Abstract Ninhydrin reacts with some aminothiols to form spiranes adducts whose optical and chromatographic properties have proved to be useful for chiral recognition. Liquid chromatography-mass spectrometry data along with spectroscopic analysis reveal that under certain conditions, in addition to the known single-spirane configuration, the spirothiazolidinic complexes can exist also as double- and mixed double-spiranes. The reaction was exploited to check the enantiomeric purity of two commercially available dosage form of D-penicillamine and to measure the aminothiol concentration in the urine sample from a subject under treatment with the drug. Separation of diastereoisomers was achieved on a C18 column in isocratic mode by using a mixture of an aqueous solution of formic acid (30 mmol/L)/acetonitrile (90:10, v/v) as a mobile phase. Diastereoisomers were detected by a fluorescence detector and mass spectrometer in short times and with a good resolution. Intra- and inter-assay reproducibility were under 4% with an average recovery of 98%. At a LOD of 0.01%, no evidence of the toxic distomer ( l -enantiomer) was found in the biological sample and drugs.

中文翻译：

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茚三酮与氨基硫醇的螺环化反应的液相色谱-质谱研究

摘要茚三酮与一些氨基硫醇反应形成螺旋烷加合物，其光学和色谱特性已被证明可用于手性识别。液相色谱-质谱数据以及光谱分析表明，在某些条件下，除了已知的单螺环构型外，螺噻唑啉络合物还可以双螺环和混合双螺环存在。利用该反应来检查两种可商购的 D-青霉胺剂型的对映体纯度，并测量来自接受药物治疗的受试者的尿样中的氨基硫醇浓度。通过使用甲酸 (30 mmol/L)/乙腈 (90:10, v/v) 的水溶液混合物作为流动相，在 C18 色谱柱上以等度模式实现非对映异构体的分离。非对映异构体可通过荧光检测器和质谱仪在短时间内以良好的分辨率进行检测。批内和批间重现性低于 4%，平均回收率为 98%。在 0.01% 的 LOD 下，在生物样品和药物中没有发现有毒异构体（l-对映异构体）的证据。