朝鲜蓟叶多酚提取及抗氧化活性研究
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摘要
本研究主要通过单因素和正交试验,探讨了朝鲜蓟叶多酚的最佳提取工艺,比较了大孔树脂纯化前后朝鲜蓟叶多酚提取物的抗氧化和抑菌活性的差异。结果表明,最佳提取工艺为:乙醇浓度70%,料液比1:20,超声时间40min,该条件下多酚提取量为88.36mg/g。通过9种大孔树脂静态吸附及解吸性能的比较,确定了NKA-2型树脂为朝鲜蓟多酚的最佳纯化树脂。经过大孔树脂纯化后,朝鲜蓟叶多酚的抗氧化能力显著增强,如对DPPH、ABTS+自由基清除的IC50值分别从82.79μg/m L和251.30μg/m L,降低到32.30±0.25μg/m L、107.98±0.42μg/m L。另外,纯化工艺还显著提高了朝鲜蓟叶提取物的抑菌效果,对金黄色葡萄球菌、大肠杆菌、枯草芽孢杆菌、白色念珠菌、绿脓杆菌和乳杆菌均有抑制作用。综上所述,该研究为朝鲜蓟叶多酚的提取、纯化和应用提供了科学依据和理论基础。
In this study, the optimal conditions for extracting polyphenols from artichoke leaves were investigated using single factor and orthogonal tests. The antioxidant and antibacterial activities of the polyphenol extracts from artichoke leaves before and after the purification by macroporous resins were compared. The results showed the optimal extraction conditions were: extraction solvent, 70% ethanol; sample/solvent ratio, 1:20(g/mL); ultrasonic time, 40 min. Under these conditions, the total phenol content of the extract from artichoke leaves was 88.36 mg/g.Based on the comparison of static adsorption and desorption of nine macroporous resins, NKA-2 resin was found to be the best option for purification of the polyphenol extract. After such purification, the antioxidant activity of the purified polyphenol extract from artichoke leaves was significantly increased e.g. the IC50 values of 1-diphenyl-2-trinitrobenzene hydrazine(DPPH) and 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) radical scavenging decreased to 32.30±0.25 μg/m L and 107.98 ± 0.42 μg/m L from 82.79 μg/m L and 251.30 μg/m L,respectively. In addition, the purification process significantly improved the antibacterial effect of the extract from artichoke leaves, including the inhibition of Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Monilia albican, Pseudomonas aeruginosa and Lactobacillus. In summary, this work provides a scientific basis and theoretical foundation for the extraction, purification and application of the polyphenols from artichoke leaf.
In this study, the optimal conditions for extracting polyphenols from artichoke leaves were investigated using single factor and orthogonal tests. The antioxidant and antibacterial activities of the polyphenol extracts from artichoke leaves before and after the purification by macroporous resins were compared. The results showed the optimal extraction conditions were: extraction solvent, 70% ethanol; sample/solvent ratio, 1:20(g/mL); ultrasonic time, 40 min. Under these conditions, the total phenol content of the extract from artichoke leaves was 88.36 mg/g.Based on the comparison of static adsorption and desorption of nine macroporous resins, NKA-2 resin was found to be the best option for purification of the polyphenol extract. After such purification, the antioxidant activity of the purified polyphenol extract from artichoke leaves was significantly increased e.g. the IC50 values of 1-diphenyl-2-trinitrobenzene hydrazine(DPPH) and 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) radical scavenging decreased to 32.30±0.25 μg/m L and 107.98 ± 0.42 μg/m L from 82.79 μg/m L and 251.30 μg/m L,respectively. In addition, the purification process significantly improved the antibacterial effect of the extract from artichoke leaves, including the inhibition of Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Monilia albican, Pseudomonas aeruginosa and Lactobacillus. In summary, this work provides a scientific basis and theoretical foundation for the extraction, purification and application of the polyphenols from artichoke leaf.
