Wroński M
Department of Chemical Technology and Environmental Protection, University of Lódź, Poland.
J Chromatogr B Biomed Appl. 1996 Feb 9;676(1):29-34. doi: 10.1016/0378-4347(95)00404-1.
The essential steps in the assay included electrolytic reduction of disulphides, neutralization, extraction of thiols with 0.1 M tributyltin hydroxide in octane, stripping of the extract with 2% acetic acid, fixing the washed-out amino thiols to a cation exchanger, elution with 2 M hydrochloric acid, oxidation with bromine and evaporation. The remaining octane extract was decomposed by dodecanethiol, the mercapto acids were washed out, oxidized with bromine and evaporated. Both residues were dissolved in water and analysed using capillary isotachophoresis at pH 3.0. Cysteamine was extracted from reduced urine at ca. pH 10, decomposed by dodecanethiol and re-extracted to boric acid followed by determination as a cation. The presence of the following thiols in urine has been confirmed: mercaptoacetic acid, 3-mercaptolactic acid, 2-mercaptopropionic acid, acetylcysteine, mercaptoethanol, cysteine, homocysteine and an un-identified amino thiol. Cysteamine and 3-mercaptopropionic acid could not be detected. Captopril, homocysteine and cysteine were determined quantitatively.
二硫化物的电解还原、中和、用0.1M氢氧化三丁基锡的辛烷溶液萃取硫醇、用2%乙酸反萃取提取物、将洗脱的氨基硫醇固定到阳离子交换剂上、用2M盐酸洗脱、用溴氧化并蒸发。剩余的辛烷提取物用十二烷硫醇分解，巯基酸被洗脱，用溴氧化并蒸发。两种残留物均溶于水，并在pH 3.0条件下使用毛细管等速电泳进行分析。半胱胺从约pH 10的还原尿液中萃取出来，用十二烷硫醇分解，再萃取到硼酸中，然后作为阳离子进行测定。已证实尿液中存在以下硫醇：巯基乙酸、3-巯基乳酸、2-巯基丙酸、乙酰半胱氨酸、巯基乙醇、半胱氨酸、同型半胱氨酸和一种未鉴定的氨基硫醇。未检测到半胱胺和3-巯基丙酸。对卡托普利、同型半胱氨酸和半胱氨酸进行了定量测定。
