.
saving of labour
reduction of samples to analyse)
Direct, non-invasive and non destructive
In-situ analysis
z
z
Emission process
Intensity proportional to concentration
W ell-resolved spectra
High resolution spectra
Precise spectral frequency measurement
Non-polar group information
More significant information
Structural selectivity
Signal intensity
Trace element determination
Fluorescence-free spectra
Spectra insensitive to temperature
Sample presentation technique
Push-button instrumentation
W orks well with aqueous samples
Suitable for use at-, on- and in-line process
Compatible with suitable fibre optics
Process monitoring
Microscope
Camera (Multispectral measurement)
/
/
.
/
/
na
/
-
/
.
na
/
.
/
.
-
.
-
/
.
.
.
.
.
/
REFERENCES
(Gerrard 1991)
(Pelletier & Davis 1996)
(Baranska 1987)
(Gerrard 1991)
(Gerrard 1991)
(Hendra 1997)
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Eq1
49
Energy
a)
Overtone and
combination vibrations
Microwave Radio
1st excited
electronic state
Ground
electronic state
b)
Fundamental vibrations
c)
3
2
1
3
2
1
Ultraviolet
hMIR
IR
3
2
1
hNIR
VIS
h
0
h
UV
UV
Frequency
Near-infrared
Mid-infrared
d)
(MIR)
800
2500
25000
Ground
electronic state
Wavelength (nm)
Stokes
h0
h0
igh
3
2
1
3
2
1
3
2
1
Raman Stokes
400 50
4000
12500
hanti-
/ q 0
h0
Raman spectroscopy
hRayle
1st excited
electronic state
Virtual state
kes
/q 0
hSto
/ q 0
Rayleigh
Raman anti-Stokes
Wavenumber (cm-1)
Figure 1
Several regions of the electromagnetic spectrum and the related
spectroscopic techniques; the wavelength and wavenumber
limits of each region are shown.
Eq2
Eq 3
the
its
for
or
Figure 2
Energy level diagrams (Potential energy versus interatomic
distance) showing the basic transitions involved in NIR, MIR and
Raman spectroscopies. Two electronic states and three
vibrational states are shown.
Eq. 4
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1.2
cis C16:1
trans C16:1
cis C18:1
trans C18:1
0.8
Scatteing intensity
Grasas y Aceites
0.6
0.4
0.2
0
1690
1680
1670
1660
1650
-1
1640
1630
Figure 3
FT-Raman spectra of various pure fatty acid methylesters.
Scattered band observed in the vicinity of 1660 cm-1
characteristic of the C=C vibration of RCH=CHR groups.
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Figure 4
Near-infrared (inverse scale) (a), mid-infrared (b) and Raman
(c) spectra of a vegetable oil. The NIR spectrum is plotted as the
absorbance versus the frequency (nm), Raman spectrum as the
scattering intensity versus Raman frequency (cm-1) and MIR
spectrum as absorbance (Log1/R) versus frequency (cm-1). The
NIR spectrum was obtained by reflection mode while Raman
and MIR (with ATR accessory) spectra using FT instruments.
5. SPECTROSCOPIC INSTRUMENTATION
1300
1500
1700
1900
Fingeerprint region
C-C, C-O bending
trans -CH=CH- bending
b)
-CH=CHC-H Combination
-CH2,-CH3
C-H 2nd overtone
1100
-CH2,-CH3, H2O
Combination, O-H 1st overtone
NIR
c)
-CH2, -CH3
C-H stretching
-CH2, -CH3
C-H Combination
a)
Raman
MIR
2100
Wavelength (nm)
2300
2500
3100
2700
2300
1900
1500
1100
700
Wavenumber (cm-1)
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a) Transmission mode
Source
Wavelength
selector
Sample
Detector
Polychromatic radiation
NIR spectrum
b) Reflection mode
Wavelength (nm)
Detector
Source
Polychromatic radiation
Wavelength
selector
Sample
Detector
Figure 5
Schematic configuration of a dispersive NIR instrument;
(a) transmission mode and (b) reflection mode.
Classical instrument
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a) FT-NIR microscope
c) Chocolate spectrum
Polychromatic radiation
1 ,4
Source
Interferometer
1 ,3
A bs o r ba nc e ( Lo g 1 /R
1 ,2
1 ,1
Fixed miror
0 ,9
0 ,8
0 ,7
0 ,6
0 ,5
1 1 00
1 30 0
1 50 0
1 7 00
1 90 0
2 1 00
2 3 00
2 50 0
W a v e le ng t h ( n m )
litt
sp
am
Be
er
d) Hazelnut spectrum
Moved miror
MIR spectrum
1, 1
a b s o rb a n c e ( L o g 1 / R
0, 9
Sample
0, 8
0, 7
Fourier
transform
0, 6
0, 5
11 00
1300
150 0
1 700
1900
21 00
2 300
250 0
W a v e le n g t h ( n m )
Wavenumber (cm-1)
Detector
Figure 6
Perkin-Elmer FT-NIR microscope (a) and Visible image of the
cross-section of a bar of Belgian chocolate with hazelnut
pieces (b). Spectra of chocolate (c) and the hazelnut (d).
Sample presentation
Figure 7
Schematic configuration of an FT-MIR instrument.
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Sample presentation
a)
b)
c)
Figure 8
Several sampling devices used in MIR spectroscopy. a) ATR,
b) Thermostatised flow cell, c) IR-card.
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Interferometer
Fixed miror
Laser
Sample
Scattering radiation
Monochromatic radiation
li
sp
am
Be
r
tte
Moved miror
Scattering intensity
Detector
Raman spectrum
-1
Fourier
transform
Figure 9
Schematic configuration of an FT-Raman instrument.
Sample presentation
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6. APPLICATIONS
Various books and papers have reviewed the
application of near-infrared (NIR) (Baeten et al. 2000;
Riaublanc et al. 2000), mid-infrared (MIR) (Wilson
1990; van de Voort and Ismail 1991; van de Voort
1994; Wilson and Tapp, 1999; Baeten et al., 2000;
Riaublanc et al. 2000; Bertrand and Dufour 2000)
and Raman spectroscopies (Ellis et al. 1989; Li-Chan
1996; Keller et al. 1996; Baeten et al. 2000) in the
analysis of agro-food and lipids. The following
sections summarise the potential and the trends in
lipid analysis (quantitative, qualitative, process
monitoring).
6.1.1.
trans =C-H
Figure 10
Raman scattering bands observed in the vicinity of
1630-1690 cm-1. Spectra of milk anhydrous fat having a trans
content between 0.5 and 12%.
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Table II
Review of papers on the authentication of food products using infrared (NIR, MIR) and Raman
spectroscopy. (Modified from Baeten & Aparicio 2000)
Food product
Issue
Analytical
method
Cereals
Rice grains
Geographic origin
NIR
Coffee
Species
discrimination
NIR
FT-MIR
NIR
FT-MIR
Adulteration
Dairy
products
Milks
Butters
Skimmed milk powder
Adulteration
Adulteration
NIR &
FT-MIR
NIR
NIR
Fruit-based
products
Fruit juices
Geographical origin
and species
discrimination
NIR
Fruit pures
Jams
Meat
Meats
Adulteration
Fruit concentration
Fruit type
Process
discrimination
fats
FT-MIR
FT-MIR
FT-MIR
NIR
NIR
FT-MIR
Raman
NIR
Adulteration
FT-MIR
Raman
NIR
Geographic origin
NIR
Species
discrimination
Adulteration
Species
discrimination
Reference
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