引文
[1]Gebhardt R.Antioxidative and protective properties of extracts from leaves of the artichoke(Cynara scolymus L.)against hydroperoxide-induced oxidative stress in cultured rat hepatocytes[J].Toxicology&Applied Pharmacology,1997,144(2):279-286
[2]Kraft K.Artichoke leaf extract-Recent findings reflecting effects on lipid metabolism,liver and gastrointestinal tracts[J].Phytomedicine International Journal of Phytotherapy&Phytopharmacology,1997,4(4):369-378
[3]Aktay G,Deliorman D,Ergun E,et al.Hepatoprotective effects of Turkish folk remedies on experimental liver injury[J].Journal of Ethnopharmacology,2000,73(1):121-129
[4]Wang M F,Li J G,Rangarajan M,et al.Antioxidative phenolic compounds from sage(Salvia officinalis)[J].Journal of Natural Products,1999,62(3):454-456
[5]Gaafar A A,Salama Z A,Baz F K E.Antioxidant and antiproliferative effects on human liver Hep G2 epithelial cells from artichoke(Cynara scolymus L.)by-products[J].Journal of Natural Sciences Research,2013,3(10):17-24
[6]Saénz R T,García G D,De I P V R.Choleretic activity and biliary elimination of lipids and bile acids induced by an artichoke leaf extract in rats[J].Phytomedicine,2002,9(8):687-693
[7]Zhu X F,Zhang H X,Lo R,et al.Antimicrobial activities of Cynara scolymus L.leaf,head,and stem extracts[J].Journal of Food Science,2010,70(2):M149-M152
[8]Liu S W,Chang X Dg,Liu X F,et al.Effects of pretreatments on anthocyanin composition,phenolics contents and antioxidant capacities during fermentation of hawthorn(Crataegus pinnatifida)drink[J].Food Chemistry,2016,212:87-95
[9]李柏榆,杨美莲,曹建新,等.超声辅助提取桑葚多酚及抗氧化活性分析[J].云南民族大学学报(自然科学版),2017,26(2):97-102LI Bo-yu,YANG Mei-lian,CAO Jian-xin,et al.Extraction and antioxidant activity of total phenolic compounds from mulberry[J].Journal of Yunnan Nationalities University:Natural Sciences Edition,2017,26(2):97-102
[10]Ruan X,Zhan L M,Gao X X,et al.Separation and purification of flavonoid from Taxus remainder extracts free of taxoids using polystyrene and polyamide resin[J].Journal of Separation Science,2013,36(12):1925-1934
[11]刘清,李虎虎,姚惠源,等.大孔吸附树脂分离富集大麦多酚的研究[J].食品科学,2007,28(11):98-102LIU Qing,LI Hu-hu,YAO Hui-yuan,et al.Study on isolation and enrichment of barley polyphenols by macroporous resin[J].Food Science,2007,28(11):98-102
[12]Mensor L L,Menezes F S,Leit?o G G,et al.Screening of Brazilian plant extracts for antioxidant activity by the use of DPPH free radical method[J].Phytotherapy Research,2001,15(2):127-130
[13]Zheng X L,Znang R T,Liu C.Extraction and antioxidant activity of phenolic compounds from wheat bran treated by steam explosion[J].Tropical Journal of Pharmaceutical Research,2015,14(10):1857-1863
[14]Sun D,Huang S Q,Cai S B,et al.Digestion property and synergistic effect on biological activity of purple rice(Oryza sativa L)anthocyanins subjected to a simulated gastrointestinal digestion in vitro[J].Food Research International,2015,78(2):114-123
[15]高凡丁,黄士淇,张成庭,等.发酵青刺果种子的酚类物质组成及抗氧化活性分析[J].云南民族大学学报(自然科学版),2018,27(3):167-175GAO Fan-ding,HUANG Shi-qi,ZHANG Cheng-ting,et al.Analysis of phenolic composition and antioxidant activities in fermented Prinsepia utilis Royle seeds[J].Journal of Yunnan Nationalities University:Natural Sciences Edition,2018,27(3):167-175
[16]Floegel A,Kim D O,Chung S J,et al.Comparison of ABTS/DPPH assays to measure antioxidant capacity in popular antioxidant-rich US foods[J].Journal of Food Composition&Analysis,2011,24(7):1043-1048
