Simultaneous determination of four bufadienolides in human liver by high-performance liquid chromatography

A high-performance liquid chromatographic method for the simultaneous quantitation of four bufadienolides-cinobufotalin, bufalin, cinobufagin and resibufogenin-in human liver was investigated. The procedure involved solid phase extraction of human liver with an Oasis HLB cartridge coupled with reversed-phase HPLC and photodiode array detection. Recoveries obtained from spiked liver for the bufadienolides were better than 70%. The linearity was studied up to 1.2 mg/kg and the detection limits of the method were 0.4 ng for cinobufotalin and bufalin and 0.5 ng for cinobufagin and resibufogenin. The developed method was successfully applied to a lethal poisoning case.