Study On The Isolation And Purification Of Flavoniods In Lotus Leaf

Lotus leaf is a residual product or a discarded product from traditional lotus industy. The result has been showed from external and internal, that lotus leaf not only contains physiological active ingredients such as flavonoid and alkaloids, but also contains other affects such as to switch on the activity of hormone sensitivity fat enzyme, to promote glycerine ester hydrolysis in lipocyte, to reduce the content of cholesterol in blood, to restrain atherosclerosis, lose weight and health.Lotus leaf was arranged to a kind of natural plant material used both in food and medicine by the bureau of public health in 1991. At present, chinese reseaches are beening chemical composition, content, extraction, and medication function of flavonoid and alkaloids in lotus leaf, and the alkaloids reseach is more than flavonoid. The study of lotus leaf flavonoid has no break through limited by reseach method and technology conditions. To study flavonoid medication function and application will be a trend in flavonoid sufficient use, based on lotus leaf flavonoid pure products or a monomer extraction used macroporous Resin, HSCCC, structure appraisal and content analysis.Based on establishing the methods of total flavonoid by ultraviolet measure and flavonoid examine by HPLC, using rough extraction of flavonoid from louts leaf, the study compare the effects of static adsorption and desorption in XAD-16, DM-130, HP-20, AB-8, D-101 and so on five macroporous Resin methods, and The best technological parameters of resin's adsorption and desorption were obtained. Afert sifting the solvent conditions of HSCCC and the operated parameter, the lotus leaf flavonoid pure products and a monomer extractions supply a scientific basis on industrialization procreation of lotus leaf flavonoid. The main reseach results of this paper were obtained as follows:1. Set up the test method of flavonoids in lotus leaf:1) Aluminum nitrate spectrophoto metric method measured at awave length of 510 nm, the RSD of precision is 0.21%, the RSD of stability is 5.24%, the RSD of repeatability is 0.36 %, the RSD of recovery is 1.69%, these prove that the method is Reliabilty; 2) Method of HPLC (high-performance liquid chromatography):chromatographic column:Kromasil C18 (150mm×4.6mm,5μm); Mobile Phase:acetonitrile add 0.1 % Phosphoric acid:0.01 min(10:90)-10 min(10:90)-30 min (15:85)-60 min(35:65)-70 min(10:90); Temperature of column:30℃; UV detection:measured at awave length of 360 nm and 254 nm respectively. On this condition, the pik of flavonoids in lotus leaf can be separated efective.2. Studies on separating of flavonoids from lotus leaves by macroporous resins: the material is crude extract which contain 5.46% flavonoid, Static adsorption and desorption of XAD-16, DM-130, HP-20, AB-8, D-101. HP-20 resin, one of the best macroporous adsorption resins, becase of its adsorption capacity and amount of desorption. Adsorbability is 79.86%, desorpability is 65.39%. The reslut of Static adsorption and desorption show that, the sequence of the main factors of the effct to separating of flavonoids from lotus leaves are:eluantstr engthgr adient> elution volume> loading quantities> flow rate.useing the method loading quantities is 40 g, flow rate is 2 BV/h, ethanol concentration is 60%, elution volume is 3 BV ect.to purification of flavonoids extracted from lotus leaves,The totle yield of flavonoids in lotus leaves is 87.78%, purity is 43.83%, the puerty is increased 8.03 time, better than the report of document literature obviously.3. Studies on the structural and purification of louts leaf flavonoid by HSCCC: Undle the conditions that solvent system is Ethyl acetate-Methyl alcohol-H2O2, the rotational speed is 850 r/min, flow rate of immobilityphase is 20 mL/min, flow rate of mobile phase is 1.5 mL/min, detection wavelength is at wavelength of 254 nm, two louts leaf flavonoids were extracted,and the purion is more than 98%. So this method is safety, quick and effective. To conclude with above, the method about examining content of lutos leaf flavonoid by HPLC, the method about rough separating yied rate and purity of louts leaf flavonoid by macroporous adsorption resin, the method about separation and purification of lutos leaf flavonoid by HSCCC, have better. Originality and reference value, compared with the same methods.