A liquid chromatography-tandem mass spectrometric(HPLC-MS/MS)method for the determination of pantothenic acid in urine was developed and validated.After centrifugation and dilution,the sample was separated on an ACPUITY UPLC HSS T3(2.1 mm×100 mm,1.8 μm) column produced by the Waters corporation,and ionized in electro spray positive ion mode,then detected using multi reaction monitoring mode.The calibration curve has a good linear range with a correlation coefficient(r) of 0.999 3.The limit of detection is 0.46 ng/mL,the recoveries are in the range of 87.9%-95.3%,and the RSDs are in the range of 2.5%-13.0%.The method has the advantages of high sensitivity and short analysis time,and could be used to analyze pantothenic acid in human urine.