Determination of sulfated and nonsulfated bile acids in serum by mass fragmentography.
A Sep-Pak C18 cartridge was used for purification of bile acids from serum. Three kinds of deuterium labeled internal standards were required for accurate measurement of individual sulfated and nonsulfated bile acids. These internal standards were added to the serum before its application to the cartridge. Separation of sulfated and nonsulfated bile acids was performed on piperidinohydroxypropyl Sephadex LH-20 column chromatography. The nonsulfate fraction was submitted to alkaline hydrolysis, and the sulfate fraction to solvolysis followed by alkaline hydrolysis. Each fraction was converted to the hexafluoroisopropyl-trifluoroacetyl derivatives and quantitated by mass fragmentography. The recovery of each bile acid sulfate was quite satisfactory. In fasting healthy subjects the mean of total nonsulfated bile acids in serum was 1.324 micrograms/ml, and that of total sulfated bile acids was 0.450 micrograms/ml. Sulfated lithocholic acid comprised a large part of sulfated bile acids in healthy subjects.
D001647:Bile Acids and Salts / Q000097:blood*; D002779:Cholestasis / Q000097:blood; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D008103:Liver Cirrhosis / Q000097:blood; D008722:Methods; D012015:Reference Standards
D001647:Bile Acids and Salts