Yamaguchi Y, Seki T, Watanabe S, Hayashi C, Miyai K
J Chromatogr. 1983 Mar 11;273(1):87-94. doi: 10.1016/s0378-4347(00)80925-1.
Urinary sulfated primary bile acids, 7 alpha-hydroxy bile acids, are detected by an enzymatic method using 7 alpha-hydroxysteroid dehydrogenase (EC 1.1.1.-, 7 alpha-HSD) after chromatographic fractionation on Sephadex G-25. Urinary sulfated or glucuronated bile acids are hydrolyzed by beta-glucuronidase/sulfatase (EC 3.2.1.31/EC 3.1.6.1) from Helix pomatia and then released 7 alpha-hydroxy bile acids are detected with 7 alpha-HSD in the presence of beta-ND+, diaphorase (EC 1.6.99.2, from Clostridium kluyveri) and 2-p-iodophenyl-3-p-nitrophenyl-5-phenyltetrazolium chloride. The absorbance of formazan formed during the enzymic reaction is measured at 500 nm. Excretion values of 7 alpha-hydroxy bile acids in normal subjects and in patients with acute hepatitis were compared. This enzymatic detection method for the excretion pattern of urinary 7 alpha-hydroxy bile acids may be useful for clinical diagnosis.
尿中硫酸化初级胆汁酸，即7α-羟基胆汁酸，通过葡聚糖凝胶G-25色谱分离后，采用7α-羟基类固醇脱氢酶（EC 1.1.1.-，7α-HSD）的酶法进行检测。尿中硫酸化或葡萄糖醛酸化胆汁酸经来自苹果螺的β-葡萄糖醛酸酶/硫酸酯酶（EC 3.2.1.31/EC 3.1.6.1）水解，然后在β-ND +、黄递酶（EC 1.6.99.2，来自克氏梭菌）和2-对碘苯基-3-对硝基苯基-5-苯基氯化四氮唑存在的情况下，用7α-HSD检测释放出的7α-羟基胆汁酸。在酶促反应过程中形成的甲臜的吸光度在500nm处测量。比较了正常受试者和急性肝炎患者中7α-羟基胆汁酸的排泄值。这种用于检测尿中7α-羟基胆汁酸排泄模式的酶法可能对临床诊断有用。
