Pharmacokinetics In Bandicoot For Cinobufagin And Bufogenin Of Delisheng Injection

objective: To establish a HPLC method to determinate the blood concentration for Cinobufagin and Recibufogenin of Delisheng injection in bandicoot. Through animal test make the research to the dynamic change of the concentration for Cinobufagin and Recibufogenin of Delisheng injection in bandicoot to reserch its pharmacokinetics feature. Methods: Use HPLC method detect the concentration for Cinobufagin and Recibufogenin of Delisheng injection in bandicoot. The analysis was performed on a phenomenex C-18 column(250×4.60mm,5μm). The mobile phase consisted of acetonitrile- warter(55:45,V/V), and the flow rate was 1.0 ml·min-1. The column temperature was 35℃and detection wavelength was 296 nm. Norethisterone was chosen as the internal standard. Results: The linear range of Cinobufagin was 0.005~1.000mg·L-1. The linear relation was fine, r = 0.9996, n=7. The linear range of Recibufogenin was 0.010~3.000mg·L-1. The linear relation was fine, r = 0.9997, n=8. The resolution between Cinobufagin and Recibufogenin was higher than 1.5 phase. The results showed that the resolutions between the adjacent components met the pharmacopoeia requirements with good linearity and reproducibility. The average extraction recovery rates in plasma of Cinobufagin and Recibufogenin were 82.3% and 80.2%. The extraction recoveries of these drugs were all higher than 80%. The average blank application of sample recovery rates in plasma of Cinobufagin and Recibufogenin were 99.5% and 102.6%. Use three concentrations of Delisheng injection injected into bandicoot through caudal vein. Through DAS 2.1.1 pharmacokinetics software analysised, the average half life times(t1/2α) of Cinobufagin and Recibufogenin were 0.0905h and 0.4787h, and the same time blood drug level descented rapidly. That might means both of them could distribute rapidly in tissues'acceptors. The t1/2αof Cinobufagin and Recibufogenin would descent with the increase of concentration, because of the drug acceptors saturated gradually. Central room distribution volumes(V1) of Cinobufagin and Recibufogenin were 1.0725L·kg-1 and 1.9710L·kg-1, and increased with the increase of drug concentration. That might means the number of the the drug acceptor was large, and the drug would accumulation in tissues with the increase of drug concentration. The statistical contrasts between high and middle concentrations of Cinobufagin, between low and high concentrations of Recibufogenin and between low and middle concentrations of Recibufogenin might stand for unsaturation of clearance in liver and kidney. The statistical contrasts between high and middle concentrations of Recibufogenin might means saturation of clearance in liver and kidney. So it was essential to monitor the liver and renal function in clinic use the Delisheng injection. Conclusion: This method is simple, rapid, accurate and sensitive. It can be applied to determine the concentrations for Cinobufagin and Recibufogenin of Delisheng injection in bandicoot plasma simultaneously and rapidly and can be thus suitable for use in monitoring therapeutic drugs, studying pharmacokinetics and measuring bioavailabilities.