海南白沙绿茶的抗氧化活性及其酚酸类组成的HPLC-MS分析
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摘要
采用DPPH体系、ABTS体系和Oyaizu法对白沙绿茶的甲醇提取物、70%乙醇提取物和水提取物的抗氧化活性进行测定,并利用HPLC-MS对活性较高的乙醇提取物的抗氧化特征性成分进行分析。结果表明:白沙绿茶3种提取物均具有一定的抗氧化能力,呈浓度效应关系。其中3种提取物中以70%乙醇提取物对DPPH和ABTS清除能力最强,而还原力则弱于水提取物和甲醇提取物。通过HPLC-MS从70%乙醇提取物中得到24个酚类物质,其中15个酚类物质被鉴定为新绿原酸、花旗松素-3-O-芸香糖苷、绿原酸、隐绿原酸、去甲氧基姜黄素、表儿茶素、山奈酚-3-O-芸香糖苷、花旗松素-3-O-葡萄糖苷、槲皮素-双己糖苷、Di-O-咖啡酰奎宁酸、槲皮素、山奈素-3-O-戊糖苷、牛蒡子苷、杨梅素和山奈素。
The antioxidant activity of methanol, 70% ethanol and water extract of Baisha Green Tea was determined by using three complementary methods in vitro: the DPPH radical-scavenging activity, the ABTS radical-scavenging activity, and reducing ability. In addition, the characteristic antioxidant constituents of 70% ethanol extract were analyzed by HPLC-MS. In vitro antioxidant assays showed that the three extracts possessed varying degrees of efficacy in each assay in a dose-dependent manner. 70% ethanol extract exhibited the strongest DPPH radical-scavenging ability and ABTS radical-scavenging ability in the three extracts, while it showed lower reducing ability than water extract and methanol extract. Twenty-four main constituents in the 70% ethanol extract were obtained by HPLC-MS, and the 15 major polyphenols were identified asneochlorogenic acid, taxifolin-3-O-rutinoside,chlorogenic acid, cryptochlorogenic acid, demethoxycurcumin, epicatechin, kampferol-3-O-rutinoside, Taxifolin-3-Ogluconide, quercetin-dihexosides, Di-O-caffeoylquinic acid, kaempferide-3-O-pentoside, arctiin, quercetin, myricetin,and kaempferid.
The antioxidant activity of methanol, 70% ethanol and water extract of Baisha Green Tea was determined by using three complementary methods in vitro: the DPPH radical-scavenging activity, the ABTS radical-scavenging activity, and reducing ability. In addition, the characteristic antioxidant constituents of 70% ethanol extract were analyzed by HPLC-MS. In vitro antioxidant assays showed that the three extracts possessed varying degrees of efficacy in each assay in a dose-dependent manner. 70% ethanol extract exhibited the strongest DPPH radical-scavenging ability and ABTS radical-scavenging ability in the three extracts, while it showed lower reducing ability than water extract and methanol extract. Twenty-four main constituents in the 70% ethanol extract were obtained by HPLC-MS, and the 15 major polyphenols were identified asneochlorogenic acid, taxifolin-3-O-rutinoside,chlorogenic acid, cryptochlorogenic acid, demethoxycurcumin, epicatechin, kampferol-3-O-rutinoside, Taxifolin-3-Ogluconide, quercetin-dihexosides, Di-O-caffeoylquinic acid, kaempferide-3-O-pentoside, arctiin, quercetin, myricetin,and kaempferid.
引文
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[14]海南农垦现代农业工贸股份有限公司.一盏幽兰露五洲共盈香—走向国际的海南名牌—白沙绿茶[J].中国热带农业,2010(1):44-45.
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[22]Hanato T,Kagawa H,Yasuhara T,et al.Two new flavonoids and other constituents in licorice root:their relative astringency and radical scavenging effects[J].Chem Pharm Bull,1998,36:2 090-2 097.
[23]Re R,Pellegrini N,Proteggente A,et al.Antioxidant activity applying an improved ABTS radical cation decolorization assay[J].Free Radical Bio Med,1999,26:1 231-1 237.
