The Preparation Of The Stachyose Standard Substances With The Guifudihuang Polysaccharide Qc Method

Reference Material means a kind of substance or material with sufficient uniformity and definite characteristic which would be used to standardize metering equipment, appraise the method of measurement and quantitate. With the development of socialisation and globalization of industry, problems such as accuracy in analysis had been solved by reference material, which had ensured the quality of product and increased the manufacturing efficiency. Reference material had been admitted in industry with its development of a century. It had been generally used and developed in many countries.Reference Material of Natural Product Chemistry means a kind of substance or material with activity or toxicity abtained from Troditional Chinese herbs. These reference materials had been usually used to analyze the component in drugs, unify the parameter of work station, calibrate the instrument and evaluate the methods for measuring.The aim of this study was to get a reference material of stachyose, an active components in the root of Rehmannia glutinosa Libosch, by means of separation and analysis used in Natural Product Chemistry. At the same time, method of preparation for the polar reference materials had been researched.Column Chromatography of macroporous adsorptive resin, active charcoal, silica gel and ODS had been used in the research, obtaining 5.07 g of stachyose with purity of 99.8 %. Experiments such as spectrum, precision, homogenicity and stability of the obtained stachyose had been executed according to < Standard for Preparation of Reference Material >. Purity of the stachyose had been evaluated by purity measurement in eight authentic laboratories. Polysacchrides, which exsit extensively in organic body in nature, are composed of monosaccharides by glucosidic bond. Some polysaccharides serve as many physiological functions such as energy storing, structural supporting, defence, antigenic determinant and so on. Polysacchrides had been used in medical science, health food and chemical industry. Researches on polysaccharides initiated at 1950's, which had paid close attention to their pharmacological activity and chemistry. The activity of polysaccharides correlated with their structures closely,including sugar components and their sequence, linkage type, side chains and so on. Researches on quality control of polysaccharides were insufficient, which were accomplished by observing the sugar content, molecular wieghts distribution and composition. These methods could not reflect the information of polysaccharides structure more completely so that they could not be used more effectively for quality control of polysaccharides.In this study, the used polysaccharides were extracted and simply purified from Guifu Dihuang Wan. The polysaccharides were hydrolyzed partially under an appropriate condition of acid hydrolysis, and fingerprint profiles of the acid hydrolyzate could be obtained by HPLC. The aim of the research is to find a new method for quality control of polysaccharides by establishing the fingerprint profiles of polysaccharides. Guifu Dihuang crude drugs of 40.5kg had been extracted, and sufficient polysaccharides were abtained to ensure the uniform and stationariness in the subsequent experiments.In the investigation of hydrolysis procedures for polysaccharides, TFA and sulfuric acid were tested. With crosscut experiments, appropriate condition of acid hydrolysis was found. Polysaccharides were hydrolyzed partially under this condition, so that more fingerprint peaks could be detected by HPLC.The acid hydrolysis solution had been treated with two means to remove the TFA, being blowed or being evaporated to dryness under decompression .The former was better, displaying a better reproducibility.Finally, fingerprint profiles of polysaccharides hydrolyzate from Guifu Dihuang Wan produced by different manufactories were obtained. Microsoft Excel 2002 was applied to calculate the similarity of fingerprint profiles. Information in these fingerprint profiles were abserved by Multivariate Statistical Analysis Software. The results showed that similarity of eight medicine is greater than 0.99 and the similarity of others are 0.92 and 0.94 respectively .