Goto J, Sano Y, Tsuchiya K, Nambara T
Pharmaceutical Institute, Tohoku University, Sendai, Japan.
J Chromatogr. 1988 Mar 4;425(1):59-66. doi: 10.1016/0378-4347(88)80006-9.
The separation of disulphates of cholate, chenodeoxycholate, deoxycholate and ursodeoxycholate and their glyco and tauro conjugates has been carried out by high-performance liquid chromatography (HPLC) on a reversed-phase column. The chromatographic behaviour of bile acid disulphates was dependent on the position of the sulphate and hydroxy groups and the structure of the side-chain. The method has been applied to the separation and characterization of disulphates in biological fluids without prior deconjugation. The disulphate fraction was obtained from a urine specimen by passing it through a Sep-Pak C18 cartridge, followed by group separation by ion-exchange chromatography on a lipophilic gel, piperidinohydroxypropyl-Sephadex LH-20. Subsequent resolution into individual disulphates was attained by HPLC on Radial-Pak A and Cosmosil 5C18 columns. Taurochenodeoxycholate 3,7-disulphate and taurodeoxycholate 3,12-disulphate in human urine were unequivocally identified on the basis of their behavior in HPLC using mobile phases of different pH.
利用反相柱通过高效液相色谱法（HPLC）对胆酸盐、鹅去氧胆酸盐、脱氧胆酸盐和熊去氧胆酸盐的二硫酸盐及其糖基和牛磺基共轭物进行了分离。胆汁酸二硫酸盐的色谱行为取决于硫酸根和羟基的位置以及侧链结构。该方法已应用于生物流体中二硫酸盐的分离和表征，无需事先进行脱共轭处理。通过将尿液标本通过Sep-Pak C18柱获得二硫酸盐部分，随后在亲脂性凝胶哌啶羟基丙基-葡聚糖凝胶LH-20上通过离子交换色谱进行分组分离。通过在Radial-Pak A和Cosmosil 5C18柱上进行HPLC，将其进一步分离为各个二硫酸盐。根据人尿中牛磺鹅去氧胆酸3,7-二硫酸盐和牛磺脱氧胆酸3,12-二硫酸盐在不同pH值流动相条件下的HPLC行为，明确鉴定了它们。
