A method for the quantification of quercetin-3’-O-glucoside in rat plasma by high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) was developed and validated.Along with internal standard(carbamazepine),quercetin-3’-O-glucoside was extracted from plasma samples by simple liquid-liquid extraction with ethyl acetate.The mass spectrometry detection was set in multiple reaction monitoring(MRM) mode via positive electrospray ionization(ESI).The chromatographic run time was 3.5 min per sample.The calibration curves were linear(r^2 = 0.9992) with a lower limit of quantification(LLOQ) of 10.625 ng/mL,and the limit of detection(LOD) was 4.25 ng/mL.The intra-and inter-day precision and accuracy,in terms of relative standard deviation(RSD),were all lower than 10.44%.The recovery rate of the analyte and internal standard were higher than 66.80%.After intravenous administration of 10 mg/kg quercetin-3’-O-glucoside,the t1/2 and AUC were(0.02±0.01) h and(1.22±0.28)×10^4 μg/L·h.The method is accurate,stable and sensitive,which is suitable for the pharmacokinetic study of quercetin-3’-O-glucoside in rats.