Study Of Components In Hebei Balmy Chrysanthemum

Chrysanthemum morifolium Ramat. belongs to the large chrysanthemum family. It has been widely used in traditional Chinese medicine. The medical part of this plant is it's capitulum. It has the effects of dispersing the wind-evil, relieving fever, calming the liver and improving eyesight, detoxifcation and antihypertension. Chrysanthemum morifolium Ramat. originated in China. It has been planted in many provinces of our country. There have been many varieties of chrysanthemum because of different environment, growth, raise and management. Presently, there are eight medical chrysanthemums, such as Hang white chrysanthemum, Bo chrysanthemum, Chu chrysanthemum, Gong chrysanthemum, Ji chrysanthemum, Qi chrysanthemum, Yellow chrysanthemum and Huai chrysanthemum in the market in our country. Hebei balmy chrysanthemum is a new member in chrysanthemum family, and it is the only type of aroma chrysanthemum, containing 1.6% essential oil and 5% flavonoid, which are higher than other types of chrysanthemum. Therefore, Hebei balmy chrysanthemum has exploitation value. Our previous studies showed that the extraction of Hebei balmy chrysanthemum has the effect of antihypertension significantly. In order to look for antihypertensive fractions or components, we studied the chemical components in Hebei balmy chrysanthemum systemly and determined the total flavonoid in the 60% ethanol active part in this paper.Objective: 1 To study the chemical components in Hebei balmy chrysanthemum, column chromatogram, preparation of thin layer chromatogram, polymide column chromatogram and sephadex column chromatogram were used, in order to get pure compounds. To determine the structures of compounds by modern spectrum technology such as UV, 1H-NMR, 13C-NMR, DEPT, MS. Those studies are helpful to investigate antihypertensive fractions or components.2 Determine the flavonoid in the 60% ethanol active part.Methods: 1 Preliminary test of composition of Hebei balmy chrysanthemum: air-dried capitulum 100g were extracted with 95% ethanol 1500 mL for two times (the first time 800 mL; the second 700 mL), at 70℃for 1.5 hours each time. The extracts were combined and the solvent was evaporated to 1 g/mL under pressure. After removing solvent the extract was successively extracted with petroleum ether, ethyl acetate and n-butanol, respectively, and four fractions were got.2 Extraction, isolation and identification: Dried capitulum of Hebei balmy chrysanthemum were extracted two times, with 95% ethanol at 15 times quantity of chrysanthemum (the first 8 times; the second 7 times), for 1.5 hours each time at 70℃. Extracts were combined and the solvent was evaporated to 1 g/mL under pressure. After removing solvent, the extract was successively extracted with petroleum ether, ethyl acetate and n-butanol, respectively. Combined organic layers were evaporated suitably under pressure to give petroleum ether fraction, ethyl acetate fraction and n-butanol fraction. Using silica gel column chromatogram to isolate the remainder of each fractions. Using silica gel column chromatogram, preparation of thin layer chromatogram, polymide column chromatogram, sephadex LH-20 column chromatogram and crystallization, to separate compoundⅠ~Ⅸ. Identification of compounds got from Hebei balmy chrysanthemum were made with UV, 1H-NMR, 13C-NMR, DEPT and MS spectrum.3 Determine total flavonoid: Using macroporous resin column chromatogram, to get 60% ethanol fraction, which has the effect of antihypertension. Using UV to determine flavonoid of this part at 328nm.Result: 1 The preliminary test showed that the extraction percentages of the total extraction, petroleum ether fraction, ethyl acetate fraction and n-butanol fraction Hebei balmy chrysanthemum were 30.0%, 6.4%, 9.2%, 6.1%, respectively.2 The study of chemical composition showed that: nine compounds were isolated from the flowers of Hebei balmy chrysanthemum, and seven of them were identified by spectroscopy methods. They areβ-sitosterol (Ⅱ), acacetin (Ⅳ), 3,5,3'-trihydroxy-6,7,4'-trimethoxy flavone (Ⅴ), chrysoeriol (Ⅵ), apigenin (Ⅶ), acacetin-7-O-β-D-glucoside (Ⅷ), luteolin-7-O-β-D-glucoside (Ⅸ).3 The result of determination showed that there are 37.35% flavonoid in 60% ethanol fraction. Conclusion: The results of our experiment indicated that the solvents and methods of extraction used in this experiment are practicable. With silica gel, polymide and sephadex LH-20 column chromatogram, preparation of thin layer chromatogram and crystallization, the components in Hebei balmy chrysanthemum capitulum were separated and purified, successively. Nine compounds were isolated, and seven of them were identified for the first time in the plant of Hebei balmy chrysanthemum.By macroporous resin column chromatogram, we got 60% ethanol fraction, and using UV to determine flavonoid at 328nm. The result showed that 37.35% flavonoid is in the part of 60% ethanol fraction, and the pharmacology experiments of our group showed that 60% ethanol fraction has the effect of antihypertension.