| Formula composed of several herb medicines is the most widely used form of traditional Chinese Medicine (TCM). The therapeutically efficacy of TCM formula is the result of rational combination of those herbs with appropriate proportion. It has been proven that herb used alone can't produce the effect of whole formula. Therefore our research regarded Xuefuzhuyu Formula as an example to investigate some undefined problems, which are very important for TCM research such as separation technique, qualitative and quantitative analysis methods, even the active component screening technology. At the same time, the application of macroporous adsorption resin in separation and active component enrichment was studied. 1 Active components researchThe crude extracted from formula was gotten at first. Then isolation process has been performed with the use of the Macroporous adsorption resin D101. Qualitative analysis and HPLC fingerprint of each fraction were involved in quality control. Meanwhile some methods have been established to analyze some main ingredients (polysaccharide, total flavones etc.) and substances (naringin, paeoniflorin, ferulic acid etc.). Moreover, an animal model was arranged to determine pharmacological activity of components. The result exhibited that two active components (PF, FP) with anti-hyperlipemia effect have been found out and the synthesis effect of themhave been validated. Through the way of compared chemical constitution of relevant components with original formula and herbs, relationship of chemical constituents and the sources of active components have been obtained. The component one (in PF fraction) chiefly belongs to Huoxue medicine while the component two (in FP fraction) comes from Liqi medicine. By this unique strategy, we got successful results with the discovery of active components and identification of their chemical ingredients. At last, we studied the influence of different combinations ofXuefuzhuyu formula on concentration of paeoniflorin and ferulic acid in aqueous extracts, result showed that the concentration of paeoniflorin was increased and ferulic acid decreased in condition of mixed extraction of every herb rather than single herb extraction.2 Preparation process research ExtractionThe orthogonal experiment design L16 (45) was performed to select the optimalcondition of extraction process for targeted components. We have considered the influence of different experiment factors including extraction solvents, extraction times, spending time and temperature, which may significantly affect the extraction rate of targeted components. As a result, the optimum extraction condition to component one is: 40% alcohol for 3 times with 1.5 hours at 80 water of circumfluence , the component two: 40% alcohol for 4 times while each time lasted for 1.5 hours at 80 circumfluence extraction with water. PurificationWe studied the relationship between adsorption capacity of macroporous adsorption resin D101 and the initialization condition of sample concentration, pH of solvent, temperature, flow rate and etc. As a result, the optimal adsorption condition is, for component one: 0.4mg/ml, pH6, the flow rate 2BV/h, 12 times (BV) of 40% alcohol as eluant solvent; for component two: 0.5mg/ml, pH3, the flow rate IBV/h, 1.5 times (BV) of 70% alcohol as eluant.Furthermore, the orthogonal design L16 (45) was used to select the optimum purification process condition. We have studied the influence of purified solvent andit's volume, eluant, amount of eluant, which could affect the contents and recoveries of active ingredients in components. The result shows the optimum purification process of component one was purified with 6 times(BV) 20% alcohol, the flow rate 2BV/h, added sample 60% of max adsorption of resin, 3 times(BV) 40% alcohol as eluant solvent. Under the optical condition, the content and recovery of component one were 46 3.42%and 87. 6?. 45%(n=3), the component two: purified with 8 times(BV) 20% alcohol, the flow rate IBV/h, added sample 60% of Max adsorption of resin, 3...
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