Study On The Extraction Of Steroid Saponins In Rhizoma Paridis And Its Pharmacokinetics In Rats

After years of research，it has been proved that Paridis saponin is the main activeingredient of Paridis herb,which has various pharmacological effects. However, themetabolism of Paridis saponins in-vivo have not been reported yet. This articleexplores the pharmacokinetic characteristics of Paridis saponins and its preliminarilymetabolism in rats. The contents and results are as follows:1. Extraction of Paridis herb.A standard curve method has been set up,taking theParidis saponinⅠandⅡas targets.The impact on the percentage of the extractionprocess of the extraction methods and solvents has been investigated.Then,theorthogonal experiment was carried out to optimize the method,in which three factorsand three levels were selected in orthogonal design. With the standard of Paridissaponin’s contents and collection rate of cream, orthogonal experiment determined thebest extraction program by the two methods of analysis of variance and range analysis.At last,it is finalized that the the saponin extraction method is using20volumes of40%ethanol to extract for60minutes.2. Gather the active ingredients from the extracts of Paridis. This chapter comparedthe effects using n-butanol extraction method and macroporous resin chromafographymethod.70%ethanol was used to elute total saponins chromafographed on macro-porous resin,but the results prove its elution capacity is not ideal.N-butanol extractionmethod is better, but the content of saponins is still low.Thus,the purification methodwas optimited to use a macroporous resin adsorption method.The elution method iseluting with5-fold column volume of purified water,60%,80%ethanol respectively.Collect80%ethanol elution eluent,recovery the solvent and obtain Paridis saponinextracts under reduced pressure.3. Establish the HPLC-MS/MS method for pharmacokinetic study of Paridissaponins.A HPLC-MS/MS method was set up to determine the concentration ofsaponins in the plasma. This method is highly specific and not subject to interferencefrom other endogenous substances under specified conditions.It also has a highersensitivity.The blood concentration of the lowest quantitative limit is18ng/ml. There is a good linear relationship for Saponin saponinⅠin the range of18.3~585ng/ml,saponinⅡin the range of18.6~596.7ng/ml.The method of recovery,precision,extraction recovery, stability are all match the requirements of the analysis ofbiological samples examination.4. Study of pharmacokinetic of Paridis saponins extracts by in-vivo injection inrat’s tail. After injecting Paridis saponin extract solution invo rats, plasma sampleswere processed according to the method. The concentration of Paridis saponin inplasma was measured at different time points. The measured data is fit by softwareDAS2.0for calculating major pharmacokinetic parameters. The results show thatparidis saponin I and II are in line with a two-compartment pharmacokinetic model,their clearance rates are all quickly.5. Study of pharmacokinetic of Paridis saponins extracts by oral administration.Give the rat Paridis saponin extract suspension by oral administration.Plasma sampleswere processed in accordance with the method above.The concentration of Paridissaponin in plasma was measured at different time points. The measured data is fit bysoftware DAS2.0for major pharmacokinetic parameters.The bioavailability wascalculated.The results show that Paridis saponins I and II are in line with the one-compartment pharmacokinetic model.There may exist the enterohepatic circulation,and Paridis saponins I, II have an extremely low bioavailability.