| Zanthoxylum bungeanum belongs to Rutaceae family. It is a traditional medicinal and edible materials in China. Zanthoxylum bungeanum is rich in flavonoids, which has been reported at home or abroad rarely. In this paper, extraction, determination, purification, separation and antioxidation of the flavonoid from Zanthoxylum bungeanum have been studied. The main contents were as follows:(1) The different color react systems to determine flavonoids were compared by the UV-visible light scanning. And the aluminium nitrate-sodium nitrite colorimetry was selected to determine flavonoids of Zanthoxylum bungeanum. The flavonoids of four kinds of Zanthoxylum from different areas were determined, the content of that from Maowen was194.4mg/g, Yunnan189.0mg/g, Hancheng167.4mg/g, Sichuan162.0mg/g. HPLC determination of four kinds of pepper in the content of rutin and quercetin. Rutin content of Zanthoxylum from Hancheng was the highest, up to17.29mg/g. The flavonoid glycosides of total flavonoids hydrolysis was also determined by HPLC. Content of quercetin was the most components, which is up to42.97mg/g.(2) The orthogonal test method was addopted to optimize the extraction technology of total flavonoids from Zanthoxylum bungeanum with ethanol. Based on single-factor test, the orthogonal test was carried out. The results showed that the optimal extraction conditions of total flavonoids were determined to be: ethanol concentration60%, extraction temperature70℃, solid to liquid ratio1:40, and extraction time1h. Under these conditions, the extraction yield of total flavonoids was19.87%.(3) The optimization of purification conditions with macroporous adsorption resin of flavonoids from Zanthoxylum bungeanum were studied in order to improve the total flavonoids content. The optimal techniques of separation and purification of flavonoid were obtained by static adsorption and elution tests, and the separation techniques were evaluated by measuring the concent ration of total flavonoids of Zanthoxylum bungeanum with ultraviolet spectrophotometer. The results showed AB-8macroporous resin showed the best effect of purification. The purification conditions are as follows:sample concent3.0-4.0mg/mL, sample pH4.5, the eluting rate2.5mL/min, the column is washed with distilled water and eluted with about400mL of70%ethanol. Under these conditions the flavonoid content increased from51.46%to90.25%, the yeild rate raised1.75times.(4) The antioxidant effect of the flavonoids from Zanthoxylum bungeanum were studied. by DPPH·test, total reducing test, total antioxidant capacity test and the anti-lipoperoxidation test. The results showed that the Zanthoxylum bungeanum flavonoids had antioxidant effects in different degree. The total flavonoids purified showed stronger antioxidant effect than those crude total flavonoids.(5) The flavonoids from Zanthoxylum bungeanum were sepatared by high-speed counter-current chromatography. The separation was performed using a two-phase solvent system composed of chloroform, methanol, n-butanol and water (4:3:0.5:2, V/V) at a flow rate of2.0ml/min, the rotation speed and the detection wavelength were set at850r/min and254nm, respectively. Then the peak six was obtained by reverse elution. Six flavonoids habe been detected by HPLC. The purities are: peak172.7%, peak267.8%, peak389.6%, peak496.5%, peak594.1%, peak691.3%respectively. The structures were characterized as quercetin3-arabinoside, quercetin7-glucoside, rutin and quercetin3-galactoside through H-NMR seperately. |
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