[24]Barros L,Baptista P,Ferreira I C F R.Effect of Lactarius piperatus fruiting body maturity stage on antioxidant activity measured by several biochemical assays[J].Food and Chemical Toxicology,2007,45:1 731-1 737.
[25]Oyaizu M.Studies on products of browning reaction:antioxidative activity of products of browning reaction[J].Japanese Journal of Nutrition,1986,40:307-315.
[26]Balentine D A,Wiseman S A,Bouwens L C.The chemistry of tea fiavonoids[J].Critical Reviews in Food Science and Nutrition,1997,37:693-704.
[27]徐春明,乐胜锋,罗晶洁,等.不同种类绿茶提取物抗氧化活性研究[J].中国农学通报,2010,26(16):68-71.
[28]Michalak A.Phenolic compounds and their antioxidant activity in plants growing under heavy metal stress[J].Polish Journal of Environmental Studies,2006,15:523-530.
[29]Velayutham P,Babu A,Liu D.Green tea catechins and cardiovascular health:an update[J].Current Medicinal Chemistry,2008,15:1 840-1 850.
[30]Namal Senanayake S P J.Green tea extract:Chemistry,antioxidant properties and food applications-A review[J].Journal of Functional Foods,2013,5:1 529-1 541.
[31]周金伟,陈雪,易有金,等.不同类型茶叶体外抗氧化能力的比较分析[J].中国食品学报,2014,14(8):262-269.
[32]许红星,曹晖,刘方方.两种方法评价常见水果的抗氧化活性[J].扬州大学烹饪学报,2012(4):38-42.
[33]Surinut P,Kaewsutthi S,Surakarnkul R.Radical scavenging activity in fruit extracts[C]//III WOCMAP congress on medicinal and aromatic plants-volume 5:quality,efficacy,safety,processing and trade in medicinal and aromatic plants.Chiang Mai:Acta Horticulturae,2003:201-203.
[2]魏威.绿茶化学成分及抗氧化性的研究[D].大连:辽宁师范大学,2010.
[3]Abdel-Rahman A,Anyangwe N,Carlacci L,et al.The safety and regulation of natural products used as foods and food ingredients[J].Toxicological Sciences,2011,123:333-348.
[4]Harbowy M E,Balentine D A.Tea chemistry[J].Critical Reviews in Plant Sciences,1997,16(5):415-480.
[5]Higdon J V,Balz F.Tea catechins and polyphenols:Health effects,matabolism,and antioxidant functions[J].Critical Reviews in Food Science and Nutrition,2003,43(1):89-143.
[6]Gordana R,Drazvenka K,Sasva L,et al.Phenolic content and antioxidative capacity of green and white tea extracts depending on extraction conditions and the solvent used[J].Food Chemistry,2008,110:852-858.
[7]An B J,Kwak J H,Son J H,et al.Biological and anti-microbial activity of irradiated green tea polyphenols[J].Food Chemistry,2004,88:549-555.
[8]Macedo J A,Battestin V,Ribeiro M L,et al.Increasing the antioxidant power of tea extracts by biotransformation of polyphenols[J].Food Chemistry,2011,126:491-497.
[9]Khan Naghma,Mukhtar Hasan.Tea polyphenols for health promotion[J].Life Sciences,2007,81(7):519-533.
[10]Kanwar J,Taskeen M,Mohammad I,et al.Recent advances on tea polyphenols[J].Front Biosci(Elite Ed),2012,4:111-131.
[11]侯冬岩,回瑞华,刘晓媛,等.绿茶、红茶和乌龙茶抗氧化性能的比较[J].食品科学,2006,27(3):90-93.
[12]Mondal T K Tea//E C Pua,M R Davey(Eds.).Transgenic Crops V(T Nagata,H Lo··rz,J M Widholm(Eds.).Biotechnology in Agriculture&Forestry,Vol.60)[M].Berlin:Springer,2007:519-535.
