Extraction, Purification And Determination Of Bio-active Polyphenols In Cassia Nomame

Plant polyphenols are widely applied to pharmaceutical, food and cosmetic industries, since they contain a variety of biological activities, such as anti-oxidation, anti-free radicals, anti-ultraviolet and so on. Cassia nomame, called shui zao jiao, shan cha yei, shan bian dou etc. in China, has a long history of utilization in folk. It is used worldwide for the treatment of icterus, migraine,constipation etc., due to its abilities of diuretic, antidote, and so on. The modern pharmacological and phytochemical researches proved that, beside the common characters of other plant polyphenols, Cassia nomame polyphenols, which are a novel sort of plant polyphenolic compounds, posse a strong ability of inhibiting the lipase activity, being able to prevent and reduce obesity efficiently. Therefore, Cassia nomame has a large potential development prospect. Up to now, the major researches about Cassia nomame have been focus on the aspect of pharmacology, however, there is none of report on the extraction, isolation, purification, or determination of Cassia nomame polyphenols. The present studies took an investigation into these fields, and established a set of methods, which are economic and safe, and are suitable to industrial production, for extraction, isolation, and quantitative determination of Cassia nomame polyphenols.The present studies developed a method, with sufficient stability, precision and ruggedness, for quantitative determination of the total content of Cassia nomame polyphenols via UV-vis spectrophotometry, which was the most effective quality control mean to the industrial production of Cassia nomame, so far. The result showed the optimum determination wavelength was A =760nm. A good linearity relationship (correlation coefficient: 0.9990) between the degree of absorbance and the concentration of analyte was obtained, within the range of 0.0004~0.0032mg/ml. The average recovery rates were 99.3%, 99.5%, 99.6%, with its RSD 1.01%, 1.02% and 1.01% (n=5) respectively. By measuring the content of total polyphenols of Cassia nomame on different developing stages, we found that the best harvest period for Cassia nomame was on the fruiting stage.In the extraction experiment, adoption the EtOH , a safe solvent without poison or contaminate, as leaching solvent, which meets the requirement for production of health food and pharmaceutical prducts. At fist the conditions for the extraction were studied by single factor assays as guided by the content of total polyphenols presented in the extract. Then the conditions were optimized, through orthogonal experimental design and variance analysis, with the consideration of cost of production. Thus, the optimum conditions for the extraction of Cassia nomame polyphenol were determined, which were alcohol concentration 70%, temperature 70 癈, extraction time 5 hours and the ratio of solid to liquid as 1:10.The approach of fraction and purification of herbal extract on macroporous resin has attracted more and more interest, with a broad applicability future. The present studies implemented macroporous resin to enrich Cassia nomame polyphenols, bearing the merits of simple instrument, stable quality, low cost and easy to operation. According to adsorption mechanism of macroporous resin, and based on physic and chemical properties of macroporous resin and adsorbent character, static adsorption test and static de-adsorption test were carried out to screen the best macroporous resin for the Cassia nomame polphenols, then the single factor tests, orthogonal experimental designs and variance analyses were applied to optimize the manipulation parameters of macroporous resin. The results indicated that macroporous resin S-8 possessed the strongest adsorption ability to Cassia nomame polyphenols among the resins studied, in addition of an easy de-adsorption property. The optimum adsorption conditions were A3B2C3, i.e. feeding rate 2.5BV/h, concentration of feed 4mg/ml, pH value 6.5. And the optimum elution conditions were A2B3C2D], namely elute volume 3.5BV, elute concen...