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Determination of Total Glutathione in Yeasts by

High-Performance Liquid Chromatography with Dansylation

Yunfeng Ma, Fu Xiang, Wenwen Jin, and Longjiang Yu*

Institute of Resource Biology & Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074 China.

E-mail: yulongjiang@mail.hust.edu.cn

* Author for correspondence and reprint requests

Z. Naturforsch. 65 c, 391 – 394 (2010); received December 14, 2009/January 27, 2010

A method to determine the content of total glutathione (GSHt) was introduced based on high-performance liquid chromatography (HPLC) with dansylation. The minimum detection concentration of GSHt was 0.5 ȝg/mL and the measurable range 1.0 – 300 ȝg/mL. GSHt in yeasts was obtained by hot-water extraction, GSH complete autoxidation to oxidized glu- tathione (GSSG) in alkaline solution and purifi cation by thin-layer chromatography (TLC).

The quantitative determination of GSSG was derived by dansyl chloride at pH 9.5, 60 ºC for 60 min and assayed by HPLC. GSHt in Saccharomyces cerevisiae is higher than in Candida rugosa and Candida utilis. S. cerevisiae can be chosen as the better target for mutagenesis and industrial scale.

Key words: Total Glutathione, Yeast, HPLC

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