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Determination of N-acetylcysteine and thioglycolic acid in human urine

机译：人尿中N-乙酰半胱氨酸和巯基乙酸的含量测定

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摘要

A method for the determination of total N-acetylcysteine and thioglycolic acid in human urine is described. Because these compounds are mainly excreted as disulfides, they are first reduced to the free thiols by treatment with tris(2-carboxyethyl)phosphine hydrochloride and then derivatized with 2-chloro-1-methylquinolinium tetrafluoroborate. Separation and quantitation of the 2-S-quinolinium derivatives of the thiols were achieved by reversed-phase ion-pair liquid chromatography with UV-detection at 355 nm. Because the method enables simultaneous determination of other endogenous urinary thiols, e.g. cysteine and cysteinylglycine, amounts of these compounds in urine were also studied. Detector responses were linear over the range covering most practical situations, with correlation coefficients for all four analytes better than 0.999. Recovery and imprecision (as RSD) were within 99.77-102.17 and 0.01-7.79%, respectively. The lower limit of detection was 0.25 mu mol L-1 urine for thioglycolic acid and N-acetylcysteine, and 0.12 mu mol L-1 urine for cysteine and cysteinylglycine. The method was used for analysis of urine samples from 29 healthy individuals to establish reference values for the thiols, normalized to creatinine. 3-Mercaptolactic acid, 2-mercaptopropionic acid, and mercaptoethanol were not present in the urine analyzed.
机译：描述了一种测定人尿中总N-乙酰半胱氨酸和巯基乙酸的方法。因为这些化合物主要以二硫化物形式排泄，所以首先通过用三（2-羧乙基）膦盐酸盐处理将它们还原为游离硫醇，然后用四氟硼酸2-氯-1-甲基喹啉鎓衍生化。硫醇的2-S-喹啉鎓衍生物的分离和定量是通过反相离子对液相色谱在355 nm处进行UV检测实现的。因为该方法能够同时测定其他内源性尿硫醇，例如半胱氨酸和半胱氨酸甘氨酸，尿液中这些化合物的量也进行了研究。检测器响应在涵盖大多数实际情况的范围内是线性的，所有四种分析物的相关系数均优于0.999。回收率和不精密度（以RSD计）分别在99.77-102.17和0.01-7.79％之内。检测下限为：巯基乙酸和N-乙酰半胱氨酸为0.25μmolL-1尿，半胱氨酸和半胱氨酸为甘氨酸0.12μmolL-1。该方法用于分析29位健康个体的尿液样本，以建立相对于肌酐标准化的硫醇参考值。分析的尿液中不存在3-巯基乳酸，2-巯基丙酸和巯基乙醇。

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