[17]付慧,杨萍,汪秋宽.茶多酚的制备及其抑菌活性的研究[J].食品科技,2012,37(6):273-276FU Hui,YANG Ping,WANG Qiu-kuan.Preparation and antimicrobial activities of tea polyphenols[J].Food Science and Technology,2012,37(6):273-276
[18]陈建中,葛水莲,昝立峰,等.响应面试验优化双水相萃取大吴风草总黄酮工艺及抑菌活性测定[J].食品科学,2015,36(24):57-62CHEN Jian-zhong,GE Shui-lian,ZAN Li-feng,et al.Optimization of aqueous two-phase extraction of total flavonoids from Farfugium japonicum(L.f.)Kitam by response surface methodology and antibacterial activity[J].Food Science,2015,36(24):57-62
[19]杨克沙.朝鲜蓟的化学成分和活性研究[D].昆明:昆明理工大学,2015YANG Ke-sha.Study on the chemical constituents and bioactivity of Cynara scolymus[D].Kunming:Kunming University of science and Technology,2015
[20]张明,帅希祥,杜丽清,等.澳洲坚果青皮多酚提取工艺优化及其抗氧化活性[J].食品工业科技,2017,38(21):195-199.ZHANG Ming,SHUAI Xi-xiang,DU Li-qing,et al.Optimization of extraction and antioxidant activity of polyphenols from macadamia green peel[J].Science and Technology of Food Industry,2017,38(21):195-199
[21]旷慧,迟超,吕长山,等.红树莓多酚的醇法提取工艺优化[J].食品科学,2016,37(10):88-93KUANG Hui,CHI Chao,LYU Chang-shan,ea al.Optimization of ethanol extraction process for polyphenols from red raspberry[J].Food Science,2016,37(10):88-93
[22]苗修港,余翔,张贝贝,等.NKA-9大孔树脂纯化香椿叶黄酮类物质工艺优化[J].食品科学,2016,27(8):32-38MIAO Xiu-gang,YU Xiang,ZHANG Bei-bei,et al.Purification of flavonoids from Toona sinensis leaves with NKA-9 macroporous resin[J].Food Science,2016,27(8):32-38
[23]Zan M A,Ferraz A B F,Richter M F,et al.In vivo,genotoxicity evaluation of an Artichoke(Cynara scolymus L.),aqueous extract[J].Journal of Food Science,2013,78(2):T367-371
[24]赵艳红,李建科,李国荣.石榴皮多酚纯化及其抗氧化活性表征[J].食品科学,2010,31(11):31-37ZHAO Yan-hong,LI Jian-ke,LI Guo-rong.Purification with macroporous adsorbent resins and in vitro antioxidant evaluation of pomegranate peel polyphenols[J].Food Science,2010,31(11):31-37
[25]蒋孟君,王艺,任建青,等.超声提取食用玫瑰花总酚及其大孔树脂纯化前后抗氧化活性[J].食品工业科技,2017,38(23):164-169JIANG Meng-jun,WANG Yi,REN Jian-qing,et al.Study on ultrasonic extraction of total phenol from edible rose and antioxidant activity of macroporous resin before and after purification[J].Science and Technology of Food Industry,2017,38(23):164-169
[26]Zan M A,Ferraz A B F,Richter M F,et al.In vivo genotoxicity evaluation of an artichoke(Cynara scolymus L.)aqueous extract[J].Journal of Food Science,2013,78(2):5
[27]Wang M F,Simon J E,Aviles I F,et al.Analysis of antioxidative phenolic compounds in artichoke(Cynara scolymus L.)[J].Journal of Agricultural and Food Chemistry,2003,51(3):601-608
[28]Azzini E,Bugianesi R,Romano F,et al.Absorption and metabolism of bioactive molecules after oral consumption of cooked edible heads of Cynara scolymus L.(cultivar Violetto di Provenza)in human subjects:A pilot study[J].British Journal of Nutrition,2007,97(5):963
[29]霍超,宋曙辉,徐桂花,等.朝鲜蓟叶提取物的抑菌作用研究[J].食品科技,2009,34(9):217-219HUO Chao,SONG Shu-hui,XU Gui-hua,et al.Study on antimicrobial effects of artichoke leaves extract[J].Food Science and Technology,2009,34(9):217-219
[30]李芬芳,马艳弘,赵密珍,等.草莓多酚的提取工艺优化及其抑菌活性研究[J].江苏农业科学,2017,45(3):155-158LI Fen-fang,MA Yan-hong,ZHAO Mi-zhen,et al.Optimization of extraction process of strawberry polyphenols and its antimicrobial activity evaluation[J].Jiangsu Agricultural Sciences,2017,45(3):155-158
[31]Zhu X,Zhang H,Lo R.Phenolic compounds from the leaf extract of artichoke(Cynara scolymus L.)and their antimicrobial activities[J].Journal of Agricultural and Food Chemistry,2004,52(24):7272-7278
