【作者】 徐洁；

【导师】 胡博然；

【作者基本信息】 扬州大学 ， 食品科学， 2011， 硕士

【摘要】 原花青素是葡萄籽多酚的主要活性物质,现代研究表明,原花青素不仅具有高效的抗氧化活性,而且还有保护心血管、抗癌抗辐射、皮肤保健及美白等功效。本文对葡萄籽中原花青素的提取工艺、分离纯化及抗氧化活性进行以下几个方面的研究：以儿茶素为标准品,采用浓硫酸-香草醛法测定原花青素含量；以原花青素的提取量为指标,通过单因素实验确立影响原花青素提取量的相关因素,并通过正交实验确定提取的最佳条件。对原花青素进行超声波助提实验,筛选到提取的最佳工艺条件为：超声功率80W、料液比1：35、超声时间30min、乙醇浓度50%,原花青素含量为55.728mg/g；对原花青素进行微波助提实验,筛选到提取的最佳工艺条件为：料液比1：25、微波功率115W、微波时间40s、乙醇浓度80%,原花青素含量为106.936mg/g；对原花青素进行酶解助提实验,筛选到提取的最佳工艺条件为：酶解温度45。℃、pH4.0、酶解时间1.0h、酶液浓度100u/mg,原花青素含量为87.513mg/g。通过静态吸附、解吸试验,从5种不同极性的大孔树脂中筛选得到,AB-8树脂的吸附、解吸效果最好,吸附率和解吸率分别达到78.00%、92.42%。选用AB-8树脂对动态吸附条件进行优化实验,确定浓度为3mg/mL的原花青素以3BV/h的流速流经树脂柱时,AB-8的吸附率最高可达到78.19%；对解吸条件进行优化实验,用100mL浓度为30%的乙醇溶液以2BV/h的流速对吸附饱和后的树脂进行洗脱,原花青素的解吸率高达92.37%。将10%-50%乙醇洗脱液进行冻干处理,对原花青素冻干粉的纯度进行测定,得到该混合物的纯度为85.16%。采用四种不同处理样品：原花青素粗提液(GSP0)、25%乙醇洗脱液(GSP1)、50%乙醇洗脱液(GSP2)、50%乙醇洗脱液冻干粉(GSP3),分别进行抗氧化活性实验的研究。1.在清除HO·的实验中,以浓度相同的Vc溶液为对照,检测四种样品的抗氧化性,结果表明：不同样品清除率的大小依次是GSP3、Vc、GSP2、GSP1、GSPO,当浓度大于2.3mg/mL时,Vc溶液的清除率大于GSP3。2.在清除O2·-的实验中,原花青素对02·-的清除率与浓度呈正相关,不同样品清除率的大小依次是：GSP3、GSP2、GSP1、GSP0、Vc。3.在测定还原力的实验中,由700nm下测得吸光值曲线可以看出,还原力与原花青素的浓度呈正比,且它们之间还原力大小关系是：GSP0>GSP1>GSP3>GSP2>Vc.4.在抗油脂氧化能力的实验中,与未添加抗氧化剂的样品相比,原花青素有一定的抗油脂氧化能力,动物油、植物油的过氧化值分别为64.22meq/kg(动物油对照为144.83meq/kg)、433.31meq/kg(植物油对照为573.72meq/kg),且与植物油相比,原花青素对动物油的保护作用更明显。5.利用电子顺磁共振技术测定原花青素提取液对DPPH清除率,不同样品对DPPH清除能率的大小依次是：GSP3、GSP0、GSP2、GSP1,样品中原花青素浓度与清除率呈正相关,与低浓度原花青素溶液相比,高浓度溶液对DPPH的清除具有高效性。更多还原

【Abstract】 Proanthocyanidins are the main active substances of grape seed polyphenols. Modern research shows that, as antioxidants, proanthocyanidins do not only have high activity, But also protect cardiovascular, anti-cancer, anti-radiation, care and white skin. In this experment, extraction technics, purification and antioxidant activity of this substances were studied as follow:Proanthocyanidins content was determined colorimetrically by the vanillin-H2SO4assay using the catechin as the standard. With the content as index, the optimal extraction conditions were determined by the single factor and orthogonal experiment. In the ultrasonic extraction experiments, the results showed that the reaction conditions were ultrasonic power80W, solid-liquid ratio1:35, ultrasonic time30min, concentration of alcohol50%, and the extration of procyanidins content was55.728mg/g. In the microwave extraction experiments, the results showed that the reaction conditions were solid-liquid ratio1:25, microwave power115W, microwave time40s, concentration of alcohol80%, and the extration of procyanidins content was106.936mg/g. In the enzymatic extraction experiment, the results showed that the reaction conditions were temperature45℃, pH4.0, hydrolysis time1.0h, enzyme concentration100u/mg, and the extration of procyanidins content was87.513mg/g.Through the static adsorption experiments,5types of macroporous absorbent resins with different polarities were choosen for the best one. The result showed that an AB-8resin had a better adsorption (78.00%) and desorption (92.42%) properties than four other resins examined. The best adsorption condition was found to be3mg/mL procyanidins concentration with a flow rate of3BV/h, and the final adsorption rate was78.19%. The optimal desorption condition was found to be a flow rate of2BV/h using100mL of30%ethanol, and the final desorption rate of proanthocyanidins was92.37%. With the method of freeze-dried,10%-50%ethanol and other impurity of the mixture were removed, and the purity of the proanthocyanidins were determined to be85.16%.The samples of studying the antioxidant activity were different. They were crude extracts (GSPO), procyanidins solution mixed with25%ethanol (GSP1),50%ethanol (GSP2) and procyanidins powder after freeze-dried (GSP3).1. Procyanidins could effectively scavenge OH compared with Vc. The scavenging activity of four different samples was GSP3, Vc, GSP2, GSP1, GSPO, and when the concentration of procyanidins overed2.3mg/mL, the scavenging activity of Vc was stronger than GSP3.2. The O2·-radical-scavenging activity had a regative correlation with the concentration of procyanidins. And the scavenging activity of four different samples was GSP3, GSP2, GSP1, GSPO, Vc.3. The result of reducing power experment showed that the reducing power had a regative correlation with the concentration of procyanidins, and the relationship between them was GSP0> GSP1> GSP3> GSP2> Vc.4. Compared with the sample not added oil, procyanidins could inhibit peroxidation of lard and lipid. The POV of animal oil was64.22meq/kg (the control was144.83meq/kg), and the POV of vegetable oil was433.31meq/kg (control was573.72meq/kg). And the effect of protecting animal oil from peroxidating was much better than the vegetable oil.5. In the experment of DPPH scavenging effect of proanthocyanidins by ESR, the scavenging ability of them was GSP3, GSPO, GSP2, GSP1. The scavenging effect had a regative correlation with the concentration of procyanidins, and compared with low concentrations of procyanidins, the other was more efficient on DPPH scavenging effect.更多还原