葡萄藤茎中多酚物质提取及其抗氧化能力的研究

葡萄藤茎中多酚物质提取及其抗氧化能力的研究

THE EXTRACTION AND ANTIOXIDANT ACTIVITY OF POLYPHENOLS OF THE GRAPEVINE CANE

作者：孙玉霞　　导师：林建强

山东大学　　生物工程2011届硕士

摘　要

葡萄是多年生藤本植物，每年都有大量修剪下的藤茎被废弃焚烧掉。本文以开发利用葡萄栽培管理过程中的废弃资源为目的，以冬季修剪的葡萄藤茎为试材，优化了多酚物质的提取条件；对其中的多酚类物质进行了定性、定量分析；研究了葡萄藤茎提取液对二苯带苦味酰肼自由基（DPPH）的清除能力。主要包括以下内容：

以总酚含量为考察指标，通过溶剂对比试验和单因素试验确定了最佳提取溶剂及影响多酚提取率的主要因素，并在此基础上采用正交试验法和响应面分析两种试验设计相结合的方法，优化了最佳提取工艺条件：以乙醇为溶剂，提取温度60℃、料液比1:16.5、乙醇溶液浓度50%、提取时间1小时。

据此提取条件测定了2种土壤条件下、5个品种的葡萄藤茎中总酚、总黄酮、单宁、白藜芦醇以及没食子酸、儿茶素、表儿茶素、咖啡酸、阿魏酸和对香豆酸的含量，其中总酚含量13.89mg/g～23.30mg/g,总黄酮量153.93～263.00μg/g,单宁含量7.97～12.69mg/g,白藜芦醇含量15.27～91.63μg/g ，没食子酸13.80～26.19μg/g，其他5种单体酚为全部检出。单宁是含量最高的组分，各品种间多酚类物质的含量存在显著性差异，而且沙砾土中的各组分含量均高于壤土中的。

研究了5个品种葡萄藤茎提取液对DPPH自由基的清除能力，结果表明品种间的差异性显著，且自由基的清除能力与总酚、总黄酮及单宁的含量均呈正相关性。对葡萄藤茎提取液和二丁基羟基甲苯（BHT）及抗坏血酸的自由基清除能力进行了对比研究，葡萄藤茎提取液对DPPH自由基的清除能力远高于相当浓度的BHT对DPPH自由基的清除能力，但较相当浓度的抗坏血酸略低。

关键词　葡萄藤茎　多酚　含量　DPPH　自由基清除能力

Abstract

The aim of this paper is to make use of the grapevine cane waste of viticulture. The grapevine cane pruned in winter was used for qualitative and quantitative analyses of polyphenols. The DPPH free radical scavenging activity of the grapevine cane extracts was also studied. The main conclusions were as follows:

Based on the experimental results of solvent extraction and single factor extraction tests,orthogonal test and response surface analysis were applied to obtain the optimum conditions for extraction of polyphenols. The optimum extraction process of polyphenols from the grapevine cane was as follows: extracting temperature 60℃,ratio of solid to liquid 1:16.5,alcohol concentration 50%,extracting time 1 h .

The content of total phenolic,total flavones ,tannins,t-resveratrol,garlic acid,catechin,epicatechin,caffeic acid,ferulic acid ,p-coumatic acid of 5 Vitis vinifera varieties from 2 soil types were determined. The results indicated that the contents of total phenolic 13.89mg/g～23.30mg/g,total flavones 153.93～263.00μg/g,tannins 7.97～12.69mg/g,resveratrol 15.27～91.63μg/g ， gallic acid 13.80～26.19μg/g，and the content of tannins was the highest of all components. All the samples showed significant difference with varieties. All the content in sand was higher than those in loam.

The DPPH free radical scavenging activity of the grapevine cane extracts of 5 varieties was studied,The results showed that the scavenging activity was in positive correlation with the contents of total phenolics ,total flavones and tannins. The scavenging activity of grapevine cane extract was much higher than BHT of the same concentration,but slightly lower than ascorbic acid.

Key words　Grapevine　Cane　Polyphenols　DPPH　Free radical scavenging activity