EXPERIMENTAL

NOTES Clarus™ 600 C GC-MS

GC-PETRO-002 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

Analysis and Identification of Fatty

Acid Methyl Ester Composition in
Different 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.

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 Analytical results & discussion
stirring. After complete transfer of KOH solution in methanol
Biodiesel samples were analyzed using PerkinElmer Clarus™ 600
continue the stirring for 120 minutes to complete the reaction.
C GC-MS in total scan mode to identify the fatty acid methyl ester
After completion of the reaction time pipette out and transfer
composition in different vegetable oil sources with the help of
approximately 8-10 mL of reaction mixture to 15mL test tube
NIST library. The samples were analyzed and the composition of
and keep it for phase separation.
fatty acid methyl esters was identified were as follows.
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. Fig 1. Total ion chromatogram of Palash Fig 2. Total ion chromatogram of
biodiesel. 1. Caprylic acid methyl Mehwa biodiesel. 1. Caprylic acid
Excess alcohol and residual catalyst were washed from the ester ester, 2. Myristic acid, methyl ester, methyl ester, 2. Myristic acid, methyl
with water. The ester phase was placed in a glass column 1.26 cm 3. Palmitoleic acid, methyl ester, 4. ester, 3. Palmitic acid, methyl ester, 4.
in diameter and 100 cm in length. Water was sprayed into the top Palmitic acid, methyl ester, 5. Margaric Margaric acid, methyl ester, 5. Linoleic
acid, methyl ester, 6. Linoleic acid, acid, methyl ester, 6. Oleic acid, methyl
of the column at a low velocity. The excess alcohol and catalyst methyl ester, 7. Oleic acid, methyl ester, 7. Stearic acid, methyl ester, 8.
were removed by the water as it percolated through the column. ester, 8. Stearic acid, methyl ester, 11-Eicosenoic acid, methyl ester, 9.
9. 11-Eicosenoic acid, methyl ester, Arachidic acid, methyl ester, 10. Behenic
During the washing, some of the ester formed an emulsion with
10. Arachidic acid, methyl ester, 11. acid, methyl ester, 11. Lignoseric acid,
the water; a time of 24-48 hours was required for the water Heneicosanoic acid, methyl ester, 12. methyl ester, 12. Pentacosanoic acid,
phase containing alcohol, catalyst, and emulsified ester to settle 13-Docosenoic acid, methyl ester, methyl ester, 13. Cerotic acid, methyl
13. Behenic acid, methyl ester, 14. ester.
and the ester phase to become clear. Tricosanoic acid, methyl ester, 15.
Lignoseric acid, methyl ester, 16.
Sample preparation and analysis: Pentacosanoic acid, methyl ester, 17.
Cerotic acid, methyl ester.
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 Fig 3. Total ion chromatogram of Kusum Fig 4. Total ion chromatogram of
Column Elite-5MS, 30m x 0.25mm x 0.25um biodiesel. 1. Palmitoleic acid, methyl Thumba biodiesel. 1. Caprylic acid
ester, 2. Palmitic acid, methyl ester, 3. methyl ester, 2.Azelaaldehydic acid,
Injector temperature 220 °C Linoleic acid, methyl ester, 4. Oleic acid, methyl ester, 3. Myristic acid, methyl
Carrier gas Helium methyl ester, 5. Stearic acid, methyl ester, 4. Pentadecanoic acid, methyl
ester, 6. 11-Eicosenoic acid, methyl ester, 5. Palmitoleic acid, methyl ester, 6.
Carrier gas flow 0.2 mL/min ester, 7. Cis-11-Eicosenoic acid, methyl Palmitic acid, methyl ester, 7. Margaric
Split ratio 50:1 ester, 8. Arachidic acid, methyl ester, 9. acid, methyl ester, 8. Linoleic acid,
Heneicosanoic acid, methyl ester, 10. methyl ester, 9. Oleic acid, methyl
Oven Programme 35 °C hold for 10 min 13-Docosenoic acid, methyl ester, 11. ester, 10. Stearic acid, methyl ester,
10 °C/min 200 °C hold for 10 min Methyl 11-docosenate, 12. Behenic acid, 11. 13-Docosenoic acid, methyl ester,
Total run time 36.5 min methyl ester, 13. Lignoseric acid, methyl 12. Behenic acid, methyl ester, 13.
ester. Lignoseric acid, methyl ester.
Diluent n-Heptane
Injection volume 0.1 ul
Conclusions
Clarus™ 600 C MS Parameters
Ionization source EI
This experimental result confirms the effectiveness of the
analysis procedure and also demonstrated the capability of
Electron energy 70 eV
PerkinElmer Clarus™ 600 C GC-MS to identify the fatty acid
Source temperature 200 °C
methyl ester composition (biodiesel) in different types of
Transfer line temp 200 °C
vegetable oil sources.
Multiplier (V) 366 V
Ion energy 1.5 V
Scan range 10-600 m/z