Nishiyama J, Kuninori T
Anal Biochem. 1984 Apr;138(1):95-8. doi: 10.1016/0003-2697(84)90774-7.
A simple and rapid method for the determination of nanomole levels of biological thiols is described. The analysis is based on the combination of reverse-phase high-performance liquid chromatography with a postcolumn reaction with 6,6'-dithiodinicotinic acid. Thiols, including cysteine, cysteamine, thiolhistidine, homocysteine, glutathione, penicillamine, ergothioneine, and thiouracil were separated by eluting with 33 mM KH2PO4 at pH 2.2. Glutathione, cysteine, cysteamine, homocysteine, and penicillamine were quantitatively determined with detection limits of 0.1 nmol, while the quantitative detection of thiolhistidine, ergothioneine, and thiouracil was not successful. The method was applied to the assay of glutathione in human erythrocytes and Escherichia coli.
本文描述了一种测定纳摩尔水平生物硫醇的简单快速方法。该分析基于反相高效液相色谱与6,6'-二硫代二烟酸柱后反应的结合。硫醇，包括半胱氨酸、半胱胺、硫醇组氨酸、同型半胱氨酸、谷胱甘肽、青霉胺、麦角硫因和硫脲嘧啶，通过在pH 2.2的33 mM KH2PO4中洗脱进行分离。谷胱甘肽、半胱氨酸、半胱胺、同型半胱氨酸和青霉胺的检测限为0.1 nmol，可进行定量测定，而硫醇组氨酸、麦角硫因和硫脲嘧啶的定量检测未成功。该方法应用于人体红细胞和大肠杆菌中谷胱甘肽的测定。
