A high-pressure liquid chromatographic-enzymatic assay for glycine and taurine conjugates of cholic, chenodeoxycholic and deoxycholic acid in serum.
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.
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*
D001647:Bile Acids and Salts; D002793:Cholic Acids; D003840:Deoxycholic Acid; D002635:Chenodeoxycholic Acid; D013654:Taurine; D005998:Glycine