| Determination of Pinoresinol Diglucoside within Eucommia ulmoides Oliver, Eucommia ulmoides Oliver leaves, Eucommia ulmoides Oliver skins and Eucommia ulmoides Oliver mushroom; research on fingerprint of Eucommia ulmoides Oliver medicine in different collecting time, and research on extraction and purification of adsorption resin.Use HPLC method for determining the Pinoresinol Diglucoside within Eucommia ulmoides Oliver, Eucommia ulmoides Oliver leaves, Eucommia ulmoides Oliver skins and Eucommia ulmoides Oliver mushroom; content analysis of Pinoresinol Diglucoside from different part of light skin Eucommia ulmoides Oliver of different origins, and of four types of Eucommia ulmoides Oliver variation. The chromatographic conditions adopted was:chromatographic column: Diamonsil-C18(5μ m,150mm x4.6mm); Mobile phase:methanol-water (23:77); Flow rate:1.0ml/min; The column temperature:room temperature; Detected wavelength:277nm; sample size:5ul. Through the investigation of the methodology, this method was suitable for determination of Pinoresinol Diglucoside within Eucommia ulmoides Oliver, Eucommia ulmoides Oliver leaves, Eucommia ulmoides Oliver skins and Eucommia ulmoides Oliver mushroom.Pinoresinol Diglucoside contents of Eucommia ulmoides Oliver, Eucommia ulmoides Oliver leaves, Eucommia ulmoides Oliver skins and different variants are:0.241%,0.0434%,0.36%,0.20%,0.26%,0.23%; No Pinoresinol Diglucoside was found in Eucommia ulmoides Oliver raw skin and mushroom; obvious differences was found between differnet variants. Light skin type raw skin has the lowest content of6.3%, and cracked skin type raw skin the highest of13.6%.Through fingerprint research on Eucommia ulmoides Oliver medicine at different harvest time, review the effect on Pinoresinol Diglucoside yield using different extraction methods; Through the investigation of the material to liquid ratio, three main factors influencing the extraction effect were found:ultrasonic time, ethanol concentration, ultrasonic frequency. According to the orthogonal test, the Pinoresinol Diglucoside yield was calculated.The best technique reached after studying fingerprints of Eucommia ulmoides Oliver at different harvest time is:extraction solvent of60%ethanol is15times of the sample quality, extracting time for40minutes, ultrasonic extraction times for once. Through the fingerprint similarity evaluation, we can see that the similarity between each batch and that between batches and contrast atlas is higher than93%, therefore, the quality of the examined11batches of Eucommia ulmoides Oliver medicinal materials of the same origin at different periods is stable; This experiment has better accuracy, stability, repeatability, so this method can be used to control the quality of Eucommia ulmoides Oliver.In the extraction of Eucommia ulmoides Oliver, compare was made between reflux extraction method mentioned in literature and pharmacopoeia determine method, so effect on Pinoresinol Diglucoside yield was examined. The optimum technological conditions for reflux extraction of Eucommia ulmoides Oliver Pinoresinol Diglucoside is:extraction solvent is60%ethanol, volume ratio between sample quality and extraction solvent isl:15, extracting twice,1.5h each time, extraction temperature80℃. Under these conditions, the yield ratio of Eucommia ulmoides Oliver Pinoresinol Diglucoside is0.3421%. The yield ratio under the pharmacopoeia method conditions is0.3448%.In the research on purification technology of Eucommia ulmoides Oliver, suitable adsorption resin was chosen from D101, AB-8, D4020, NKA-II, X-5through static adsorption and desorption experiment. Through the adsorption and desorption adsorption experiment to choose the best solution, again through the dynamic adsorption chose the best sample conditions; Examine the volume of water used in edulcoration, and edulcorate with selected solution to examine the volume of edulcoration. Finally eluent was yield, and Eucommia ulmoides Oliver extractive was gathered after freeze-drying.The result of Adsorption Resin purification of Eucommia ulmoides Oliver is: D101type of Adsorption Resin was used, the determined purification conditions are:take Pinoresinol Diglucoside extract (0.2g/ml crude), the quality ratio between sample liquid and resin is5:1; With40%ethanol elution agents, the ratio between the volume of water impurity and resin is7:1; the quality ratio between eluent volume and resin than is8:1, eluent to concentrated to small volume, frozen-drying, and yellow Eucommia ulmoides Oliver extract was obtained. D101type of Adsorption Resin is suitable for the purification of Eucommia ulmoides Oliver Pinoresinol Diglucoside; After purification the content of Pinoresinol Diglucoside in the extract is higher than50%. |
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