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Study on Extraction, Isolation and Antioxidant Activity of Resveratrol from Peanut Vine

Author: WangCaiXia
Tutor: WuXiangYang
School: Jiangsu University
Course: Food Science and Engineering
Keywords: Peanut vine Resveratrol HPLC analysis Extraction Process Aqueous two-phase Purification Antioxidant
CLC: R285
Type: Master's thesis
Year: 2009
Downloads: 166
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Abstract

Resveratrol (Resveratrol, Res), also known as Di trihydroxybenzene chemical called trans -3,4 ', 5 - hydroxy -1,2 - stilbene (3,4' ,5-trihydrolystilbence) widely exist in natural plants such as knotweed, grapes (skins, seeds), mulberries, peanuts, pineapple plants for resistance to pathogen infection polyphenols have antioxidant, lowering blood pressure, prevention of cardiovascular vascular diseases, as well as anti-cancer and other important biological activity. The production of resveratrol extract functional foods popular in the international market, the market demand. Currently abroad, mainly from the grape and grape seed extract, domestic mainly from traditional Chinese medicine Polygonum cuspidatum extract, but are facing a shortage of raw material resources, the cost of the problem. Peanut roots, stems and leaves are reportedly contains resveratrol content than grape skin (seed) in the high tens of times to hundreds of times, but the peanut vine industrial scale use, only partly used as animal feed, and the rest discarded as waste, or burning in the field, it would be a waste of agricultural resources and cause environmental pollution. Therefore, the research and development of peanut vines (including peanut roots, stems, leaves) of resveratrol in the expansion of the plant resources and enhance the economic value of the peanut byproducts of resveratrol are of great significance. In this paper, peanut vine as raw materials, the analysis on the the peanut vine in resveratrol extraction process, Purification and antioxidant activity of research, and to provide a basis for the development and utilization of the peanut vine. The main contents are as follows: the peanut vine resveratrol high performance liquid chromatography (HPLC) analysis method. Peanut vine extract impurities, especially the fat-soluble impurities column contamination seriously affect the life of the column, the establishment of peanut root, stem, leaf step by silica gel column pretreatment, HPLC method for the determination of which resveratrol content method. Pretreatment eluted with chloroform: acetone: ethanol: water (4:4:0.5:0.2, V / V / V / V), the TLC method tracking identified, collected the resveratrol components by HPLC analysis. Chromatographic conditions: column Hypersil ODS-2 (4.6 mm × 250 mm, 5μm), the mobile phase was methanol: water (45:55, v / V), the flow rate was 0.8mL/min, column temperature was 35 ° C, UV The detect wavelength of 306nm. The regression equation: Y = 9 × 10 ^{6 X-340 693, linear range :0.228-1 .14 μg (correlation coefficient R = 0.9999). Silica gel column after step pretreatment addition to resveratrol peak in HPLC detection chromatogram few impurity peaks, pretreatment purifying effect. The method is simple, fast, and accurate. Measured from Shandong, Jiangsu and Hebei the peanut vine mixture resveratrol content were 903.69μg / g, 116.21μg / g and 88.69μg / g; Shandong Province peanut vines content of the highest, higher than that reported in the literature. Peanut vine in different parts of Shandong Province peanut roots, stems, leaves resveratrol content measured results were 223.08μg / g, 1211.19μg / g and 429.79μg / g, the highest on the content of the stem. Research the peanut vine in resveratrol optimization of solvent extraction process. Optimized technology of extracting peanut vine reported, mostly by UV spectrophotometric method for the determination of the content of resveratrol. Found at a wavelength of 306nm at peanut vine extract samples 4min before a major impurity peak of HPLC detection impurity peak area is much higher than the peak area of ??resveratrol, such as ultraviolet spectrophotometry is not an accurate reflection Chenopodium quinoa the resveratrol extraction This article as index traditional solvent extraction method to extract resveratrol resveratrol extraction, HPLC method for the determination of the amount of extraction