Abstract
Objective — To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the rapid determination of tauroursodeoxycholic acid (TUDCA) in bear bile powder. Method — The chromatographic conditions were optimized. A Hypersil GOLD chromatographic column (Thermo Fisher, 100 mm×2.1 mm) was used with the mobile phase of methanol:water (0.1mmol/l ammonium acetate + 0.1% formic acid) = 75:25 for isocratic elution; the column temperature is 30.0℃; the flow rate is 0.2mL/min; the injection volume is 5uL; the negative ion scanning mode and the select reaction monitoring are used. Through the compound optimization function of the instrument, the two characteristic ion pairs of TUDCA are m/z: 498.5/124.0 and m/z: 498.5/79.8 (quantitative ion pair). Results — The detection limit of this method was 5ng/mL, and the linear range was 20~1000ng/mL (r=0.9993). The same sample was measured in parallel with the high performance liquid chromatography (HPLC/UV) method, and the analysis results were basically the same. Conclusion — Compared with HPLC/UV, HPLC-MS/MS is simple, more sensitive and accurate for the determination of TUDCA in bear bile powder. The method can be used in the quality assessment and process control of bear bile powder.
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