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aa derivatization with ethylchlororformate

Posted: Fri Jul 15, 2005 5:25 pm
by Guilherme
Hello
somebody has experience with aminoacid derivatization using ethylchloroformate. I'm trying to derivatize homocysteine in body fluids. but nor in the standards I obtaine peaks.
Some help??
Guilherme

Homocisteine derivatization

Posted: Tue Jul 19, 2005 7:50 pm
by josebenjamin
Dear Friend,

I am not clear whether you are trying to use this derivatization for an Amino Acid and then trying to analyze the product by GC. If that is the case, then you are using the wrong derivatization with the wrong analytical method.

FMOC and FLEC derivatizations are used to introduce a chromophore into all secondary and primary NH2 groups. The products usually can be analyzed in very simple HPlC conditions, such as reversed-phase, NaOAc buffer pH 4.5, UV detection.

If you want to analyze your amino acid by GC then you have to use some TMS derivatization.

Good Luck,

josebenjamin

Posted: Fri Jul 22, 2005 9:23 pm
by Guilherme
Dear José,
thank you for your reply, but I'm trying to use Husek's methodology.

- Husek P, Matucha P, Vrankova A, Simek P.
Simple plasma work-up for a fast chromatographic analysis of homocysteine,
cysteine, methionine and aromatic amino acids.
J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Jun 15;789(2):311-22.

- Husek P.
Chloroformates in gas chromatography as general purpose derivatizing agents.
J Chromatogr B Biomed Sci Appl. 1998 Oct 9;717(1-2):57-91. Review.

- Husek P.
Simultaneous profile analysis of plasma amino and organic acids by capillary
gas chromatography.
J Chromatogr B Biomed Appl. 1995 Jul 21;669(2):352-7.

Thank you again.
Guilherme

Posted: Thu Aug 18, 2005 6:49 pm
by CIL-BAHA
(removed)

Posted: Fri Aug 19, 2005 1:10 am
by Guilherme
Dear CIL-BAHA,
what dou you mean?