Optimization Of Extraction Technology Of Effective Components In Anti-liver Fibrosis Of Biejiajianwan Prescription And Preliminary Study On Their Correlation With Drug Efficac

Objective:A method for simultaneously determining of Paeoniflorin,Baicalin and Paeonol in Biejiajian pills was established to investigate the optimal extraction process with the parameters.Based on verification of its efficacy,the correlation analysis of pharmacodynamics index and three index components provides system data for subsequent active fractions research.Method:The orthogonal design was used in the extraction,according to the yield rate and content of Paeoniflorin,Baicalin and Paeonol in Biejiajian pills,to optimize extraction process conditions.Pearson correlation coefficient method was performed using SPSS 20.0 to analyze the correlation of inhibition rates of HSC-T6 and else evaluation indexes.Results:1、Chromatographic separation was achieved on an Eclipse XDB-C18(5 μm,4.6 mm×150 mm).The mobile phase A was 0.1%Phosphoric acid in water.The mobile phase B was acetonitrile.Gradient elute conditions:0-10 min 13%A,87%B;10-20 min 30%A,70%B;30-33 min 13%A,87%B;after 33min,13%A,87%B,at a flow rate of 1.0 mL min-1.The UV detection was performed at a wavelength of 240 nm and 274nm.The injecting sample volume was 10 μL.HPLC analysis was performed at 40℃ to simultaneously determine the content of three components.2、Extraction process:The effect of extraction time and the material-to-liquid ratio on extracting technology of Paeoniflorin has a significant difference.The optimal extracting technology of Paeoniflorin was A1B2C2D3 performed by 70%ethanol,1 h extraction for 3 times,with 8-fold volume of water.The effect of extraction time and the ethanol concentration on extracting technology of Baicalin has a significant difference.The optimal extracting technology of Paeoniflorin was A1B2C2D3 performed by 70%ethanol,1 h extraction for 3 times,with 8-fold volume of water.The effect of the ethanol concentration on extracting technology of Baicalin has a significant difference.The optimal extracting technology of Paeoniflorin was A1B3C2D1 performed by 90%ethanol,1 h extraction for 1 time,with 8-fold volume of water.3、The Pearson correlation coefficient of Paeoniflorin content and inhibition ratio of cell was(r)=0.886(P<0.01),which demonstrated that extraction rate of Paeoniflorin and cell inhibition rate had a positive linear correlation.but Pearson correlation coefficient of paeonol,baicalin,and extraction rate(r)=0.313、0.148、0.361(P>0.05),which suggested that there was not a positive linear correlation.The extracting solution of Biejiajian pills had a significant inhibition effect on proliferation of HSC cells.Conclusion:The method for simultaneously determining process of the content of three components in one mobile phase system is fast and easy operation.The optimal extracting technology of Paeoniflorin Baicalin and Paeonol was stable,feasible and reasonable.The pharmacodynamics research showed that extracting solution of Biejiajian pills had a significant inhibition effect on proliferation of HSC cells.Paeoniflorin content and cell inhibition rate had a positive linear correlation,but paeonol,baicalin,and extraction rate not.