Fertilization And Flavonoids Of The Flos Lonicerae Japonicae

Flos Lonicerae Japonicae are the drying buds of Lonicera japonica Thunb., a perennial evergreen winding shub of Caprifoliaceae. Flos Lonicerae Japonicae is used in Chinese herbal medicine for latent-heat-clearing and detoxicant. Both modern chemical analyzing and pharmaceutical experienments convinced that Flos Lonicerae Japonicae had the functions of anti-fungal and antimicrobial activities, anti-virus, anti-tumour, detoxicant, immunosuppressive, anti-inflammatory, liver protection, homeostasis, anti-ulceration and so on. Because of its special pharmaceutical effects, Flos Lonicerae Japonicae had played an important role in the period of"SARS". As a result, Flos Lonicerae Japonicae had become popular in the Chinese herbal medicine researches.In order to exploit and utilize this plant resource deeply, this paper investigated the fertilization, the antifungal activities and the flavonoids of the Flos Lonicerae Japonicae. Through studying on the effects of fertilization on the quality of Chinese honeysuckle, the optimal fertilization formula was acquired. The antifungal activities were investigated. Based on the theories and principles of phytochemistry and modern chemical separation and purification methods, three different extracts from different organic solvents of Flos Lonicerae Japonicae's antifungal activities were studied and the flavonoids contents were studied by silica gel and polyamide chromatography. The main results obtained from the experiments are as follows:1. The quality regulat on the Flos Lonicerae JaponicaeThree different proportions of N and P fertilizer in different amounts of fertilizer were used to investigate the effects of fertilization on the quality of Chinese honeysuckle, which is located in GAP Base of Honeysuckle in Nanzheng, Shaanxi. The results showed that the optimal amount is 560 kg·hm-2 with the proportion of N:P2O5 1:0.78,Which could increase the yield of honerysuckle by 103% than the control. The chlorogenic acid content and the total flavonoids content were increased by 22.13% and 23.25%, respectively.2. The antifungal activities of the Flos Lonicerae Japonicae The petroleum ether extract, ethanol extract and water extract of the Flos Lonicerae Japonicae had antifungal activities at different degree. Water extract had the highest antifungal activity. The follow one was ethanol extract. And their inhibitory effect was concentration dependent. The petroleum ether extract's antifungal activity was the worst, and even had proving effects on the growth of the phytopathol fungi at low concentration in this experiment. Water extract had inhibitory effects on 5 types phytopathol fungi at all three levels, and the inhibition on Fusarium ulbigenum was the best. The highest inhibition rate was 63.75%. Compared with water extract, ethanol extract had weaker antifungal activities, even could improve the growth of some fungi at low concentration. And ethanol extract was the most effective to the Alternaria solani. The inhibition rate was as high as 50.00%. Petroleum ether extract had antifungal activities at high concentration. But when the concentration was reduced to 0.1mg·ml-1, it showed strong improving effects on the growth of fungi in the experiment.3. The extraction of total flavonoids from Flos Lonicerae Japonicae by ultrasonicThe total flavonoids were extracted from Flos Lonicerae Japonicae using alcohol by ultrasonic. Taking four factors, the alcohol concentration, the ultrasonic extracting time, the solid-liquid ratio of the Flos Lonicerae Japonicae to alcohol and the extracting temperature into account, the yield of the total flavanoids from Flos Lonicerae Japonicae was investigated in the experiment. The results showed that the optimum extracting conditions were: The ultrasonic extracting time was 25 minutes, alcohol concentration was 70 %, the solid-liquid ratio of the Flos Lonicerae Japonicae to alcohol was 1:30, and the extracting temperature was 50℃. The extraction rate was as high as 7.85%.4. The separation of flavonoids in the alcohol extract of Flos Lonicerae JaponicaeThe separation and purification of flavonoids in the alcohol extract of Flos Lonicerae Japonicae were carried out by using thin layer chromatography, column chromatography and recrystallization. Four compounds were separated from the ethanol extract of Flos Lonicerae Japonicae.Two of them were flavonoids. Their structures were identified basing on their H-NMR,13C-NMR,MS spectrum data, melting points and compared Rf value with the standards. They were identified asβ-sitosterol(II), daucosterol, Luteolin-7-O-β-D-glucoside(Ⅲ),rutin(Ⅳ).5. The purification of Luteolin-7-O-β-D-glucoside from Flos Lonicerae JaponicaeLuteolin-7-O-β-D-glucoside was purified from the alcohol extract of Flos Lonicerae Japonicae within two times through polyamide column chromatography. Alcohol extract of Flos Lonicerae Japonicae was subjected to polyamide column chromatography using alcohol-water gradient elution. The 30-40% alcohol-water fraction was eluted with CHCl3–MeOH gradient elution by through polyamide column. Luteolin-7-O-β-D-glucoside was obtained in CHCl3–MeOH (15/1), with the purity more than 98%, the yield more than 8.78%. This method overcame the disadvantage of flavonoids tailing on the silica gel column chromatography and was easy to operate for large scale preparation of Luteolin-7-O-β-D-glucoside. It also can separated chlorogenic acid at the same time.Flos Lonicerae Japonicae has been used for Chinese traditional medicine for thousands of years. It was popular for its efficiencies on the diseases and its gentle character. This paper studied the fungistatic activities and flavonoids of the Flos Lonicerae Japonicae. The results regarded that Flos Lonicerae Japonicae can be used as the plant resource of the pesticide. Two flavonoids were obtained from the ethanol extract of Flos Lonicerae Japonicae. Furthermore, this study sets up an easy way to obtain Luteolin-7-O-β-D-glucoside. These results enriched the research contents of Flos Lonicerae Japonicae and offered the theory basis for rational exploitation and utilization.