[32]王建超.枇杷叶多酚提取、纯化及其抗氧化、抑菌活性研究[D].福州:福建农林大学,2015WANG Jian-chao.Study on extraction and purification process,antioxidant and antibacterial activity of polyphenols from loquat leaves[D].Fuzhou:Fujian Agriculture and Forestry University,2015
[2]Kraft K.Artichoke leaf extract-Recent findings reflecting effects on lipid metabolism,liver and gastrointestinal tracts[J].Phytomedicine International Journal of Phytotherapy&Phytopharmacology,1997,4(4):369-378
[3]Aktay G,Deliorman D,Ergun E,et al.Hepatoprotective effects of Turkish folk remedies on experimental liver injury[J].Journal of Ethnopharmacology,2000,73(1):121-129
[4]Wang M F,Li J G,Rangarajan M,et al.Antioxidative phenolic compounds from sage(Salvia officinalis)[J].Journal of Natural Products,1999,62(3):454-456
[5]Gaafar A A,Salama Z A,Baz F K E.Antioxidant and antiproliferative effects on human liver Hep G2 epithelial cells from artichoke(Cynara scolymus L.)by-products[J].Journal of Natural Sciences Research,2013,3(10):17-24
[6]Saénz R T,García G D,De I P V R.Choleretic activity and biliary elimination of lipids and bile acids induced by an artichoke leaf extract in rats[J].Phytomedicine,2002,9(8):687-693
[7]Zhu X F,Zhang H X,Lo R,et al.Antimicrobial activities of Cynara scolymus L.leaf,head,and stem extracts[J].Journal of Food Science,2010,70(2):M149-M152
[8]Liu S W,Chang X Dg,Liu X F,et al.Effects of pretreatments on anthocyanin composition,phenolics contents and antioxidant capacities during fermentation of hawthorn(Crataegus pinnatifida)drink[J].Food Chemistry,2016,212:87-95
[9]李柏榆,杨美莲,曹建新,等.超声辅助提取桑葚多酚及抗氧化活性分析[J].云南民族大学学报(自然科学版),2017,26(2):97-102LI Bo-yu,YANG Mei-lian,CAO Jian-xin,et al.Extraction and antioxidant activity of total phenolic compounds from mulberry[J].Journal of Yunnan Nationalities University:Natural Sciences Edition,2017,26(2):97-102
[10]Ruan X,Zhan L M,Gao X X,et al.Separation and purification of flavonoid from Taxus remainder extracts free of taxoids using polystyrene and polyamide resin[J].Journal of Separation Science,2013,36(12):1925-1934
[11]刘清,李虎虎,姚惠源,等.大孔吸附树脂分离富集大麦多酚的研究[J].食品科学,2007,28(11):98-102LIU Qing,LI Hu-hu,YAO Hui-yuan,et al.Study on isolation and enrichment of barley polyphenols by macroporous resin[J].Food Science,2007,28(11):98-102
[12]Mensor L L,Menezes F S,Leit?o G G,et al.Screening of Brazilian plant extracts for antioxidant activity by the use of DPPH free radical method[J].Phytotherapy Research,2001,15(2):127-130
[13]Zheng X L,Znang R T,Liu C.Extraction and antioxidant activity of phenolic compounds from wheat bran treated by steam explosion[J].Tropical Journal of Pharmaceutical Research,2015,14(10):1857-1863
[14]Sun D,Huang S Q,Cai S B,et al.Digestion property and synergistic effect on biological activity of purple rice(Oryza sativa L)anthocyanins subjected to a simulated gastrointestinal digestion in vitro[J].Food Research International,2015,78(2):114-123
[15]高凡丁,黄士淇,张成庭,等.发酵青刺果种子的酚类物质组成及抗氧化活性分析[J].云南民族大学学报(自然科学版),2018,27(3):167-175GAO Fan-ding,HUANG Shi-qi,ZHANG Cheng-ting,et al.Analysis of phenolic composition and antioxidant activities in fermented Prinsepia utilis Royle seeds[J].Journal of Yunnan Nationalities University:Natural Sciences Edition,2018,27(3):167-175
[16]Floegel A,Kim D O,Chung S J,et al.Comparison of ABTS/DPPH assays to measure antioxidant capacity in popular antioxidant-rich US foods[J].Journal of Food Composition&Analysis,2011,24(7):1043-1048
[17]付慧,杨萍,汪秋宽.茶多酚的制备及其抑菌活性的研究[J].食品科技,2012,37(6):273-276FU Hui,YANG Ping,WANG Qiu-kuan.Preparation and antimicrobial activities of tea polyphenols[J].Food Science and Technology,2012,37(6):273-276
