To establish a method for concentration determination of puerarin in rats，and to study pharmacokinetics of puerarin in laboratory by this method，and to provide reference for formulation and selection of clinical puerarin medication scheme.Methods:Methods for determination of puerarin content by high performance liquid chromatography(HPLC)reported in literature were analyzed.Chromatographic conditions were established.Chromatographic column：Acclaim C18 chromatographic column(5 μm，4.6 mm×150 mm); mobile phase：methanol:water; detector：Modle 500 UV detector; detection wavelength：250 nm; chromatographic workstation：N2000 dual channel workstation.Puerarin standard solution，external standard solution，plasma sample solution for detection and blank control solution and other solutions were prepared.Gradient elution was used respectively.Specificity，linear relationship，recovery rate，precision and stability of the methods were validated.Twelve clean rats，half male and half female，were randomly divided into 2 groups.The group treated by intraperitoneal administration and gavage with lower limit of recommended dose of puerarin in the pharmacopoeia was taken as low dose group，and the group treated by intraperitoneal administration and gavage with upper limit of recommended dose of puerarin in the pharmacopoeia was taken as high dose group.Orbital venous blood was collected immediately，0.25 h，0.5 h，0.75 h，1 h，1.5 h，2 h，4 h，6 h and 8 h after the gavage.Plasma puerarin concentration was detected by the validated method of HPLC separation and detection.Concentration-time data of puerarin were processed by DAS 2.0 software.Drug-time curves were drawn and pharmacokinetic index was calculated and compared between the groups.Results:1)Under the proposed chromatographic conditions，retention time of main components in rat plasma blank solution，puerarin control solution and puerarin plasma sample solution was significantly different，and no overlap occurred，indicating that the method was specific.2)Different concentrations of puerarin control products were added into rat plasma，and there was a good linear relationship between the concentration and peak area.The regression equation was Y=3.957×106X-697.329，r=0.998 3; detection limit was 0.055～0.660 μg/mL.3)Recovery rate：the recovery rate of puerarin control products in different concentrations was(103.110±0.521)%，and there was no significant difference in the recovery rate between the high dose group and the low dose group(P>0.05).4)Precision：intraday precision was 3.14%，and day to day precision was 5.26%.5)Stability：within 12 h，relative deviation(RSD)of puerarin content between samples placed at different times was 3.95%.6)The drug-time curves of rats with different doses of gavage conformed to two-compartment model.7)After the gavage of puerarin，the AUC0-t，Cmax，MRT0-t and CL of rats with different doses were significantly different(P<0.05).Conclusion:It is feasible to detect puerarin in rats by high performance liquid chromatography.Good efficacy of drug absorption and metabolism can be achieved by using low dose medication scheme，while high dose can affect puerarin absorption.