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Extraction, Purification and Effect of Scavenging Free Radicals of Trans-resveratrol from Grape Pomace

Author: ZhuYiWei
Tutor: ZhangXiuLing
School: Northeast Agricultural University
Course: Agricultural Products Processing and Storage
Keywords: Grape pomace Trans-resveratrol An organic solvent Macroporous resin Alkali extraction and acid precipitation Free radicals
CLC: TS261.9
Type: Master's thesis
Year: 2008
Downloads: 305
Quote: 0
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Abstract


In this experiment, after the by-product in the production of wine - grape pomace as raw material extracted from a variety of physiological functions of trans-resveratrol. Conditions of trans-resveratrol extraction and purification methods, and free radical scavenging capacity, to provide a theoretical basis for the development and utilization of grape pomace. Trans-resveratrol detection method using ultraviolet spectrophotometry and high performance liquid chromatography. UV spectrophotometry to determine the optimum absorption wavelength of 306nm, and draw a standard curve regression equation: y = 0.1122x-0.0132 (R2 = 0.9991), the concentration range of 0 to 8μg/mL; high performance liquid chromatography, the chromatographic conditions: mobile phase was methanol - aqueous solution (50:50, v / v), a flow rate of 0.8mL/min, injection volume was 10μL, the column temperature was room temperature, the resulting trans-resveratrol standard chromatogram peak shape better, and the results of the standard curve regression equation: y = 60297x-10418 (R2 = 0.9997). Trans-resveratrol concentration showed a good linear relationship in the range of 0.5μg/mL ~ 20μg/mL, the detection limit was 0.5μg/mL. Grape pomace with 80% methanol, 80% ethanol, 80% acetone and ethyl acetate and other organic solvent extraction was carried out, the results show that the effect of the 80% ethanol extraction of trans-resveratrol. Process to optimize the extraction conditions, the use of the three factors and three levels of response surface methodology. The result of optimum extraction conditions: temperature of 69 ° C for 61min, liquid to solid ratio of 46 mL / g, the theoretical maximum extraction content 104.39μg / g the actual proven experiments extract content 103.86μg / g. The impact of various factors in descending order of priority for liquid-solid ratio gt; extraction time gt; extraction temperature. Macroporous resin method and alkali extraction and acid precipitation and purification process of trans-resveratrol. Macroporous resin method selected AB-8 resin separation and purification, the basic completion of the adsorption of trans-resveratrol around 4h; when the concentration of trans-resveratrol 70.6μg/mL, AB -8 resin adsorption effect; adsorption on the AB-8 resin using 75% ethanol solution, flow rate 0.75 BV / h desorption approximately about the consumption 5.5BV is desorbent can trans-resveratrol basic alcohol desorbed; the trans resveratrol First purity of 26.32%. Alkali extraction and acid precipitation Finally, pH = 12 NaOH solution was extracted with HCl solution adjusted to the pH of the solution = 3. The final product qualitative detection to determine which contains trans-resveratrol, and after HPLC determination, come to a purity of 59.88%. Trans-resveratrol hydroxyl radical and DPPH radical were studied, the results showed that: trans-resveratrol extract on free radical scavenging effect, and the clear ability of the strength depends on the trans-white concentration of alcohol Veratryl. Within the concentration range used in the present experiment, the greater the concentration, the stronger scavenging effect.

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