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A simple and sensitive high performance liquid chromatographic method coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS) was developed and validated for the analysis of aconitine, mesaconitine and hypaconitine simultaneously in rat plasma.Based on this method, we investigated the effect of the compatibility of Radix Aconiti Laterlis and Radix Glycyrrhizae on the pharmacokinetics of the three alkaloids in Rats.The drugs of Radix Aconiti Latedis decocted singly and Radix Aconiti Laterlis and Radix Glycyrrhizae decocted together were orally administered to two groups of rats, for the same dosage of 1.5 g.kg-1 in Radix Aconiti Laterlis.