Separation of bile acids as their phenacyl esters by high-pressure liquid chromatography |
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Authors: | Frans Stellaard David L. Hachey Peter D. Klein |
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Affiliation: | Division of Biological and Medical Research, Argonne National Laboratory, Argonne, Illinois 60439 USA |
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Abstract: | Bile acids have been separated by high-pressure liquid chromatography. The free acids were derivatized to their phenacyl esters by treatment with triethylamine and α-bromoacetophenone. The stationary phase was a C18, Partisil ODS column. A dual-solvent, stepwise gradient system was used for the mobile phase. The method is applied to a human bile sample and shows excellent resolution of the dihydroxy bile acid phenacyl esters. Detection limits for pure derivatized bile acids are 10–20 pmol (5–10 ng), except for the cholic acid derivative, which has a detection limit of 265 pmol. |
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