Journal of Lipid Research, Vol 20, 986-993, Copyright © 1979 by Lipid Research, Inc.
Radioimmunological determination of serum 3 beta-hydroxy-5-cholenoic acid in normal subjects and patients with liver disease
EI Minder, G Karlaganis and G Paumgartner
A radioimmunoassay for the determination of 3 beta-hydroxy-5-cholenoic acid
in human serum has been developed, using 3 beta-hydroxy-5-
cholenoyl-thyroglobulin as immunogen and 3 beta-hydroxy-5-
cholenoylglycyl-125I histamine as radioactive ligand. The association
constant was 6.3 X 10(8) l/mol. Cross reactivity with other bile acids of
human serum was not detectable, but was 5.6% with cholesterol. Serum sample
preparation included extraction of 3 beta-hydroxy-5-cholenoic acid from
serum, solvolysis of sulfates, hydrolysis of conjugates, and separation
from cholesterol by thin-layer chromatography. Serum concentrations of 3
beta-hydroxy-5-cholenoic acid were 0.23 +/- SD 0.12 mumol/l and 0.21 +/- SD
0.09 mumol/l in healthy males and females, respectively. In patients with
primary biliary cirrhosis the serum concentration of 3
beta-hydroxy-5-cholenoic acid and the quotient 3 beta-hydroxy-5-cholenoic
acid over total 3 alpha-hydroxy-bile acids (measured enzymatically) were
significantly higher (P less than 0.02) than in patients with chronic
active hepatitis or alcoholic cirrhosis. Analysis of 17 sera with elevated
concentration of 3 beta-hydroxy-5- cholenoic acid by radioimmunoassay and
capillary gas-liquid chromatography showed a close correlation (r = 0.91,
slope = 0.97) between the results of the two methods.
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