Summary
High performance liquid-chromatographic analyses of individual bile acids (cholic acid, chenodeoxycholic acid, deoxycholic acid, and lithocholic acid), free and conjugated with glycine and taurine, are described. The analyses of free and glycine-conjugated bile acids are based on esterification of carboxyl group of bile acids with O-(p-nitrobenzyl)-N, N′-diisopropylisourea (PNBDI). Moreover, ursodeoxycholic acid, hyocholic acid, hyodeoxycholic acid and 3ß-hydroxy-5-cholenoic acid also are able to analyse by this method.
These bile acids in biological smaple were extracted by an Amberlite XAD-2 column, and separated by DEAE-Sepharose CL-6B into free, glycine- and taurine-conjugated bile acids. After the separation, free and glycine-conjugated bile acids were esterified with PNBDI directly. Because taurine-conjugated bile acids are unable to be esterified with PNBDI, these bile acids were hydrolyzed by NaOH in order to make free bile acids, and then they were esterified. Because the p-nitrobenzyl ester of bile acids has characteristic ultraviolet absorption, these compounds were separated to individual bile acids by high performance liquidchromatography, and detected by an UV-detector. An analysis of individual bile acids in human bile was demonstrated.
Similar content being viewed by others
References
Sandberg DH, et al: Measurement of human serum bile acids by gas-liquid chromatography. J Lipid Res 6: 182, 1965
Roovers J, et al: An improved method for measuring human blood bile acids. Clin Chim Acta 19: 449, 1968
Okuyama S, et al: High performance liquid- chromatographic separation of individual bile acids: free, glycine and taurine conjugated bile acids. Chem Lett 1976, 679
Makino I, et al: A versatile method for analysis of bile acids in plasma. Anal Lett 5: 341, 1972
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Okuyama, S. High performance liquid-chromatographic analysis of individual bile acids: free, glycine- and taurine-conjugated bile acids. Gastroenterol Jpn 14, 129–134 (1979). https://doi.org/10.1007/BF02773584
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02773584