What is an accurate method for fatty acids estimation in an organic phase sample through GC-MS analysis?
According to me,
Derivatization with BF3 or sodium methoxide and then partition with pet spirit or hexane is best method for fatty acid analysis.
Dunstan et al., 2007 should help you. I use the same for algal lipids and works fine for me.
Dissolved lipids in one volume of toluene and two volumes of 1% sulphuric acid in methanol. Use tridecaenoic acid (C13:0) as an internal standard for calculating the amounts of fatty acid. Incubate the mixture at 80 ºC followed by overnight incubation at 50 ºC. Put 5% NaCl solution to the methanolic mixture and extract fatty acid methyl esters (FAME) in hexane. inject 1 microlitre into GC and detect using FID.
For identification of fatty acids, best way is derivatization following methyl esterification (FAME) and then GC-MS analysis. If you are expecting any unusual fatty acid you can also try DMOX derivatization to confirm the position of double bonds.
For estimation, it's better to use GC-FID using uncommon fatty acid as internal standard. You can use C19:0 or C23:0 as internal standard if these fatty acids are not identified in the sample.
the standards used in fatty acid analysis is FA OR FAME and why
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