Murata N, Beppu T, Takikawa H, Otsuka H, Kasama T, Seyama Y
Steroids. 1983 Nov;42(5):575-92. doi: 10.1016/0039-128x(83)90121-6.
A Sep-Pak C18 cartridge was used for purification of bile acids from serum. Three kinds of deuterium labeled internal standards were required for accurate measurement of individual sulfated and nonsulfated bile acids. These internal standards were added to the serum before its application to the cartridge. Separation of sulfated and nonsulfated bile acids was performed on piperidinohydroxypropyl Sephadex LH-20 column chromatography. The nonsulfate fraction was submitted to alkaline hydrolysis, and the sulfate fraction to solvolysis followed by alkaline hydrolysis. Each fraction was converted to the hexafluoroisopropyl-trifluoroacetyl derivatives and quantitated by mass fragmentography. The recovery of each bile acid sulfate was quite satisfactory. In fasting healthy subjects the mean of total nonsulfated bile acids in serum was 1.324 micrograms/ml, and that of total sulfated bile acids was 0.450 micrograms/ml. Sulfated lithocholic acid comprised a large part of sulfated bile acids in healthy subjects.
使用Sep-Pak C18柱从血清中纯化胆汁酸。准确测定单个硫酸化和非硫酸化胆汁酸需要三种氘标记的内标。这些内标在血清应用于柱之前添加到血清中。硫酸化和非硫酸化胆汁酸的分离在哌啶羟基丙基葡聚糖凝胶LH-20柱色谱上进行。非硫酸盐部分进行碱性水解，硫酸盐部分进行溶剂解然后进行碱性水解。每个部分都转化为六氟异丙基-三氟乙酰基衍生物并通过质量碎片分析法进行定量。每种胆汁酸硫酸盐的回收率相当令人满意。在空腹健康受试者中，血清中总非硫酸化胆汁酸的平均值为1.324微克/毫升，总硫酸化胆汁酸的平均值为0.450微克/毫升。硫酸化石胆酸在健康受试者的硫酸化胆汁酸中占很大一部分。
