Radioimmunological determination of serum 3 beta-hydroxy-5-cholenoic acid in normal subjects and patients with liver disease.
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.
D000328:Adult; D000368:Aged; D000469:Alkaline Phosphatase / Q000097:blood; D001219:Aspartate Aminotransferases / Q000097:blood; D001647:Bile Acids and Salts / Q000097:blood; D001663:Bilirubin / Q000097:blood; D002770:Cholenes / Q000097:blood*; D002793:Cholic Acids; D002849:Chromatography, Gas; D003429:Cross Reactions; D005260:Female; D006505:Hepatitis / Q000097:blood; D006632:Histamine; D006801:Humans; D008104:Liver Cirrhosis, Alcoholic / Q000097:blood; D008105:Liver Cirrhosis, Biliary / Q000097:blood; D008107:Liver Diseases / Q000097:blood*; D008297:Male; D008875:Middle Aged; D011863:Radioimmunoassay / Q000379:methods; D005719:gamma-Globulins / Q000032:analysis
D001647:Bile Acids and Salts; D002770:Cholenes; D002793:Cholic Acids; D005719:gamma-Globulins; C014596:3 beta-hydroxy-delta 5-cholenic acid; D006632:Histamine; D001219:Aspartate Aminotransferases; D000469:Alkaline Phosphatase; D001663:Bilirubin