EXPERIMENTAL

NOTES
Clarus

600 C GC-MS
Analysis and Identifcation of Faty
Acid Methyl Ester Composition in
Diferent Vegetable Oil (Biodiesel)
Source Using Gas Chromatography
Mass Spectrometry
Abstract
Vegetable oils have been attracted attention has a potential renewable source for the
production of an alternative for-petroleum based diesel fuel. Various products obtained
from vegetable oils have been proposed as an alternative fuel for diesel engines,
including neat vegetable oil, mixtures of vegetable oil with petroleum diesel fuel and
alcohol esters of vegetable oils. Out of which Alcohol esters of vegetable oils appear to
be the most promising alternative. Vegetable oils are triglycerides (glycerin esters) of fatty
acids and alcohol esters of fatty acids have been prepared by the transesterification of
the glycerides, wherein linear, monohydroxy alcohols reacts with the vegetable oils in
the presence of catalyst to produce alcohol esters of vegetable oil. The alcohol esters of
vegetable oil when used as an alternative diesel fuel have been identified as a biodiesel.
This experimental note demonstrates the analytical capability of PerkinElmer Clarus
600C GC-MS for the analysis and identification of fatty acid methyl ester composition
(Biodiesel) in various vegetable oils.
Author
Dileep Kumar Mamidala
Application Specialist Chromatography
Centre of Excellence for Analytical Sciences
PerkinElmer (India) Pvt. Ltd.
Hyderabad 500 081 India.
Email: application.india @perkinelmer.com
GC-PETRO-002
Preparation of biodiesel
Transesterification:
Weighed about 250 g of Kusum oil, Palash oil, Mehwa oil and
Thumba oil and transferred in to individual 100 mL conical flasks.
Weigh about 2.5 g of Potassium Hydroxide (1% by weight of
the oil) and dissolve it in 72 g anhydrous methanol. Completely
dissolve the KOH in methanol, apply heat if required. transfer
the resulting solution slowly to the sample in a conical flask with
stirring. After complete transfer of KOH solution in methanol
continue the stirring for 120 minutes to complete the reaction.
After completion of the reaction time pipette out and transfer
approximately 8-10 mL of reaction mixture to 15mL test tube
and keep it for phase separation.
Phase separation and washing:
After 120 minutes of reaction time, the reaction was stopped and
reaction mixture was allowed to stand overnight while the phase
separations occurred in different reaction mixtures in individual
flasks. The ester phase was decanted from the mixture and
transferred to glass column for further washing to remove the any
traces of methanolic KOH solution.
Excess alcohol and residual catalyst were washed from the ester
with water. The ester phase was placed in a glass column 1.26 cm
in diameter and 100 cm in length. Water was sprayed into the top
of the column at a low velocity. The excess alcohol and catalyst
were removed by the water as it percolated through the column.
During the washing, some of the ester formed an emulsion with
the water; a time of 24-48 hours was required for the water
phase containing alcohol, catalyst, and emulsified ester to settle
and the ester phase to become clear.
Sample preparation and analysis:
Weigh 25 mg of the ester phases of different oils and dissolve
it in 0.5 mL of n-heptane. The resulting sample solution is filled
in GC auto sampler vial and injected in GC/MS to analyze and
identify the fatty acid methyl ester composition in different types
of vegetable oils.
Instrumental Conditions:
Clarus 680 GC Optimized Experimental Conditions
Column Elite-5MS, 30m x 0.25mm x 0.25um
Injector temperature 220 C
Carrier gas Helium
Carrier gas ow 0.2 mL/min
Split ratio 50:1
Oven Programme 35 C hold for 10 min
10 C/min 200 C hold for 10 min
Total run time 36.5 min
Diluent n-Heptane
Injection volume 0.1 ul
Clarus 600 C MS Parameters
Ionization source EI
Electron energy 70 eV
Source temperature 200 C
Transfer line temp 200 C
Multiplier (V) 366 V
Ion energy 1.5 V
Scan range 10-600 m/z
Analytical results & discussion
Biodiesel samples were analyzed using PerkinElmer Clarus 600
C GC-MS in total scan mode to identify the fatty acid methyl ester
composition in different vegetable oil sources with the help of
NIST library. The samples were analyzed and the composition of
fatty acid methyl esters was identified were as follows.
Fig 1. Total ion chromatogram of Palash
biodiesel. 1. Caprylic acid methyl
ester, 2. Myristic acid, methyl ester,
3. Palmitoleic acid, methyl ester, 4.
Palmitic acid, methyl ester, 5. Margaric
acid, methyl ester, 6. Linoleic acid,
methyl ester, 7. Oleic acid, methyl
ester, 8. Stearic acid, methyl ester,
9. 11-Eicosenoic acid, methyl ester,
10. Arachidic acid, methyl ester, 11.
Heneicosanoic acid, methyl ester, 12.
13-Docosenoic acid, methyl ester,
13. Behenic acid, methyl ester, 14.
Tricosanoic acid, methyl ester, 15.
Lignoseric acid, methyl ester, 16.
Pentacosanoic acid, methyl ester, 17.
Cerotic acid, methyl ester.
Fig 3. Total ion chromatogram of Kusum
biodiesel. 1. Palmitoleic acid, methyl
ester, 2. Palmitic acid, methyl ester, 3.
Linoleic acid, methyl ester, 4. Oleic acid,
methyl ester, 5. Stearic acid, methyl
ester, 6. 11-Eicosenoic acid, methyl
ester, 7. Cis-11-Eicosenoic acid, methyl
ester, 8. Arachidic acid, methyl ester, 9.
Heneicosanoic acid, methyl ester, 10.
13-Docosenoic acid, methyl ester, 11.
Methyl 11-docosenate, 12. Behenic acid,
methyl ester, 13. Lignoseric acid, methyl
ester.
Fig 4. Total ion chromatogram of
Tumba biodiesel. 1. Caprylic acid
methyl ester, 2.Azelaaldehydic acid,
methyl ester, 3. Myristic acid, methyl
ester, 4. Pentadecanoic acid, methyl
ester, 5. Palmitoleic acid, methyl ester, 6.
Palmitic acid, methyl ester, 7. Margaric
acid, methyl ester, 8. Linoleic acid,
methyl ester, 9. Oleic acid, methyl
ester, 10. Stearic acid, methyl ester,
11. 13-Docosenoic acid, methyl ester,
12. Behenic acid, methyl ester, 13.
Lignoseric acid, methyl ester.
Fig 2. Total ion chromatogram of
Mehwa biodiesel. 1. Caprylic acid
methyl ester, 2. Myristic acid, methyl
ester, 3. Palmitic acid, methyl ester, 4.
Margaric acid, methyl ester, 5. Linoleic
acid, methyl ester, 6. Oleic acid, methyl
ester, 7. Stearic acid, methyl ester, 8.
11-Eicosenoic acid, methyl ester, 9.
Arachidic acid, methyl ester, 10. Behenic
acid, methyl ester, 11. Lignoseric acid,
methyl ester, 12. Pentacosanoic acid,
methyl ester, 13. Cerotic acid, methyl
ester.
Conclusions
This experimental result confirms the effectiveness of the
analysis procedure and also demonstrated the capability of
PerkinElmer Clarus 600 C GC-MS to identify the fatty acid
methyl ester composition (biodiesel) in different types of
vegetable oil sources.