| Summary: | 碩士 === 靜宜大學 === 應用化學研究所 === 93 === Bile acids are the important end products of cholesterol metabolism in the liver. Abnormalities in cholesterol biosynthesis or metabolism are often reflected in the concentrations and the proportion of different bile acids in various tissues, ex. primary biliary cirrhosis, gallstone, breast cancer and large bowel cancer. The determination of bile acids in these tissues is important in the diagnosis of hepatobiliary diseases. Bile acids also have been applied to therapeutic treatments. For example, chenodexycholic acid and ursodeoxycholic acid have been used as an alternative treatment to surgery in patients with gallstone. In this work, a method has been developed for quantitative determination of six bile acids (cholic acid, deoxycholic acid, lithocholic acid, ursodeoxycholic acid, chenodexycholic acid, hyodeoxycholic acid) in biological tissues including pig liver, pig kidney and bovine liver by gas chromatography- chemical ionization tandem mass spectrometry.
In this work, bile acids were determined as the methyl ester-trimethylsilyl ether derivatives with camphor-10-sulfonic acid as the esterification catalyst. The effects of catalysts, reaction times on derivatization efficiency were evaluated and optimized. Interferences from sterols in biological samples were eliminated by CI-MS/MS without additional pretreatment. Analytical characteristics such as the dynamic linear range of the calibration curves, reproducibility, and recovery were evaluated under the optimized experimental condition. These parameters and the limit of detection 0.4-1.6ng/ml (S/N = 3) indicated that the proposed method was feasible for the determination of trace bile acids in biological samples.
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