[13]Goh R,Gao J,Ananingsih V K,et al.Green tea catechins reduced the glycaemic potential of bread:an in vitro digestibility study[J].Food Chem,2015,180:203-210.
[14]海南农垦现代农业工贸股份有限公司.一盏幽兰露五洲共盈香—走向国际的海南名牌—白沙绿茶[J].中国热带农业,2010(1):44-45.
[15]符耀彩,若石,吴国政.海南白沙绿茶身价倍增[N].农民日报,2004-12-04(第008版).
[16]陈丽珍,翟锐锐,李娟.海南白沙绿茶中维生素C含量的测定[J].广州化工,2012(6):103-104.
[17]谢伟娜,衷明华.毛细管区带电泳法测定白沙绿茶汤中的咖啡因、茶氨酸、EC和EGCG[J].江西化工,2013(2):121-124.
[18]郭安南.绿茶、红茶和咖啡中几种功能成分的分离鉴定及抗氧化性能的比较[J].宁波大学学报(理工版),2011:24(1):18-23.
[19]Jia W,Chu X G,Chang J,et al.High-throughput screening of vitamins and natural antioxidants in nutraceuticals from green tea extracts by liquid chromatography coupled to quadrupole orbitrap mass spectrometry[J].Journal of Chromatography A,2015,1406:337-341.
[20]Singleton V L,Rossi J A Jr.Colorimetric of total phenolics with phosphomolybdic-phosphotungstic acid reagents[J].American Journal of Enology and Viticulture,1965,16:144-158.
[21]罗杨合.Folin-Ciocalteu法测定马蹄皮中多酚的含量[J].湖北农业科学,2009,48(9):2 247-2 249.
[22]Hanato T,Kagawa H,Yasuhara T,et al.Two new flavonoids and other constituents in licorice root:their relative astringency and radical scavenging effects[J].Chem Pharm Bull,1998,36:2 090-2 097.
[23]Re R,Pellegrini N,Proteggente A,et al.Antioxidant activity applying an improved ABTS radical cation decolorization assay[J].Free Radical Bio Med,1999,26:1 231-1 237.
[24]Barros L,Baptista P,Ferreira I C F R.Effect of Lactarius piperatus fruiting body maturity stage on antioxidant activity measured by several biochemical assays[J].Food and Chemical Toxicology,2007,45:1 731-1 737.
[25]Oyaizu M.Studies on products of browning reaction:antioxidative activity of products of browning reaction[J].Japanese Journal of Nutrition,1986,40:307-315.
[26]Balentine D A,Wiseman S A,Bouwens L C.The chemistry of tea fiavonoids[J].Critical Reviews in Food Science and Nutrition,1997,37:693-704.
[27]徐春明,乐胜锋,罗晶洁,等.不同种类绿茶提取物抗氧化活性研究[J].中国农学通报,2010,26(16):68-71.
[28]Michalak A.Phenolic compounds and their antioxidant activity in plants growing under heavy metal stress[J].Polish Journal of Environmental Studies,2006,15:523-530.
[29]Velayutham P,Babu A,Liu D.Green tea catechins and cardiovascular health:an update[J].Current Medicinal Chemistry,2008,15:1 840-1 850.
[30]Namal Senanayake S P J.Green tea extract:Chemistry,antioxidant properties and food applications-A review[J].Journal of Functional Foods,2013,5:1 529-1 541.
[31]周金伟,陈雪,易有金,等.不同类型茶叶体外抗氧化能力的比较分析[J].中国食品学报,2014,14(8):262-269.
[32]许红星,曹晖,刘方方.两种方法评价常见水果的抗氧化活性[J].扬州大学烹饪学报,2012(4):38-42.
[33]Surinut P,Kaewsutthi S,Surakarnkul R.Radical scavenging activity in fruit extracts[C]//III WOCMAP congress on medicinal and aromatic plants-volume 5:quality,efficacy,safety,processing and trade in medicinal and aromatic plants.Chiang Mai:Acta Horticulturae,2003:201-203.