HPLC Methyl-isocitric acid

WS via methods%40net.bio.net (by novalidaddress from nurfuerspam.de)
Wed Feb 10 16:58:31 EST 2010


Hi Alex,

although the scope of this NG in fact is much wider than just
molecuIar biology, I doubt that it is the best group for getting a
good answer. Dear fellows, I'd be happy to be proven wrong.

A quick web search like http://www.google.de/search?q=methyl+isocitrate+hplc
seems to yield several promising results on the first results page.

To start from scratch, I'd like to throw my 2c worth in:

The actual problem should not be the separation itself, but the
detection of methyl isocitrate, unless you have some sort of mass spec
detector available. You'll probably need to do a pre-column
derivatization: First, extract the negatively charged compounds from
your analyte, e.g by ion exchange, then derivatize the carboxylic
acids into something fluorescent, eg. by coupling to a fluorescent
molecule with a free primary amino function (a bit like the detection
of amino acids and catecholamines). Use pure (probably commercial)
substances as reference.

For a more direct method, you might be better off with GC after making
the substance more volatile, e.g. by methylation. Detection could be
with FID.

Good luck!

Wo



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