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Fatty Acid Methylation Kit

Methyl esterification of fatty acids is commonly done prior to gas chromatography analysis to prevent peak tailing and to increase sample volatility. However, the conventional esterification procedure requires specialized equipment and high technical skill. By using the Fatty Acid Methylation Kit that employs a new reaction technique, follows by the Fatty Acid Methyl Ester Purification Kit, fatty acid methyl esterification procedure is greatly simplified.

*Patent applicant: Kyoto Prefectural University and GEKKEIKAN


  • For the analysis of volatile free fatty acids, glycerolipids and sterol esters
  • Enables reaction at 37 °C
  • Conducts methyl esterification safely and easily
  • Detects not only long-chain, but also short-chain fatty acids

Available from Sigma-Aldrich. Click here to get price information.

Targeted Fatty Acids

  • Free fatty acids
  • Glycerolipids such as triglycerides, phospholipids and glycolipids
  • Sterol esters
  • Please note this method is not applicable for sphingolipids.


Data courtesy of GEKKEIKAN

GC-MS Analytical Condition
System: 6890 GC/597 3i MSD (Agilent)
Column: DB-WAX (60m × 0.25mm, 0.25 µm)
Column Oven: 50°C (2min) to 250°C at 10°C/min, hold 250°C(8min)
Detection: MSD, 230°C
Carrier gas: He, 1.4ml/min
Injection: 2.0 µl, split-less, 250°C

Comparison Data

The methyl esterification efficiencies between the Fatty Acid Methylation Kit and conventional method is about the same independent of fatty acid side-chains.

Comparison of methylation efficiency rate

Quantitative Analysis

To use Fatty Acid Methylation Kit and Fatty Acid Methyl Ester Purification Kit offers wide dynamic range as shown by the results from dried yeast.


Fatty acid analysis of different Yeast strains

It is important to measure the exact amount of various fatty acids in different yeast strains. By using these kits, it was determined that the cerulenin-resistant strain contains more capronic acids than the parent strain.

Data courtesy of GEKKEIKAN


Easy and safe without using HCl.

Conventional Method

Quantitative capability of the conventional method is questionable due to the high heating requirement. The high temperature causes the degradation of unstable fatty acids (polyunsaturated and cyclopropane fatty acids) and the evaporation of short-chain fatty acid alkyl esters.

Sample Preparation

Staring Material Procedure
E. coli or Yeast Centrifuge cell culture medium of E. Coli or Yeast in a centrifuging tube and then freeze-dry about 20 mg of the pellet. An alternate method for the E. coli sample is to dry in a vacuum desiccator for 1-2 hours.
Blood Apply 0.04 ml of heparin-treated blood to antioxidant agent BHT-treated filter paper and dry it in a vacuum desiccator for 30 min. or let it dry naturally for more than 2 hours. To get complete methylation, spread the heparin-treated blood on filter paper thinly. [How to make antioxidant agent BHT-treated filter paper] Soak filter paper such as Whatman 3M or ADVANTECH No. 2 into Acetone containing 0.05% BHT for several minutes and then repeat this process. (Prepare an alternative new solvent for the second immersing). Let it dry naturally at room temperature, then put it in a vacuum desiccator for 30 min. or in a desiccator overnight. The paper with a square 1.5 cm on a side is suitable for methylation application. Please note that thicker filter papers, e. g., blood collecting filter papers have low methylation efficiency
Rat Liver Lyophilization of 15 mg of rat liver. Please note that drying rat liver in a vacuum desiccator decreases methylation efficiency.
Edible Oil Less than 4 mg of edible oil is suitable for the methylation application.
Soybean Flour Less than 20 mg of soybean flour is suitable for the methylation application.
Fish Put 200 mg of fish meat, e.g., Japanese horse mackerel, into a test tube, and add 2 ml of Isolation Solution ,then mesh the fish meat with a glass rod. After vortexing, take 0.5 ml of supernatant to a new test tube, and then dry it in a rotary evaporator, vacuum desiccator, or N2 gas.

For procedure of sample preparation of Glycerolipids analysis, please refer to its instruction.

Kit Contents

Product NamePKG SizeQTY
Methylation Reagent A 50 ml 1
Methylation Reagent B 50 ml 1
Methylation Reagent C 50 ml 1
Isolation Reagent 250 ml 1


Related Products

Fatty Acid Methyl Ester Purification Kit (50 tests)

Product NamePKG SizeQTY
Conditioning Solution 200 ml 1
Washing Solution 200 ml 1
Eluting Solution 200 ml 1
SPE Cartridge Column   50 pcs


Protocol (PDF): Fatty Acid Methylation Kit


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  2. Wijaya, Hans, et al. "Concentration of Lipase from Aspergillus oryzae Expressing Fusarium heterosporum by Nanofiltration to Enhance Transesterification." Processes 8.4 (2020): 450.
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  4. Huang, Xiaochen, et al. "Fermentation of pigment-extracted microalgal residue using yeast cell-surface display: direct high-density ethanol production with competitive life cycle impacts." Green Chemistry 22.1 (2020): 153-162.
  5. Kanai, M., Yamada, T., Hayashi, M. et al. Soybean (Glycine max L.) triacylglycerol lipase GmSDP1 regulates the quality and quantity of seed oil. Scientific Reports 9, 8924 (2019) 
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Ordering Information

Product Cat.No. Price(US$)  
Available from Sigma-Aldrich MAK224-1KT - Inquire

Available from Sigma-Aldrich.®ion=US