Simultaneous determination of bile acids in rat liver tissue by high-performance liquid chromatography |
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| Affiliation: | 1. Product Planning and Development Department, Consumer Products Marketing Division, Tokyo Tanabe Co. Ltd., 2-6, Nihonbashi-honcho 2-Chome, Chuo-ku, Tokyo 103-8405, Japan;2. Kazusa Research Laboratories, Tokyo Tanabe Co. Ltd., 100-5 Yana Kisarazu-shi, Chiba 292-0812, Japan;1. Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, China;2. Department of Oncology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China;3. State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nanjing University, Nanjing 210093, China;1. Institute of Animal Nutrition and Functional Plant Compounds, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine Vienna, 1210 Vienna, Austria;2. Faculty of Veterinary Medicine, Brawijaya University, Jl. Mayjen Haryono No. 169, Malang 65145 East Java, Indonesia;1. Department of Civil and Environmental Engineering, University of Houston, Houston, TX, 77204, USA;2. School of Civil and Resource Engineering, University of Science and Technology Beijing, Beijing 100083, China |
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| Abstract: | A method for the simultaneous determination of bile acids in rat liver tissue by high-performance liquid chromatography was developed. Without prior fractionation and alkaline hydrolysis, 30 unconjugated, glycine- and taurine-conjugated bile acids were detected by post-column enzymatic reaction and fluorescence detection. They were separated on a reversed-phase column using a linear gradient solvent system of 10 mM tribasic ammonium phosphate–acetonitrile–methanol (44:12:5, v/v/v) and 20 mM dibasic ammonium phosphate–acetonitrile–methanol (2:1:2, v/v/v). The limits of detection were 1–5 pmol, and calibration curves were linear for concentrations ranging between 10 and 4000 pmol per 10 μl injection. This rapid and reliable method is effective for measuring bile acid levels in liver tissue not only of rats but also of patients with hepatobiliary and other diseases. |
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