Studies on steroids. CCXXXIII. Separation and characterization of bile acid disulphates in human urine by high-performance liquid chromatography.
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.
Goto
J
J
Pharmaceutical Institute, Tohoku University, Sendai, Japan.
Sano
Y
Y
Tsuchiya
K
K
Nambara
T
T
D001647:Bile Acids and Salts / Q000302:isolation & purification / Q000652:urine*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D013056:Spectrophotometry, Ultraviolet; D013256:Steroids / Q000302:isolation & purification / Q000652:urine*; D013431:Sulfates
D001647:Bile Acids and Salts; D013256:Steroids; D013431:Sulfates