Extraction, Isolation And Antioxidant Activity Of Flavonoids From Acanthopanax Sessiliflorus Fruit

Acanthopanax sessiliflorum has been named as WuYazi, not only can be used for people to watch, but also is a good crop for medicinal. It’s also a kind of nutritious vegetables in northeast area of China. In recent years, people aways ues it instead of Acanthopanax for medicinal.The existence of flavonoid is universal in the biological world with a variety of functions such as elimination of free radicals, anti-oxidation, anti-cancer, lowering blood sugar, inhibiting activity, against cancer and so on. The content of flavonoid in Acanthopanax sessiliflorum fruit is very rich, which is associated with a variety of medicinal value. This test shows the result of overall extraction, purification, analysis, identification of the flavonoids in Acanthopanax sessiliflorum fruit. And the antioxidant properties were in vitro studies. The results are as follows:1-, Based on the results of single factor analysis, we make the response surface to optimize parameter on extraction process of total flavonoids in Acanthopanax sessiliflorum fruit. Combined with the actual experimental operating conditions, we ultimately determine the optimum operating parameters as follows:extracting for 40 minutes of 30%ethanol with liquid ratio of 1:50. The test was repeated three times in parallel, the actual content of the extraction is (54.2124 ± 0.3458) mg/g, which reached 99.71% of the expected value.2, In order to purified the initial flavonoid further, we go on with macroporous. Through the comparative analysis of static adsorption and desorption tests, we choose resin type HPD-600 with the highest adsorption rate of (52.06 ± 0.033)%, and the higher desorption rate of (73.98 ± 0.032)%. In order to optimize the process conditions of HPD-600 macroporous resin, six aspect of test was conducted. Ultimately the determination of optimal conditions for the purification process is that:the flow rate of adsorption solution was 1.0mL/min, the volume was 120mL, the concentration 0.6mg/mL, the flow rate of desorption solution was 1.OmL/min, the volume fraction was 1 OOmL with 70% ethanol.3、Dissolving the flavonoids of Acanthopanax sessiliflorum fruit purified by resin in water. Separate the flavonoid components further with ethyl acetate. The measurement results showed that the flavonoids are mainly concentrated in the ethyl acetate layer of (173.96± 1.32) mg/g, far higher than the content of (36.79± 0.72) mg/g in the aqueous layer. And the flavonoids in aqueous layer mainly containing the anthocyanin of C5 position hydroxyl group. The anthocyanin wasn’t included in the ethyl acetate layer. Comparison analysis with a standard composition of the main components of flavonoid by HPLC shows the ingredient were rutin, Hyperoside, quercetin. By LC-MS technology to further analyze its composition, the results show that it also contains protocatechuic, new chlorogenic acid, Dicaffeylquinic acid. With the comparation of the characteristic ion fragment in charge to mass ratio, the anthocyanins substances in aqueous layer may contain mallow pigments.4、 By measuring the purified flavonoids of Acanthopanax sessiliflorum fruit, blueberries, apples on DPPH radical scavenging, superoxide anion radical scavenging, hydroxyl radical scavenging, ABTS radical scavenging activity, the test shows the antioxidant study of Acanthopanax sessiliflorum fruit flavonoids. The results showed that the antioxidant activity of flavonoids in Acanthopanax sessiliflorum fruit is the best, which DPPH radical scavenging IC50 value was 27.44μg/mL, the IC50 value of superoxide anion radical scavenging was 24.21 μg/mL, the IC50 value of hydroxyl free radical scavenging was 10.49μg/mL, the IC50 values of ABTS radical scavenging was (5.26±0.13) mmol/g.