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Under recovery of FAMES

Discussions about GC and other "gas phase" separation techniques.

4 posts Page 1 of 1
I am running the analysis of fish and evening primrose oils by the AOCS method for marine oils (BF3/Methanol derivatisation). In performing some parallel testing with another lab we are getting an under recovery, this is most apparent on the C14 to C18:2 peaks. Are these in anyway mors prone to degradation or harder to derivatise?

Or has anyone got any other ideas.

Thanks

They are more stable than the C18:x, C20:x, C22:x, highly unsaturated FAMES, so your losses are probably due to either volatility or injector discrimination.

You can check for discrimination by using one of the standard AOCS FAME oil test mixes, or prepare you own. If you are getting higher unsaturated and long loner oils, I'd want to ensure that the other lab didn't have analytical problems by using a reference oil or FAME mix, as it's more usual that the unsaturates are lost due to column activity etc.

EPO should be easy, I did 100s of samples in the 1990s, and I derivatised using the quick procedure of Bannon ( it's an ambient temperature methanolic KOH method which takes about 6 minutes a sample, and you can do about six at a time in small 4ml screw cap vials - I think we used C19:0 as the Int std ). Bannon later reported errors can be introduced by the method, but I found that really good mixing ( vortex mixer ) gave quick and accurate results for most oils, including several marine oils.

These days, I suspect there are standard methods for EPO, especially as the oil is sold based on Gamma Linolenic Acid content. I'd look for one of those, or use a second method to check your derivatisation.

The AOCS marine oil method should work for most marine oils, remembering the exclusions listed in the scope of the method.

Please keep having fun,

Bruce Hamilton

If your issues are loss related, use of an internal standard may be a more reliable fix than tweaking of your derivitisation & extraction processes (doing them well is always a plus, but depending on how many people have to run this, training all lab rats to an acceptable level may not be practical).
Thanks,
DR
Image

I assume the method used was:-

AOCS Official Method Ce 1b-89 ( revised 2005 )
Fatty Acid Composition by GLC Marine Oils

Which, I think, uses C21:0 as Internal standard.

Please keep having fun,

Bruce Hamilton
4 posts Page 1 of 1

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