高EGCG茶树活性成分提取工艺优化及其茶多酚抗衰老作用的研究
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摘要
以高EGCG含量茶树品种“1005”鲜叶为材料,对鲜叶杀青、茶多酚浸提、茶多酚分离纯化、EGCG纯化工艺及其茶多酚提取物清除自由基能力和动物体内抗衰老的作用进行了研究。
对鲜叶杀青工艺的研究结果表明,EGCG含量从高到低的杀青方式依次是微波杀青11.13%、烘箱杀青9.58%、锅炒杀青9.27%、蒸汽杀青8.44%,微波杀青比蒸汽杀青、锅炒杀青、烘箱杀青提高了2.69%、1.68%、1.55%,因此杀青方式以微波杀青较好。
茶多酚的浸提、分离纯化工艺的研究结果表明,茶多酚最佳浸提条件为浸提溶剂50%乙醇、浸提次数4次、浸提时间50min、浸提温度70℃、浸提体积比为1:50。DM301树脂比D101、H1020树脂具有更好的分离纯化茶多酚性能。DM301树脂具体的分离纯化荼多酚的参数条件是浸提液PH值在2～3之间、吸附解吸流速2mL/min,解吸溶剂80%丙酮。根据以上的浸提、纯化工艺,以高EGCG茶树、铁观音、黄旦鲜叶为原料,得到了三种茶多酚,茶多酚的平均提取率22%,平均纯度95%以上,高EGCG茶树茶多酚提取率分别比铁观音、黄旦高出4%、2%,茶多酚中EGCG含量从高到低分别高EGCG茶树47.07%、铁观音31.55%、黄旦26.08%。高EGCG茶树荼多酚中EGCG含量比铁观音、黄旦高,分别高出15.52%、20.99%。
在EGCG分离纯化研究表明,高EGCG茶树茶多酚和黄旦在流速为0.5mL/min,60%乙醇为洗脱溶剂下以Sephadex LH-20葡聚糖凝胶柱分离纯化EGCG,高EGCG茶树和黄旦茶多酚的EGCG提取率分别为33.3%和16.6%,纯度均为96%～98%。
茶多酚清除自由基能力和动物体内抗衰老研究表明,高EGCG茶树品种茶多酚比普通茶树的茶多酚具有更强的清除自由基能力,比黄旦的茶多酚清除自由基能力高出9.37%。在果蝇的平均寿命延长方面,高EGCG含量茶树茶多酚比黄旦更能延长果蝇的平均寿命,分别提高了14.5%(雄),14.9%(雌),在果蝇体内的SOD值和总抗氧化值方面,高EGCG含量茶树茶多酚比黄SOD值提高了32.9%(雄)、12.1%(雌),总抗氧化能力值提高了44.1%(雄)、28.5%(雌)。不同浸提溶剂提取的茶多酚在延长果蝇寿命上也是有差异的,以乙醇为浸提剂比以沸水为浸提溶剂提取的茶多酚在果蝇平均寿命上提高了6.51%(雄),6.45%(雌),在果蝇体内的SOD值和总抗氧化值方面,乙醇浸提比沸水浸提的茶多酚更能提高果蝇体内SOD值和总抗氧化能力,SOD值提高了9.53%(雄)、9.43%(雌),总抗氧化能力值提高了31.9%(雄)、18.4%(雌)。
The fresh leaves of the tea tree variety "1005" with high EGCG content was used as materials, to carry out the research on the following areas: the deactivation of enzymes of fresh leaves, the extraction of tea polyphenol, the separation and purification of tea polyphenol, the purification of EGCG, the elimination free radical ability of tea polyphenol extraction and the anti-senile function in vivo.
The results of enzyme deactivation of fresh leaves indicates that the EGCG contents (after enzyme deactivation)from high to low in turn are 11.13% by microwave, 9.58% by drying oven, 9.27% by frying in pan, and 8.44% by stream. The best way for enzyme deactivation is using microwave, as it increased EGCG content by 2.69%, 1.68%, and 1.55% compared to using stream, frying in pan and drying oven, respectively.
The results of extraction, the separation and purification of tea polyphenol indicates that the most suitable craft findings to indicate that the best extraction conditions are 50% ethyl alcohol in the solvent, 4 times extraction, 50 mins, 70℃, volume rate 1:50. Compared to D101 and H1020 resin, the DM301 resin is better on the separation and purification of tea polyphenol. The parameters for the separation and purification of tea polyphenol by DM301 are extraction solution PH 2～3, flow speed 2ml/min for adsorption and desorption, 80% acetone in desorption solution. Based on these results, using high EGCG tea tree, Oolong tea and Huangdan as materials, we got three kinds of tea polyphenols, with average extraction rate 22%, and average purity above 95%. The tea polyphenol extraction rate of high EGCG tea tree was 4% and 2% higher than Oolong tea and Huangdan. The EGCG contents in tea polyphenol from high to low in turn are 47.07% in high EGCG tea tree, 31.55% in Oolong tea, 26.08% in Huangdan. EGCG content in tea polyphenol of high EGCG tea tree is 15.52% and 20.99% higher than Oolong tea and Huangdan.
