[Determination of L-cysteine in enzymatic reaction mixture with pre-column derivatization by high performance liquid chromatography].
A new sensitive high performance liquid chromatographic method for the determination of L-cysteine in an enzymatic reaction mixture using ultra violet spectrometric detection was developed. The sample reacted with 5,5'-dithio-bis-nitrobenzoic acid (DTNB) and was analyzed on a Shimadzu VP-ODS column at room temperature, using gradient elution with detection at 330 nm. The L-cysteine chromatographic peak was determined in comparison with derivatives of 2-mercapto ethanol and dithiothreitol. The linear range was 5-950 micromol/L. The recoveries were 99.7%-100.5% and the relative standard deviations (RSDs) were less than 1.3%. The detection limit was 0.8 micromol/L. The method is simple and accurate.
Liu
Zhong
Z
College of Life Science, Nankai University, Tianjin 300071, China. liuzhong@eyou.com
Yang
Wenbo
W
Jin
Yongjie
Y
Bai
Gang
G
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003545:Cysteine / Q000032:analysis* / Q000737:chemistry; D004228:Dithionitrobenzoic Acid / Q000032:analysis; D004229:Dithiothreitol / Q000032:analysis; D008623:Mercaptoethanol / Q000032:analysis; D056951:Photoelectron Spectroscopy / Q000379:methods
D008623:Mercaptoethanol; D004228:Dithionitrobenzoic Acid; D003545:Cysteine; D004229:Dithiothreitol