title: A high-pressure liquid chromatographic-enzymatic assay for glycine and taurine conjugates of cholic, chenodeoxycholic and deoxycholic acid in serum.
abstract: An assay for glycine and taurine conjugates of cholic, chenodeoxycholic and deoxycholic acid in serum by a high pressure liquid chromatographic-enzymatic system is presented. The bile acids are extracted from serum by a reverse-phase liquid chromatographic process with an octadecylsilane column. The bile acid conjugates are separated on a muBondapak C18 column with methanol/KH2PO4 buffer, 20 mmol/l, pH 5.3 as a mobile phase in less than 30 min at a flow rate of 1.4 ml/min. The bile acid fractions are measured by enzymatic fluorometry using a 3 alpha-hydroxysteroid dehydrogenase-diaphorase system. Recoveries ranged from 82 to 96%, coefficients of variation were from 5 to 15%, and detection limits were from 0.03 to 0.08 mumol/l. Mean serum concentrations ranged from 0.10 to 0.39 mumol/l in the fasting state and from 0.29 to 1.55 mumol/l postprandially in subjects with normal liver function.
mesh_terms: D000328:Adult; D000368:Aged; D001647:Bile Acids and Salts / Q000097:blood*; D002635:Chenodeoxycholic Acid / Q000097:blood; D002793:Cholic Acids / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003840:Deoxycholic Acid / Q000097:blood; D005215:Fasting; D005260:Female; D005470:Fluorometry; D005998:Glycine / Q000097:blood*; D006801:Humans; D008297:Male; D008875:Middle Aged; D013654:Taurine / Q000097:blood*
chemical_list: D001647:Bile Acids and Salts; D002793:Cholic Acids; D003840:Deoxycholic Acid; D002635:Chenodeoxycholic Acid; D013654:Taurine; D005998:Glycine