Specific determination of cysteine and penicillamine through cyclization to 2-thioxothiazolidine-4-carboxylic acids

被引:49
作者
Amarnath, V [1 ]
Amarnath, K [1 ]
机构
[1] Vanderbilt Univ, Med Ctr, Dept Pathol, Nashville, TN 37232 USA
关键词
plate reader; HPLC-UV detection; 1,1 '-thiocarbonyldiimidazole; cysteine; penicillamine; TTCA;
D O I
10.1016/S0039-9140(01)00631-2
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A very simple and highly specific method for the determination of cysteine and penicillamine is presented. Treatment with 1,1'-thiocarbonyldiimidazole in slightly basic solutions converts cysteine rapidly and quantitatively to a very stable derivative, 2-thioxothiazolidine-4-carboxylic acid, which is not formed by thiols or amines. The cyclic derivative has a characteristic UV spectrum with a maximum at 272 nm and it can be quantified by one of two ways. (1) When only inorganic ions and common additives are present, a spectrophotometer or a plate reader capable of handling multiple samples is sufficient to estimate cysteine in the concentration range of 2-150 muM. Penicillamine is determined similarly by cyclization to 5,5-dimethyl-2-thioxothiazolidine-4-carboxylic acid, The method is also applicable to derivatives of cysteine modified only at the carboxyl group. (2) To determine cysteine in complex mixtures, a liquid chromatograph connected to a UV detector is used. The elution is rapid with well-separated peaks for the thiazolidine derivatives. The detection limit is 2 pmole of cysteine or penicillamine per injection and the detector response is linear up to 1 nmole. The usefulness of the method is demonstrated by determining cysteine and penicillamine in capsules and by measuring cysteine in a dietary supplement. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:745 / 751
页数:7
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