Separation And Transformation Of Monoterpene Glycosides Of Radix Paeoniae Rubra. And Metabolism Of Cordycepin In Vitro

In the first part,the total paeony glycosides(TPG)and monoterpene glycosides of Radix Paeoniae Rubra.(RPR)were taken as the object of study,and to proceed extract and separate of them,and the biotransformation of paeoniflorin was also studied.Firstly,the extraction process of TPG was optimized.The optimal extraction system was as follows: extraction solvent 70% methanol,extraction time 1 h,ratio of solid to liquid 1:10,after the determination of paeoniflorin content of 424 mg/1 g TPG.The powder of RPR was extracted by the optimum extraction condition.The extract solution was filtered and concentrated.The residue was dissolved into water and extracted by petroleum ether,ethyl acetate,water-saturated n-butanol sequentially.The fraction was concentrated and separated by TLC,silica gel column chromatography and the semi-preparing liquid chromatography.The target compound was further purified and crystallized.Five of them were identified on the basis of spectral analysis and physicochemical evidences: 29.5 g of paeoniflorin,235 mg of oxypaeoniflorin,67 mg of benzoylpaeoniflorin,43 mg of benzoyloxypaeoniflorin and0.70 g of albiflorin.The purity of them detected by HPLC was greater than 95%.The prepared paeoniflorin was subjected to ?-glucosidase transformation,and the de-glucose reaction was carried out to obtain the paeoniflorgenin.In the second part,the liver microsomal metabolism of Cordyceps in vitro was studied.The liver microsomes were prepared from rat liver.The concentration of liver microsomal protein was determined by Lowry method.The concentration was 25.88mg/ml.The systems were incubated at 37 ? under constant temperature,and the incubation system was: liver microsomal enzyme 2.0 mg G-6-P 10.0 mmol/L,G-6-P DH 1.0 IU/mL,MgCl2 4.0 mmol/L,NADP 1.0 mmol/L,NADH 0.5 mmol/L,Cordyceps 1.0 mg/10 mL.The sample of highest conversion rate was selected,and the liquid quality was used to detect.The data was analyzed by related metabolic software and related knowledge.Eight metabolites were identified,in which the molecular formulas of seven metabolites were identified.Among the seven identified formulas,the structural formulas of the four metabolites and one metabolite with glutamine were identified,they were 4-carboxylic acid imidazole glucuronide,3,4,5-trimethoxytetrahydrofuran-2-carboxylic acid glucuronide,C6H5O2 glutamine conjugation,furan-2-ylmethanol glucuronide,oxycordycepin.