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 developed for the quantitative measurement of 11 3 alpha-OH bile acids in paediatric serum samples. Bile acids are enzymatically reduced to the corresponding 3-keto compound and then derivatized with the fluorophore dansyl hydrazine. Preliminary fractionation of bile acids is not required. The limit of detection is 0.1 mumol/L using a sample size of 200 microL. One hundred and twenty-three serum samples were analysed by the high-performance liquid chromatography (HPLC) method and the results compared 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 suitable for use in a clinical laboratory for the further investigation of those patients with abnormally high total bile acid levels where quantification of bile acid fractions is required.
James Fairfax Institute of Clinical Nutrition, Children's Hospital, Camperdown, Sydney, Australia.
R F
RF
Goldsmith
M
M
Gruca
M T
MT
O'Halloran
J W
JW
Earl
K
K
Gaskin
D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific); D001647:Bile Acids and Salts / Q000097:blood* / Q000378:metabolism; D002021:Buffers; D002851:Chromatography, High Pressure Liquid; D003619:Dansyl Compounds / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D006834:Hydrazines / Q000737:chemistry; D006868:Hydrolysis; D010084:Oxidation-Reduction; D012015:Reference Standards; D015203:Reproducibility of Results
D001647:Bile Acids and Salts; D002021:Buffers; D003619:Dansyl Compounds; D005456:Fluorescent Dyes; D006834:Hydrazines; C027913:dansyl hydrazine; D015096:3-Hydroxysteroid Dehydrogenases; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)