The research on separation and purification of EGCG indicates that, with flow speed 0.5ml/min, 60% ethyl alcohol in elution solution and Sephadex LH-20 glucosan gelatin column, the EGCG extraction rates of tea polyphenol from high EGCG tea tree and Huangdan were 33.3% and 16.6%, with both of the purities between 96%～981
The results of tea polyphenol elimination free radical ability and the animal in vivo anti-senile research indicated that, compared to the tea polyphenol extracted from normal tea tree, tea polypphenol from high EGCG tea tree had stronger ability to eliminate free radicals, 9.37% higher than those in Huangdan. On the extension of fruit fly's lifetime, the tea polyphenol from high EGCG tea tree was able to extend longer lifetime of fruit fly compared to Huangdan, 14.5% higher (male) and 14.9% higher (female). On the aspects of fruit fly in vivo SOD value and total oxidation resistance value, SOD value for tea polyphenol from high EGCG tea tree is 32.9% (male) and 12.1% (female) higher than Huangdan, and total oxidation resistance values were 44.1% (male) and 28.5% (female) higher. Using different extraction solutions, the results were different on the lifetime extension of fruit fly. Compared to using boiled water, extracted tea polyphenol by ethyl alcohol were 6.51% (male) and 6.45% (female) higher on the average of the fruit fly's lifetime extension. On the aspects of fruit fly in vivo's SOD value and total oxidation resistance value, compared to using boiled water, using ethyl alcohol was better on increasing SOD value and oxidation resistance value, SOD value improved 9.53% (male), 9.43% (female), total oxidation resistance values improved 31.9% (male), 18.4% (female).
对鲜叶杀青工艺的研究结果表明,EGCG含量从高到低的杀青方式依次是微波杀青11.13%、烘箱杀青9.58%、锅炒杀青9.27%、蒸汽杀青8.44%,微波杀青比蒸汽杀青、锅炒杀青、烘箱杀青提高了2.69%、1.68%、1.55%,因此杀青方式以微波杀青较好。
茶多酚的浸提、分离纯化工艺的研究结果表明,茶多酚最佳浸提条件为浸提溶剂50%乙醇、浸提次数4次、浸提时间50min、浸提温度70℃、浸提体积比为1:50。DM301树脂比D101、H1020树脂具有更好的分离纯化茶多酚性能。DM301树脂具体的分离纯化荼多酚的参数条件是浸提液PH值在2～3之间、吸附解吸流速2mL/min,解吸溶剂80%丙酮。根据以上的浸提、纯化工艺,以高EGCG茶树、铁观音、黄旦鲜叶为原料,得到了三种茶多酚,茶多酚的平均提取率22%,平均纯度95%以上,高EGCG茶树茶多酚提取率分别比铁观音、黄旦高出4%、2%,茶多酚中EGCG含量从高到低分别高EGCG茶树47.07%、铁观音31.55%、黄旦26.08%。高EGCG茶树荼多酚中EGCG含量比铁观音、黄旦高,分别高出15.52%、20.99%。
在EGCG分离纯化研究表明,高EGCG茶树茶多酚和黄旦在流速为0.5mL/min,60%乙醇为洗脱溶剂下以Sephadex LH-20葡聚糖凝胶柱分离纯化EGCG,高EGCG茶树和黄旦茶多酚的EGCG提取率分别为33.3%和16.6%,纯度均为96%～98%。
茶多酚清除自由基能力和动物体内抗衰老研究表明,高EGCG茶树品种茶多酚比普通茶树的茶多酚具有更强的清除自由基能力,比黄旦的茶多酚清除自由基能力高出9.37%。在果蝇的平均寿命延长方面,高EGCG含量茶树茶多酚比黄旦更能延长果蝇的平均寿命,分别提高了14.5%(雄),14.9%(雌),在果蝇体内的SOD值和总抗氧化值方面,高EGCG含量茶树茶多酚比黄SOD值提高了32.9%(雄)、12.1%(雌),总抗氧化能力值提高了44.1%(雄)、28.5%(雌)。不同浸提溶剂提取的茶多酚在延长果蝇寿命上也是有差异的,以乙醇为浸提剂比以沸水为浸提溶剂提取的茶多酚在果蝇平均寿命上提高了6.51%(雄),6.45%(雌),在果蝇体内的SOD值和总抗氧化值方面,乙醇浸提比沸水浸提的茶多酚更能提高果蝇体内SOD值和总抗氧化能力,SOD值提高了9.53%(雄)、9.43%(雌),总抗氧化能力值提高了31.9%(雄)、18.4%(雌)。
The fresh leaves of the tea tree variety "1005" with high EGCG content was used as materials, to carry out the research on the following areas: the deactivation of enzymes of fresh leaves, the extraction of tea polyphenol, the separation and purification of tea polyphenol, the purification of EGCG, the elimination free radical ability of tea polyphenol extraction and the anti-senile function in vivo.
