Microanalysis of bile acid in human liver tissue by selected ion monitoring

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Abstract

A method of microquantitative determination of bile acid in 5–30 mg of human liver tissue was developed. Bile acids were converted to their ethyl ester dimethylethylsilyl ether derivatives and analyzed by capillary gas chromatography-selected ion monitoring, using [2H4]-lithocholic (LCA), [2H5]deoxycholic (DCA), [2H4]chenodeoxycholic (CDCA), [2H4]-ursodeoxycholic (UDCA), and [2H3]cholic (CA) acids as internal standards. Precision and reproducibility of the present method were tested using surgically obtained liver specimens. The results were statistically analyzed according to one-way layout and the orthogonal polynomial equation. Bile acids except LCA were determined with 2.3 to 11.4% of the coefficient of variation. Recoveries of conjugated bile acids ranged from 72.2 to 96.0% with a mean of 84.3%. The amount of bile acids present in histologically normal liver specimens (n = 10) was found to be 29.56 ± 8.62 μg/g liver. The relative compositions (%) of CDCA and CA were 38.8 ± 8.9 and 41.1 ± 11.0, respectively.

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