Chromatography Forum

Hello everyone
I am trying to methylate/esterify the fatty acids present in muscle phospholipids.
1st I digest the phospholipid to release the fatty acid in C-2 position and then esterify it.
I follow the method from "Direct esterification" published by Lepage and Roy, which includes addition of methanol:benzene 4:1 and incubation for 1 hr at 100C. Then neutralized with K2CO3 and the top benzene layer is used for GC analysis.


The problem am facing is..........this method esterifies both the fatty acids present in C-1 and C-2 position of Phospholipids whereas my objective is to esterify only the enzyme digested fatty acid.

Can someone suggest me a method.
Thanks

Sankella

Diazomethane should do the trick.
By the way where did you see this use of benzene, many labs have banned benzene from their labs in such unnecessary applications.

Here is a web site with lots of good information on derivatization and GC analysis of fatty acids.

http://lipidlibrary.aocs.org/GC_lipid/gc_lip.html

Thanks guys!