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中国精品科技期刊2020
旷小珊, 高雄, 林晓蓉, 陈忠正, 李斌, 张媛媛. 南昆山毛叶茶三种多酚的分离纯化及抗氧化研究[J]. 食品工业科技, 2020, 41(5): 31-39. DOI: 10.13386/j.issn1002-0306.2020.05.006
引用本文: 旷小珊, 高雄, 林晓蓉, 陈忠正, 李斌, 张媛媛. 南昆山毛叶茶三种多酚的分离纯化及抗氧化研究[J]. 食品工业科技, 2020, 41(5): 31-39. DOI: 10.13386/j.issn1002-0306.2020.05.006
KUANG Xiao-shan, GAO Xiong, LIN Xiao-rong, CHEN Zhong-zheng, LI Bin, ZHANG Yuan-yuan. Isolation,Purification and Antioxidant Activities of Three Kinds of Polyphenol Monomers from Camellia ptilophylla Chang[J]. Science and Technology of Food Industry, 2020, 41(5): 31-39. DOI: 10.13386/j.issn1002-0306.2020.05.006
Citation: KUANG Xiao-shan, GAO Xiong, LIN Xiao-rong, CHEN Zhong-zheng, LI Bin, ZHANG Yuan-yuan. Isolation,Purification and Antioxidant Activities of Three Kinds of Polyphenol Monomers from Camellia ptilophylla Chang[J]. Science and Technology of Food Industry, 2020, 41(5): 31-39. DOI: 10.13386/j.issn1002-0306.2020.05.006

南昆山毛叶茶三种多酚的分离纯化及抗氧化研究

Isolation,Purification and Antioxidant Activities of Three Kinds of Polyphenol Monomers from Camellia ptilophylla Chang

  • 摘要: 本研究以南昆山毛叶绿茶为材料,通过30%乙醇、乙酸乙酯提取,进而采用羟丙基葡聚糖凝胶(Sephadex LH-20)柱层析和制备型高效液相色谱分离纯化得到3种多酚单体物质;通过高效液相色谱-串联质谱、核磁共振和红外光谱鉴定单体结构;以1,1-二苯基-2-三硝基苯肼(DPPH)和2,2'-联氮基-双-(3-乙基苯并噻唑-6-磺酸)二铵盐(ABTS)两种自由基的清除能力、铁离子还原能力(FRAP)和氧自由基的吸收能力(ORAC)为指标,运用L-O2肝细胞氧化应激模型,综合评价3种多酚单体物质的抗氧化活性。结果:从南昆山毛叶茶中分离得到3种多酚单体物质,分别为没食子儿茶素没食子酸酯(GCG)、1,2,4,6-四没食子酰葡萄糖(1,2,4,6-GA-glc)、儿茶素-3,5-二没食子酸酯(GC-3,5-diGA),其中后2种为茶叶中的特殊多酚物质;纯度分别为98%、96%、97%,得率分别为2.45%、0.55%、0.31%;DPPH、ABTS和FRAP三种化学抗氧化结果基本一致:GCG > GC-3,5-diGA > 1,2,4,6-GA-glc,ORAC法的结果与此相反,1,2,4,6-GA-glc的作用强于GCG和GC-3,5-diGA;高剂量的GCG、1,2,4,6-GA-glc和GC-3,5-diGA预处理后,受氧化应激的L-O2细胞存活率分别提高了26.98%±1.96%、70.87%±3.61%和35.58%±5.37%。结论:通过Sephadex LH-20柱层析和制备型高效液相色谱技术,得到高纯度的GCG、1,2,4,6-GA-glc和GC-3,5-diGA;抗氧化结果显示,GCG具有较强的化学抗氧化能力,1,2,4,6-GA-glc和GC-3,5-diGA具有强细胞抗氧化能力。

     

    Abstract: In this study,Camellia ptilophylla Chang was extracted with 30% ethanol/ethyl acetate,and then three kinds of polyphenol monomers were isolated and purified by Sephadex LH-20 column chromatography and preparative high performance liquid chromatography,and compound structures were identified by high performance liquid chromatography-tandem mass spectrometry,nuclear magnetic resonance and infrared spectroscopy. Antioxidative activity was evaluated by 1,1-diphenyl-2-picrylhydrazyl(DPPH)and 2,2'-azino-bis-(3-ethylbenzothiazole-6-sulfonic acid)diammonium salt(ABTS)radical scavenging ability,iron ion reducing ability(FRAP)and oxygen radical absorbance capacity(ORAC),and L-O2 hepatocyte oxidative stress model. Result:Three kinds of polyphenol monomers were isolated from the Camellia ptilophylla Chang,namely,gallocatechin gallate(GCG)and 1,2,4,6-tetragalloyl glucose(1,2,4,6-GA-glc),gallocatechin-3,5-digallate(GC-3,5-diGA). Purity was 98%,96% and 97%,yield was 2.45%,0.55% and 0.31%,respectively.DPPH,ABTS and FRAP experiments had bacically the same antioxidant results:GCG > GC-3,5-diGA > 1,2,4,6-GA-glc.In contrast,ORAC method had a stronger effect on 1,2,4,6-GA-glc than GCG and GC-3,5-diGA. After pretreatment with high-dose GCG,1,2,4,6-GA-glc and GC-3,5-diGA,the survival rate of L-O2 cells under oxidative stress was increased by 26.98%±1.96%,70.87%±3.61% and 35.58%±5.37%,respectively. In conclusion,to obtain high purity GCG and two kinds of special polyphenols 1,2,4,6-GA-glc and GC-3,5-diGA,Sephadex LH-20 column chromatography and preparative high performance liquid chromatography are used to separate,antioxidant results show that GCG has strong chemical antioxidant capacity,1,2,4,6-GA-glc and GC-3,5-diGA have strong cellular antioxidant capacity.

     

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