NAD^+と固定化3β-ヒドロキシステロイド脱水素酵素を用いる高速液体クロマトグラフィーによる3β-ヒドロキシ胆汁酸とそのグルクロニドの新測定法

概要

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Most of the bile acids in body fluid are conjugated with glycine and taurine, and some of them are esterified as sulfate and glucuronide. Because of poor volatility of amino acid conjugated bile acids, these bile acids can not be determined directly by gas chromatography. Therefore, it is necessary to separate these bile acids and deconjugate amino acids from them prior to determination of the bile acid in gas chromatography. In order to improve this difficulty, the authors developed a new analytical method for conjugated 3β-hydroxy bile acids and its glucuronides by high-performance liquid chromatography (HPLC) combined with immobilized 3β-hydroxy steroid dehydrogenase (3β-HSD) and NAD^+ as a system for detection without pretreatment of fractionation and deconjugation. However, this method requires a simple ethanol extraction of bile acids from biological samples and enzymatic hydrolysis of glucuronides by β-glucuronidase as pretreatment. It is concluded that this method is useful for the elucidation of pathophysiological significance of 3β-hydroxy bile acids in hepatobiliary disease, because of high accuracy, high sensitivity and high specificity of this method.
日本ビタミン学会の論文
1986-04-25