A method was established for the simultaneous determination of 20 kinds of bile acids in human serum employing ultraperformance liquid chromatography-tandem mass spectrometry. Chromatographic conditions and sample preparation were optimized to achieve good separation and maximum sensitivity for these analytes. The linearity, accuracy and repeatability of the development method were validated with a series of experiments. Under the optimum conditions, good linearities were achieved in the quantitative range for each bile acid with the correlation coefficients (r ) >0.9901. The limit of detections (signal-noise ratio 3) of the method were in a range from 0.02 to 0.57 nmol/L. The recoveries were in the range of 88.1-109.9%, RSD < 6.12%. This method was successfully applied for the determination of bile acids in a human serum sample with simple operation, high sensitivity and good accuracy, and provides a reference for the clinical determination of bile acid content.
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