Establishment And Verification Of Design Space For Leonurus Japonicas Extraction Process Based On Quality By Design

ObjectiveThe purpose of this study is to optimize the extraction procedure of Hydrochloride Stachydrine and leonurine in Leonurus japonicas, to set the technology objectives, to experiment with the key process parameters of extraction procedure- including the amount of water, number of extraction times and extraction, to establish the model and to design space, then based on the model to predict actual extraction efficiency of Hydrochloride Stachydrine and leonurine for guiding the actual production.Methods1 To use RSM(Response Surface Methodology), according to Quality by Design(Qb D) concept, to optimize extraction process of Leonurus Artemisia in Naomai Li particle.2 To determinate the leonurine by HPLC, the mobile phase used was acetonitrile: water(0.4% 1-Octanesulfonic acid sodium salt Monohydrate)=25:75,the flow rate was 1.0ml/min, the detection wavelength was sat at 276 nm,injection volume was 20?l. A TLC Scanner method was set up to determinate the Stachydrine,the developer was made from ingredients with N-butyl alcohol-acetic acid-ethyl ester(8:3:1), ?s=508nm,?R=670nm?3 The Design-expert software was used to analyze the experimental data for building a model that it could show the relationship between the critical qualities attributes and the key process parameters.Results1 According to RSM design, using Design-expert software for experimental design center, 20 trails were been tested to build a model, an equation was given and work out the optimal scheme, Stachydrine hydrochloride = 195.1-23.89*A+10.49 *B-31.15*C +1.66*A*B+0.84*A*C+2.48*B*C+1.18*A2-9.74*B2+2.81*C2;Leonurine=253.8-36.36*A-3.94*B-31.05*C+2.13*A*B+1.53*A*C+2.65*B*C+1.65*A2-7.12*B2+1.28*C2. Fitting correlation coefficient of two equations R2 is 0.7363 and 0.7656 respectively, indicating a good fit between the actual and predicted values, this model could predict the extraction rate of Hydrochloride Stachydrine and leonurine.2 The results showed a good linear relationship with range of stachydrine from 10.09 to 40.36? g(r=0.9977), leonurine from 5.14 to 82.24 ? g/ml(r=0.9994), the average recoveries of stachydrine was 99.48%( RSD=2.34%,n=9), the average recoveries of leonurine was 98.10%( RSD=2.02%,n=9).Conclusions1 The established methods were simple, convenient and practicable, and can be used to determinate the assay of Hydrochloride Stachydrine and leonurine.2 The result of the statistical analysis of established model that P-value of the model was less than 0.05(significant), and the lack of fit values were greater than 0.1(not significant) indicating that the model is reliable and could show the relationship between the factors and the response value. The result that P value of amount of solvent A, extraction times B, extraction holding time C were all less than 0.05 indicating that A, B, C three factors have a significant influence on Y1, Y2 and influence level are sorted A> B > C. But the result that P values of AB, AC, BC were less than 0.05 shows the effect of the interaction of A, B, C three factors for Y1, Y2 were not significant.