A bioluminescence assay for total 3 alpha-hydroxy bile acids in serum using immobilized enzymes.
A bioluminescence assay for bile acids was developed using a co-immobilized 3 alpha-hydroxysteroid dehydrogenase, diaphorase, and bacterial luciferase. The assay was specific for bile acids containing a free 3 alpha-hydroxyl group, as well as androsterone. Light output was linear over a bile acid concentration range of 1-20 000 pmol. Intra-assay precision was 6.2-8.2% and the recovery of added standards was 92-110%. Comparison of results using the bioluminescence assay with those using gas liquid chromatography revealed an excellent correlation (r = 0.99, n = 31). Since the bioluminescence assay is rapid, sensitive, specific, and uses inexpensive reagents, it appears to be an ideal method for the measurement of total bile acids in serum.
D015096:3-Hydroxysteroid Dehydrogenases; D001647:Bile Acids and Salts / Q000097:blood*; D055598:Chemical Phenomena; D002621:Chemistry; D002849:Chromatography, Gas; D008058:Dihydrolipoamide Dehydrogenase; D004800:Enzymes, Immobilized; D006801:Humans; D008156:Luciferases; D008163:Luminescent Measurements
D001647:Bile Acids and Salts; D004800:Enzymes, Immobilized; D015096:3-Hydroxysteroid Dehydrogenases; D008156:Luciferases; D008058:Dihydrolipoamide Dehydrogenase