Serum bile acid analysis: a rapid, direct enzymatic method using dual-beam spectrophotofluorimetry

Abstract

The direct quantitative measurement of total bile acids in serum has been achieved using an enzymatic fluorescent method with a dual-beam spectrophotofluorimeter. By use of a 3alpha-hydroxysteroid dehydrogenase, oxidation of bile acids with NAD is completed in 200 seconds with the observed NADH fluorescence being proportional to the concentration of serum bile acids. This method is rapid (8 minutes per individual sample), has an intrinsic sensitivity of +/- micronM of total bile acids, requires no sample preparation and less than 0.8 ml of serum. Paired data analysis using enzymatic fluorescence and gas-liquid chromatographic methods gives a correlation coefficient (r) of 0.99 for 34 samples ranging from 2 to 530 micronM.