Capillary gas chromatographic analysis of protein amino acids as their N(O,S)-isobutoxycarbonyl methyl ester derivatives

Biomedical Chromatography : BMC
S MatsumuraM Makita

Abstract

A simple and rapid method for the determination of the 22 protein amino acids by capillary gas chromatography is described. The amino acids were converted into their N(O,S)-isobutoxycarbonyl (isoBOC) methyl ester derivatives and measured by GC with flame ionization detection using a DB-17 capillary column. Arginine was converted into ornithine by arginase treatment prior to the isoBOC reaction. The N(O,S)-isobutoxycarbonylation of amino acids with isobutyl chloroformate was completed within 15 s by sonication in aqueous alkaline media. All of the derivatives were quantitatively resolved as single symmetrical peaks within 9 min by GC on a single column. The detection limits of amino acids were 0.2-4.0 ng per injection, and the calibration curves were linear in the range 0.2-50 micrograms for each amino acid. This method was successfully applied to protein hydrolysate samples. The analytical results of amino acid compositions of several proteins are presented.

References

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Citations

May 17, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Man-Jeong PaikKyoung-Rae Kim
Jun 7, 1996·Journal of Chromatography. B, Biomedical Applications·S MatsumuraM Makita

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