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.
Goldsmith
R F
RF
James Fairfax Institute of Clinical Nutrition, Children's Hospital, Camperdown, Sydney, Australia.
Gruca
M
M
O'Halloran
M T
MT
Earl
J W
JW
Gaskin
K
K
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)