of resveratrol extract samples in the univariate Based on the experiments, the orthogonal experiment, the influences of temperature, ethanol concentration and liquid feed than the effect of resveratrol extract. Temperature is the significant factors affecting the amount of resveratrol extract, solvent extraction, optimization of process parameters: temperature 50 ℃, 60% ethanol, liquid to solid ratio 25:1, extraction time 1h, extraction times, measured under this condition, the extraction amount of resveratrol is 855.05μg / g raw material. The 3 validation tests show that the process is stable and can be used for the extraction of peanut vines in resveratrol. Resveratrol theoretical study of aqueous two-phase extraction system. Have been reported in the literature using aqueous two-phase system of purification of resveratrol from Polygonum cuspidatum, but no aqueous two-phase extraction system in the theoretical study reported resveratrol This article as index partition coefficient and recovery, using ultraviolet spectrophotometry spectrophotometry to determine the content to determine the extraction of resveratrol aqueous two-phase system for ethanol - ammonium sulfate - water and cloud point titration to establish the solubility phase diagrams of aqueous two-phase system, while affecting aqueous two-phase Extraction System resveratrol effect factors were investigated. The results show that the mass fraction of ethanol and ammonium sulfate is the main factor to affect the aqueous two-phase extraction. Create aqueous two-phase extraction of resveratrol in peanut vines. After aqueous two-phase extraction system, desalination anti-extraction process, resveratrol is concentrated in the upper extract and the recovery rate was 97.5%, HPLC method for the measurement of sample resveratrol content of 1.83%, with crude extracts matter content of resveratrol (0.2%) compared to more than nine times. Visible, the purification process of aqueous two-phase system is not only to avoid the use of toxic reagents such as ethyl acetate, and to improve the purity of the samples can be used for preliminary purification of peanut vine Resveratrol. Investigation peanut vine resveratrol crude extract, the aqueous two-phase purified sample and the content of 96.93% resveratrol antioxidant activity of the sample, and compared with V _C. Determined by spectrophotometry sample of 2,2 - diphenyl-1 - bitter the hydrazino radical (DPPH ·), hydroxyl radical (· OH) and superoxide anion radical (O ₂ · -) scavenging effect. The results showed that the free radical scavenging ability of the sample and the content of 96.93% resveratrol sample in the the peanut vine resveratrol crude extracts, aqueous two-phase purified enhanced with the increase of the concentration of the sample, the sample concentration is less than 2.664mg/mL peanut vine Resveratrol crude extract better than free radical scavenging aqueous two-phase purified samples, but not as much content as 96.93% resveratrol samples; the sample concentration 2.664mg/mL, three samples radical scavenging the ability of consistency. The peanut vines resveratrol crude extract the Clear DPPH ·, · OH and O ₂ · - radical IC ₅₀ values ??were 0.0529mg/mL, 0.589mg/mL and 0.610mg/mL; purified by aqueous two-phase the sample clearing DPPH · · OH and O ₂ · - radicals IC50 values ??were 0.0625 mg / mL, 1.684 mg / mL and 2.469mg / mL; the resveratrol Samples clear content of 96.93% DPPH ·, · OH and O ₂ · - the free radicals IC ₅₀ values ??were 0.00624mg/mL, 0.485mg/mL and 0.200mg/mL; V _C the Clear DPPH ·, · OH and O ₂ · - radical IC _{the 50} values were 0.009 mg / mL, 0.255mg/mL 0.190mg/mL, visible free radical scavenging capacity of the three samples were slightly weaker than Vc; three samples of DPPH ·, · OH and O _{2 · scavenging order consistent: the content of 96.93% resveratrol samples gt; the the peanut vine resveratrol crude extracts gt; purified by aqueous two-phase sample; content of 96.93% resveratrol samples free radical scavenging effect Vc scavenging effect quite clear the similar effect Vc radical scavenging effect the peanut vine resveratrol crude extract; aqueous two-phase purified samples free radical scavenging capacity of some weaker.}}

Study on Extraction, Isolation and Antioxidant Activity of Resveratrol from Peanut Vine

Abstract

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