The Extraction, Purification And Identify Of Flavonoids From Bamboo Willows

Bamboo willow as a new species of bamboo and willow, has many characteristics.They were widely used in industrial production. It has very high economic value. But now,the research on the active ingredients in bamboo willow hasn't been found. It was found that bamboo willow contains flavonoids. Flavonoids have a variety of physiological activity,such as antioxidation, antitumor, antibacterial, reducing blood fat, prevent cardiovascular disease. In this study, bamboo willows as the research object, the purification method and extraction process conditions were researched. With LC-MS, HSCCC, IR and NMR technology, the flavonoids were analyzed and identified which have provided the certain experimental basis for deep research and positive significance for the comprehensive utilization of flavonoids from bamboo willow leaves. The main research contents and experimental results were listed respectively as follows:1.The ethanol refluxing extraction process of flavonoids from bamboo willows was used. A series of single-factor experiments were made to investigate the effects of extraction temperature, liquid-solid ration,ethanol concentration and extraction time on the yield of flavonoids, based on these,orthogonal experiment was employed to optimize the extraction conditions of flavonoids. The optimal extraction conditions of flavonoids were extraction temperature of 80?, material-liquid ratio of 1:40(g/mL), ethanol concentration of 50% and extraction time of 2h. Under the optimal conditions, the maximum yield of flavonoids was 4.0%.2.Five kinds of macroporous resins including D101, D4020, GDX201, HPD80 and AB-8 were selected to compare their adsorption rates and elution rates of bamboo willow flavonoids. The optimal macroporous resins AB-8 was decided. The optimal purification conditions of flavonoids by AB-8 resin were sample concentration of 1.0mg/mL, sample flow rate of 1.0mL/min, ethanol concentration of 70%, elution velocity of 0.5ml/min. After treatment with AB-8 resin in purification conditions, the purity of bamboo willow flavonoids can reach the height of 30.9%, which is about 2 times higher than the previous of 11.8%.3.The LCMS-IT-TOF method was applied to analyze flavonoids from bamboo willows.Six flavonoids were speculated with MS/MS2 spectrun. Compared with flavonoids references, five flavonoids including isovitexin-7-O-glucoside, rutin, luteolin-7-O-glucoside, isorhamnetin-3-O-rutinoside, apigenin-7-O-glucuronide were identified. High speed counter current chromatography(HSCCC) was successfully applied to the separation and preparation of compound VI using solvent system composed of n-butyl alcohol-ethyl acetate-methyl alcohol-H2O(1.5:4:1:7). Compounds VI was 92.5% determined by HPLC.With the IR and NMR technology, the sixth flavonoid was speculated as chrysoeriol-glucuronide.