A HIGH-PRESSURE LIQUID-CHROMATOGRAPHIC ENZYMATIC ASSAY FOR GLYCINE AND TAURINE CONJUGATES OF CHOLIC, CHENODEOXYCHOLIC AND DEOXYCHOLIC-ACID IN SERUM

Google Scholar
  • 1 January 1982
    • journal article
    • research article
    • Vol. 42  (5) , 455-460
Abstract
An assay for glycine and taurine conjugates of choline, chenodeoxycholic and deoxycholic acid in serum by a high pressure liquid chromatographic-enzymatic system is presented. The bile acids were extracted from serum by a reverse-phase liquid chromatographic process with an octadecylsilane column. The bile acid conjugates were separated on a .mu.Bondapak C18 column with methanol/KH2PO4 buffer, 20 mmol/l, pH 5.3 as a mobile phase in < 30 min at a flow rate of 1.4 ml/min. The bile acid fractions were measured by enzymatic fluorometry using a 3.alpha.-hydroxysteroid dehydrogenase-diaphorase system. Recoveries ranged from 82-96%, coefficients of variation were from 5-15% and detection limits were from 0.03-0.08 .mu.mol/l. Mean serum concentrations ranged from 0.10-0.39 .mu.mol/l in the fasting state and from 0.29-1.55 .mu.mol/l postprandially in subjects with normal liver function.