[18]陈建中,葛水莲,昝立峰,等.响应面试验优化双水相萃取大吴风草总黄酮工艺及抑菌活性测定[J].食品科学,2015,36(24):57-62CHEN Jian-zhong,GE Shui-lian,ZAN Li-feng,et al.Optimization of aqueous two-phase extraction of total flavonoids from Farfugium japonicum(L.f.)Kitam by response surface methodology and antibacterial activity[J].Food Science,2015,36(24):57-62
[19]杨克沙.朝鲜蓟的化学成分和活性研究[D].昆明:昆明理工大学,2015YANG Ke-sha.Study on the chemical constituents and bioactivity of Cynara scolymus[D].Kunming:Kunming University of science and Technology,2015
[20]张明,帅希祥,杜丽清,等.澳洲坚果青皮多酚提取工艺优化及其抗氧化活性[J].食品工业科技,2017,38(21):195-199.ZHANG Ming,SHUAI Xi-xiang,DU Li-qing,et al.Optimization of extraction and antioxidant activity of polyphenols from macadamia green peel[J].Science and Technology of Food Industry,2017,38(21):195-199
[21]旷慧,迟超,吕长山,等.红树莓多酚的醇法提取工艺优化[J].食品科学,2016,37(10):88-93KUANG Hui,CHI Chao,LYU Chang-shan,ea al.Optimization of ethanol extraction process for polyphenols from red raspberry[J].Food Science,2016,37(10):88-93
[22]苗修港,余翔,张贝贝,等.NKA-9大孔树脂纯化香椿叶黄酮类物质工艺优化[J].食品科学,2016,27(8):32-38MIAO Xiu-gang,YU Xiang,ZHANG Bei-bei,et al.Purification of flavonoids from Toona sinensis leaves with NKA-9 macroporous resin[J].Food Science,2016,27(8):32-38
[23]Zan M A,Ferraz A B F,Richter M F,et al.In vivo,genotoxicity evaluation of an Artichoke(Cynara scolymus L.),aqueous extract[J].Journal of Food Science,2013,78(2):T367-371
[24]赵艳红,李建科,李国荣.石榴皮多酚纯化及其抗氧化活性表征[J].食品科学,2010,31(11):31-37ZHAO Yan-hong,LI Jian-ke,LI Guo-rong.Purification with macroporous adsorbent resins and in vitro antioxidant evaluation of pomegranate peel polyphenols[J].Food Science,2010,31(11):31-37
[25]蒋孟君,王艺,任建青,等.超声提取食用玫瑰花总酚及其大孔树脂纯化前后抗氧化活性[J].食品工业科技,2017,38(23):164-169JIANG Meng-jun,WANG Yi,REN Jian-qing,et al.Study on ultrasonic extraction of total phenol from edible rose and antioxidant activity of macroporous resin before and after purification[J].Science and Technology of Food Industry,2017,38(23):164-169
[26]Zan M A,Ferraz A B F,Richter M F,et al.In vivo genotoxicity evaluation of an artichoke(Cynara scolymus L.)aqueous extract[J].Journal of Food Science,2013,78(2):5
[27]Wang M F,Simon J E,Aviles I F,et al.Analysis of antioxidative phenolic compounds in artichoke(Cynara scolymus L.)[J].Journal of Agricultural and Food Chemistry,2003,51(3):601-608
[28]Azzini E,Bugianesi R,Romano F,et al.Absorption and metabolism of bioactive molecules after oral consumption of cooked edible heads of Cynara scolymus L.(cultivar Violetto di Provenza)in human subjects:A pilot study[J].British Journal of Nutrition,2007,97(5):963
[29]霍超,宋曙辉,徐桂花,等.朝鲜蓟叶提取物的抑菌作用研究[J].食品科技,2009,34(9):217-219HUO Chao,SONG Shu-hui,XU Gui-hua,et al.Study on antimicrobial effects of artichoke leaves extract[J].Food Science and Technology,2009,34(9):217-219
[30]李芬芳,马艳弘,赵密珍,等.草莓多酚的提取工艺优化及其抑菌活性研究[J].江苏农业科学,2017,45(3):155-158LI Fen-fang,MA Yan-hong,ZHAO Mi-zhen,et al.Optimization of extraction process of strawberry polyphenols and its antimicrobial activity evaluation[J].Jiangsu Agricultural Sciences,2017,45(3):155-158
[31]Zhu X,Zhang H,Lo R.Phenolic compounds from the leaf extract of artichoke(Cynara scolymus L.)and their antimicrobial activities[J].Journal of Agricultural and Food Chemistry,2004,52(24):7272-7278
[32]王建超.枇杷叶多酚提取、纯化及其抗氧化、抑菌活性研究[D].福州:福建农林大学,2015WANG Jian-chao.Study on extraction and purification process,antioxidant and antibacterial activity of polyphenols from loquat leaves[D].Fuzhou:Fujian Agriculture and Forestry University,2015