I am working on the characterization of the lipids produced by E.coli (ATCC 25922) under different temperature conditions for their eventual use in a leakage assay. Could anyone suggest a protocol to do this using TLC or GC. As far as TLC is concerned can you suggest materials to use for standardization ?
Sarah,
for classes separation of lipids you can use SEC - SDVB column, THF as an eluent.
For detailed fatty acids composition, you should make FAME. Check Supelco or other vendors - they have ready applications.
regards,
G
Hello Sarah,
you may use TLC to separate phospholipids and perform analysis by ESI or MALDI MS.
Have a look to these two articles:
Article Molecular species of phospatidylethanolamine from continuous...
Article Analysis of phospolipids and glycolipids by thin-layer chrom...
Hope it helps.
Article Lipid Compositions in Escherichia coli and Bacillus subtilis...
Article Isolation and Chemical Characterization of Lipid A from Gram...
http://jb.asm.org/content/192/3/643.full
AOCS (American Oil Chemists Society) has a lipid library. I would try FAME by GC-FID (Fatty Acid Methyl Esters) since fatty acids are not volatile unless they are derivatized and they have a 'very' poor UV spectrum.
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