| In this paper, Cabernet Sauvignon grape leaves collected from Bazhou area in southern Xinjiang were used as the subjects to extract resveratrol through four assistant methods, and the initial structure was identified by infrared spectra after purified,and determination of its antioxidant and antibacterial activity,and prepared resveratrol β- cyclodextrin inclusion compound.. The main experimental results are as follows:By response surface methodology(RSM) to optimize the experimental results obtained, the specific optimization for each method are as follows:⑴The optimum technological conditions for extraction of resveratrol from grape leaves by organic solvent extraction:ethanol concentration 40%, smaterial/ liquid ratio 1:28:, extraction time 4h,,temperature 60℃.Under these conditions, the extraction of resveratrol was 14.4594 mg/g;⑵The optimum technological conditions for extraction of resveratrol from grape leaves by ultrasonic assisted extraction: smaterial/ liquid ratio 1:26(ethanol concentration 40%),temperature 56 ℃,extraction time 59 min,ultrasonic power 280 W. Under these conditions, the extraction of resveratrol was 13.5223 mg/g;16.2593 mg/g;⑶The optimum technological conditions for extraction of resveratrol from grape leaves assisted by microwave method: ethanol concentration 40%, microwave power 640 W, extraction time 76 s, smaterial/ liquid ratio 1:92. Under these conditions, the extraction of resveratrol was 13.5223 mg/g; ⑷ The optimum technological conditions for extraction of resveratrol from grape leaves by enzyme assisted extraction: Enzyme dosage 91 mg(5g enzymatic grape leaves),temperature 52 ℃, time 1.9h, pH5. Under these conditions, the extraction of resveratrol was 16.4523 mg/g。Comparison of four methods shows that extraction rate of microwave-assisted extraction method is lower than other methods, but the extraction time is greatly shortened; extraction rate of enzyme assisted extraction is higher than other methods, but the extraction time is greatly long.By comparing the physical parameters of macroporous resin, static adsorption and desorption rate, resveratrol the crude product was purified using NKA-II resin. The optimum technological conditions were as follows: the sample solution volume is 6 BV,the concentration of resveratrol 3.0 mg/mL, the pH of absorbing stock solution is 4.0, the dynamically absorbing rate of flow is 2 BV/h, With four times the column volume of, 90% ethanol solution as desorption agent under flow rate 3 BV/h.And the results of the four different methods of extraction and purification of resveratrol from grape leaves was studied by infrared spectrum analysis, we found no significant difference in the structure. Through the discovery of infrared spectrum analysis, spectrum characteristic absorption peaks of resveratrol in grape leaves, Description resveratrol grape leaf structure has phenolic hydroxyl, phenyl,-c=c- and other features groups.The four reaction system was studied on the antioxidant activity of resveratrol in grape leaf purification. Experimental results show that, Resveratrol has strong antioxidant activity. Purified resveratrol is directly proportional to its concentration on bacterial inhibition, The purified resveratrol from four extraction methods has no significant difference between the inhibition of E. coli, Microwave assisted extraction of resveratrol from grape leaves were inhibitory to Bacillus subtilis compared to the other three kinds of methods, assisted extraction by organic solvent extraction and microwave grape leaf resveratrol purification product inhibition on Staphylococcus aureus than the other two methods.Through the orthogonal experiment of resveratrol β-cyclodextrin inclusion compound preparation was optimized, depending on the inclusion rate to determine the feed ratio, inclusion temperature and inclusion time, the optimum process is: the ratio of 1:1, inclusion temperature of 50℃, the inclusion time 4h, the resveratrol beta cyclodextrin inclusion rate was 66.0447%. |
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