Bile acids. LXXII. High-performance liquid chromatographic analysis of bile acid coenzyme A derivatives.

Abstract The mobilities of coenzyme A and coenzyme A derivatives of cholate, chenodeoxycholate, deoxycholate, lithocholate, and their 5α analogs were studied in reversed-phase high-performance liquid chromatography. With a C 18 Radial-PAK A cartridge (10-μm particles) and a solvent misture of 2-propanol/10 m m phosphate buffer (pH 7.0, 140360), separation of the chenodeoxycholyl and deoxycholyl coenzyme A derivatives was not observed. An increase in ionic strength of the buffer to 50 m m afforded separation, which was markedly augmented with a C 18 Radial-PAK A cartridge with 5-μm particles. Lowering the pH of the buffer to 5.5 did not materially change the separations regardless of the ionic strength. Quantitation was carried out to a lower level of 8.5 × 10 −12 mol.