Gas chromatography of lipids...

[Han]

"DanSolo" <daniel.otoole at ucd.ie> wrote in news:1157103858.444626.195750
@m79g2000cwm.googlegroups.com:

> Hi all,
> We are trying to measure serum levels of lipids, particularly linoleic
> and oleic acid. There's quite a few dietary type papers on this:
> 
> Mattes RD. Related Articles, Links
> Oral exposure to butter, but not fat replacers elevates postprandial
> triacylglycerol concentration in humans.
> J Nutr. 2001 May;131(5):1491-6.
> 
> etc etc. We can't do the GC ourselves, but we'd like to prep everything
> as much as possible before handing the samples over to the experts.
> Unfortunately, I can't get my hands on the lipid extraction reference
> from that paper:
> 
> Glass, R. L. (1971) Alcoholysis, saponification, and the preparation of
> fatty acid methyl esters. Lipids 6: 919-925.
> 
> Could anybody give me that or a similar protocol?
> Thanks!
> 
> (oh, and to all the people posting about ligation nightmares, I feel
> your pain brothers, I feel your pain)
> 
General procedure:
1. Isolate the lipid or lipids you are interested in.
2. Saponify in the presence of an internal standard.
3. Perform GC

Decision making process:
1. Decide which lipids you need to analyze.  Total lipid, neutral lipid, 
triglycerides, cholesterol esters, and/or others. 
2. Talk to the people who are going to do the analyses for you.  They can 
tell you *their* preferred isolation method(s), internal standards and 
saponification reagents and vessels.
3. Ask them also from which companies to buy your reagents etc.

HTH
-- 
Best regards
Han 
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