Study On Components And Quality Of CMR

The first aim of the present study is to optimize the extraction condition for flavonoids from Chrysanthemum morifolium Ramat, and observe the various trends of different components (flavonoids, chlorogenic acid, free-luteolin, total sugar, amino-nitrogen, chlorogenic acid and essential oil ) in Chrysanthemum morifolium Ramat. during collection time. The second aim is to investigate the quality of extract from Chrysanthemum morfolium Ramat. The third aim is to obtain purified apigenine-7-p-D-glucoside and luteolin-7-p-D-glucoside for standard.Optimum conditions (bath temperature, extraction hours, ethanol concentration and extraction times) were determined by the orthogonal experiment guided by the flavonoids content in the extract. The order of factors to affect the flavonoids extracion was ethanol concentration > bath temperature, extraction times > extraction hours. The optimum extraction conditions were 75% ethanol, refluence for 1.5 hr at 100 water bath for 3 times. The flavonoids in Chrysanthemum morifolium Ramat. was extracted under the optimum condition and measured by Spectrophotometery. As the collection time defered, the contents of flavonoids descend obviously.Chlorogenic acid was extracted 2 hours by methanol under refluence in water bath and determinated by HPLC in gradient elution of sodium hydrogen phosphate (pH2.70, 0.01M) and methanol. No significant changes were found in the content of chlorogenic acid.In order to measure luteolin content, ODS column was used, the mobile phase was methanol-tetratetrole-isopropyl alcohol (65:7.8:11.7:15.5) at a flow rate of 0.6 ml-min-1, the UV detection wavelength was 360nm. No significant difference of luteolin content was observed among the different Chrysanthemum morifolium Ramat. in 2001 , except that the sample collected first time of 2001 showed the lowest content of luteolin.The content of total sugar in Chrysanthemum morifolium Ramat. extracted by enathol were mesured by Spectrophotometery. Amino-nitrogen was extracted by water at room temperature and tested by formaldehyde's potentiometric titration. Essential oid extracted by steam distillation was weighted. In the collection prophase of 2001, total sugar did not change, but it obviously ascended later.The total sugar did not change markedly in early collection but ascended obviously in later, in contrast, essential oil content did not vary, but reduced in later collection during 2001. However, no obviously variety was showed in content of essential oil and total sugar in 2002. No significant changes were found in the content of amino-nitrogen.On research of the quality of extract from Chrysanthemum morfolium Ramat., identification and determination were proceeded. The ratio of area of flavonoid glucosides, properties and TLC were identified respectively. The ratio of area between apigenin-7-D-glucoside and luteolin-7-p-D-glucoside were analyzed by HPLC, the value were 1.42 to 1.83. The physiochemical properties for extract from Chrysanthemum morifolium Ramat. were identified in the three system ( HCl-Mg, Molish, AlCl3). Flavonoids and sugar were identified according to physiochemical properties of extract. The TLC for extract were also identified in the two solvent system (ethyl acetate-methanol=2:l,chloroform-methanol=4:l).Luteolin-7-p-D-glucoside and apigenin-7-D-glucoside of extract could be identified by TLC. Quantitative analysis, including the content of total flavonoids and index component were tested. Total flavonoids content were analyzed by sodium nitrite-aluminium nitrate-sodium hydroxide and thick sulphuric acid-phenolcolorimetric methods. The flavonoids content of extract were 63.9% to 70.0%. The content of index component in extract was measured by HPLC. ODS column was used, the mobile phase was sodium hydrogen phosphate-90% acetonitrile at a flow rate of 1ml-min-1, the UV detection wavelength was 338nm. The content luteolin-7-p-D-glucoside of different extract were 4.89 % to 6.84%.Two components, luteolin-7-p-D-glucoside and apigenin-7-...