van Berge-Henegouwen G P, Allan R N, Hofmann A F, Yu P Y
J Lipid Res. 1977 Jan;18(1):118-22.
Methods for hydrolyzing and solvolyzing conjugated bile acid sulfates were compared on reference mixtures of conjugated and unconjugated bile acid sulfates gas-liquid chromatography to assess recovery, and thin-layer chromatography and zonal scanning to define the products occurring after hydrolysis. Conventional methods in which solvolysis preceded vigorous alkaline saponification gave incomplete recoveries. However, essentially complete recovery of primary and secondary bile acid sulfates was obtained with a mild alkaline saponification procedure followed by acidification and extraction into ether, in shich complete solvolysis was shown to occur within 12 hours. Based on these findings, we developed and validated a simple hydrolysis-solvolysis procedure; the method features mild alkaline hydrolysis, acidification to pH 1, and extraction with ether followed by a 1-hour incubation.
在结合型和非结合型胆汁酸硫酸盐的参比混合物上，通过气液色谱法比较了水解和溶剂解结合型胆汁酸硫酸盐的方法，以评估回收率，并通过薄层色谱法和区域扫描法确定水解后产生的产物。传统方法是先进行溶剂解，然后进行剧烈的碱性皂化，回收率不完整。然而，采用温和的碱性皂化程序，随后酸化并萃取到乙醚中，能基本完全回收初级和次级胆汁酸硫酸盐，结果表明在12小时内完全发生了溶剂解。基于这些发现，我们开发并验证了一种简单的水解-溶剂解程序；该方法的特点是温和碱性水解、酸化至pH 1、用乙醚萃取，然后孵育1小时。
