Optimization of determination method and difference analysis of Isatidis folium in Chinese Pharmacopoeia by response surface method
OBJECTIVE To optimize the quality standard and content determination method for Isatidis folium in Chinese Pharmacopoeia.METHODS Extraction solvent was optimized.Taking the volume fraction of phosphoric acid in N,N-dimethylformamide(DMF)solution,the extraction method and the liquid material ratio as the investigation factors,and the con-tents of indigo and indirubin as the response values,the Box-Behnken response surface method was applied to analyzing the inter-action among the factors.The method verification,prediction value verification and method comparison were used to verify the method advantages.RESULTS DMF solution containing phosphoric acid was used as the extraction solvent.The influence order of each factor from high to low on the response value was the volume fraction of phosphoric acid,the shaking time and the liquid material ratio.Finally,the optimal conditions for the preparation of the test solution were determined as follows:0.5 g of Isatidis folium,adding 25 ml of DMF solution with 5％phosphoric acid,shaking for 40min.A method for the simultaneous deter-mination of indigo and indirubin by high performance liquid chromatography(HPLC)was developed.Linear relationship,repeat-ability,recovery and durability were all good.CONCLUSION 5％Phosphoric acid in DMF was used as the extraction solvent for Isatidis folium for the first time.It can effectively improve the extraction efficiency and stability of indigo and indirubin,solve the problems of single index and complicated method of the current standard content determination item,and can be used to control the quality of Isatidis folium.
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