Analysis of conjugated bile acids in human serum by high-performance liquid chromatography using post-column enzyme reaction and off-line fluorimetric determination.
An assay for glycine and taurine conjugates of cholic (GC, TC), chenodeoxycholic (GCDC, TCDC), deoxycholic (GDC, TDC) and ursodeoxycholic (GUDC, TUDC) acids in human serum by high-performance liquid chromatography using an immobilized 3-alpha-hydroxysteroid dehydrogenase post-column reaction and an off-line spectrofluorimetric detection is described. The detection limit is 0.2 microM. Recoveries of extraction and chromatographic analysis range from 89.50 to 91.68% and from 96.75 to 98.83%, respectively. The analysis in serum reveals a coefficient of variation for reproducibility between 16.2% for GC and 4.7% for TDC; inter-assay variability ranges between 4.06% for TCDC and 8.24% for TUDC.
D001647:Bile Acids and Salts / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D004800:Enzymes, Immobilized; D006801:Humans; D006913:Hydroxysteroid Dehydrogenases; D007202:Indicators and Reagents; D008103:Liver Cirrhosis / Q000097:blood; D008297:Male; D013050:Spectrometry, Fluorescence
D001647:Bile Acids and Salts; D004800:Enzymes, Immobilized; D007202:Indicators and Reagents; D006913:Hydroxysteroid Dehydrogenases