Buck Traditional Chinese Medicine Bidens Total Flavonoids Purification And Quality Standards To Establish

In this paper, the process of extraction and purification on the total flavonoides from spanishneedles herb were studied. A rational technological process was gotten, which establish a foundation for the industrialization. The quantificational and qualitative quality control methods on spanishneedles and its total flavonoides fraction had been estabalished and could be used as quality standard.To get the optimized extraction method, the concentration of ethanol, ethanol quantity, times of extraction and extraction time were investigated by using orthogonal design table and the quantity of the total flavonoids as a target index. The optimized extraction conditions were 12 times as much as the volume 80 % ethanol for three successive extraction and 1.0 hour per time.To study the method of removing the impurity with ZTCⅡclarifying agent, some effecting factors such as the adding order of clarifying agent, the amount of clarifying agent, reacting temperature, reacting time and the concentration of solution of sample etc have been investigated. The optimum conditions for the purifying technology with ZTCⅡclarifying agent are as follows: B compoment is added firstly and then A compoment is added subsequently, the amount of B is 120 mg/100ml, and the amount of A is 60 mg/100ml, the reacting temperature is 40℃, the reacting time is 30 min and the concentration of solution of sample is 0.557 mg/ml.To establish a technical process to purify the total flavonoids from spanishneedles herb, the macroreticular resins were used. The optimal resin was selected from six kinds of macroreticular resins which had higher adsorption capacity and easier desorption capacity. The suitable macroreticular resin was screened out according to the result of statical experiment, in which the adsorption rate and the elution rate of the total flavonoids were used as evaluating primary indexes. The experiment showed that D-101 resin was suitable for adsorbing and stripping the total flavonoids from spanishneedles herb, the best purifying conditions are as follows: pH=6.0 in the sample solution, the sample concentration 4.378 mg/ml, chromatography column diameter-high ratio 1:20, and 70 % alcohol as the stripping liquid. By means of the optimal conditions, the dynamic elution rate on flavonoids was up to 86.7 %. The product purity reached 22.4 %, which was 5.5 times higher than that before purification.Adsorption and desorption capacities of three kinds of polyamides for the total flavonoids were compared. The process condition such as the pH value of the sample solution, the concentration of the sample solution,the maximum adsorption capacity, the eluting method was ascertained. Adsorption and desorption abilities of 60-80 mech polyamide for the total flavonoids were the best of the three polyamides. The optimal pH value was 6, the concentration of the sample solution of the total flavonoids was 2.0 mg/ml, and the maximum adsorption capacity for total flavonoids was 11 mg/g. The total flavonoids were eluted by water and 6 BV 20 % ethanol firstly, then by 4 BV 30 % ethanol, lastly by 2 BV 50 % ethanol and 2 BV 70 % ethanol. The result indicated 60-80 mech polyamide was superior to extraction method of solvent in the separation and collection of the total flavonoids from spanishneedles herb, and it was suitable for separating and purifying the total flavonoids.In the paper, quality standards of spanishneedles herb and its total flavonoids fraction had been established. In qualitative investigation, total flavonoids of spanishneedles herb and its active fraction were indicated by chemical way, and rutin and hyperin were identified by microemulsion thin-layer chromatographic method. In quantitative investigation, the UV method was established to determine the content of total flavonoids in spanishneedles herb and its total flavonoids fraction. Rutin was selected as good representative constituent, which be colored by AlCl3-KAc. Moreover, the HPLC method was established to determine the content of rutin in spanishneedles herb and its total flavonoids fraction. All the methods of determination were accurate and feasible.