The results of enzyme deactivation of fresh leaves indicates that the EGCG contents (after enzyme deactivation)from high to low in turn are 11.13% by microwave, 9.58% by drying oven, 9.27% by frying in pan, and 8.44% by stream. The best way for enzyme deactivation is using microwave, as it increased EGCG content by 2.69%, 1.68%, and 1.55% compared to using stream, frying in pan and drying oven, respectively.
The results of extraction, the separation and purification of tea polyphenol indicates that the most suitable craft findings to indicate that the best extraction conditions are 50% ethyl alcohol in the solvent, 4 times extraction, 50 mins, 70℃, volume rate 1:50. Compared to D101 and H1020 resin, the DM301 resin is better on the separation and purification of tea polyphenol. The parameters for the separation and purification of tea polyphenol by DM301 are extraction solution PH 2～3, flow speed 2ml/min for adsorption and desorption, 80% acetone in desorption solution. Based on these results, using high EGCG tea tree, Oolong tea and Huangdan as materials, we got three kinds of tea polyphenols, with average extraction rate 22%, and average purity above 95%. The tea polyphenol extraction rate of high EGCG tea tree was 4% and 2% higher than Oolong tea and Huangdan. The EGCG contents in tea polyphenol from high to low in turn are 47.07% in high EGCG tea tree, 31.55% in Oolong tea, 26.08% in Huangdan. EGCG content in tea polyphenol of high EGCG tea tree is 15.52% and 20.99% higher than Oolong tea and Huangdan.
The research on separation and purification of EGCG indicates that, with flow speed 0.5ml/min, 60% ethyl alcohol in elution solution and Sephadex LH-20 glucosan gelatin column, the EGCG extraction rates of tea polyphenol from high EGCG tea tree and Huangdan were 33.3% and 16.6%, with both of the purities between 96%～981
The results of tea polyphenol elimination free radical ability and the animal in vivo anti-senile research indicated that, compared to the tea polyphenol extracted from normal tea tree, tea polypphenol from high EGCG tea tree had stronger ability to eliminate free radicals, 9.37% higher than those in Huangdan. On the extension of fruit fly's lifetime, the tea polyphenol from high EGCG tea tree was able to extend longer lifetime of fruit fly compared to Huangdan, 14.5% higher (male) and 14.9% higher (female). On the aspects of fruit fly in vivo SOD value and total oxidation resistance value, SOD value for tea polyphenol from high EGCG tea tree is 32.9% (male) and 12.1% (female) higher than Huangdan, and total oxidation resistance values were 44.1% (male) and 28.5% (female) higher. Using different extraction solutions, the results were different on the lifetime extension of fruit fly. Compared to using boiled water, extracted tea polyphenol by ethyl alcohol were 6.51% (male) and 6.45% (female) higher on the average of the fruit fly's lifetime extension. On the aspects of fruit fly in vivo's SOD value and total oxidation resistance value, compared to using boiled water, using ethyl alcohol was better on increasing SOD value and oxidation resistance value, SOD value improved 9.53% (male), 9.43% (female), total oxidation resistance values improved 31.9% (male), 18.4% (female).
引文
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[2]郑金贵.农产品品质学 第一卷[M].厦门大学出版社,2004:338-350.
[3]谭仁祥主编.植物成分功能 第一版[M].科学出版社,2003:217-218.
[4]中国食品添加剂生产应用工业协会编著.食品添加剂手册.第二版[M].中国轻工业出版社,1996,12:46-48.
[5]宛晓春主编.茶叶生物化学 第三版[M].中国农业出版社,2003:9-15.
[6]陆爱霞等.茶多酚提取和应用研究进展[J].食品科技,2002,(2):53-55.
[7]罗晓明.荼多酚浸提工艺研究[J].工艺技术,2002,23(3):41-43
[8]杨贤强等.茶叶中天然抗氧化剂的提取[J].浙江大学学报,1992,18(1):19.
[9]熊何健等.茶多酚分离制备的新工艺[J].食品工业科技,1997,(6):32-34.
[10]廖学品,陆忠兵等.胶原纤维吸附除去茶多酚中的咖啡因[J].离子交换与吸附,2003,19(3):222-228.
[11]姜绍通,潘丽军.醋酸乙酯逆流萃取绿茶茶多酚的研究[J].农业工程学,1997,13(4),202-206.
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