DETERMINATION OF CONJUGATED AND UNCONJUGATED SERUM 3-ALPHA-OH BILE-ACIDS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A Simple reverse-phase liquid chromatographic method has been develope d for the quantitative measurement of 11 3 alpha-OH bile acids in paed iatric serum samples. Bile acids are enzymatically reduced to the corr esponding 3-keto compound and then derivatized with the fluorophore da nsyl hydrazine. Preliminary fractionation of bile acids is not require d. The limit of detection is 0.1 mu mol/L using a sample size of 200 m u L. One hundred and twenty-three serum samples were analysed by the h igh-performance liquid chromatography (HPLC) method and the results co mpared with a commercial kit method (Enzabile) presently used in many laboratories for the measurement of total bile acids. There was a good correlation between the two methods (r = 0.96). The method is suitabl e for use in a clinical laboratory for the further investigation of th ose patients with abnormally high total bile acid levels where quantif ication of bile acid fractions is required.