黑加仑花色苷的分离纯化及其热降解动力学
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摘要
以黑加仑为原料,采用AB-8大孔树脂-Sephadex LH-20凝胶柱层析联用方法和液相色谱-质谱联用技术对黑加仑花色苷进行分离纯化和组分鉴定;分析了不同纯度花色苷在不同pH和温度下的降解动力学;通过1,1-二苯基-2-三硝基苯肼(DPPH)和2,2'-联氮基-双-(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)自由基的清除评价了不同纯度花色苷的抗氧化能力。结果表明:黑加仑中包含飞燕草素-3-葡萄糖苷、矢车菊素-3-芸香糖苷、牵牛花素-3-葡萄糖苷、芍药素-3,5-二己糖苷和锦葵素-3-半乳糖苷5种组分。经分离纯化后最终获得2种花色苷,分别为飞燕草素-3-葡萄糖苷(A_3)和矢车菊素-3-芸香糖苷(A_4)。pH 3.0和温度50℃时,花色苷的热稳定性最强。不同纯度花色苷组分热降解均符合一级动力学模型。经大孔树脂纯化后的花色苷(A_1)、乙酸乙酯萃取后的花色苷(A_2)、A_3和A_4对DPPH自由基清除率的半数抑制浓度(IC50)分别为9.45、8.17、5.95和7.62 mg/L,而对ABTS自由基清除率的IC50分别为99.38、97.21、78.19和85.54 mg/L。
Anthocyanins of blackcurrant were isolated and purified by combination of AB-8 macroporous resin and Sephadex LH-20, and components were identified by liquid chromatography-mass spectrometry.Thermal degradation kinetics of anthocyanins with different purity was analyzed at different pH values and temperatures. Besides, their antioxidant activities were assessed by measuring their scavenging capacities toward 1,1-diphenyl-2-picrylhydrazyl(DPPH) free radical and 2,2-amino-di(3-ethyl-benzothiazoline sulphonic acid-6) ammonium salt(ABTS) free radical. The results showed that anthocyanins obtained from blackcurrant were mainly delphinidin-3-glucoside, cyanidin-3-rutinoside, petunidin-3-glucoside, paeoniflorin-3,5-dihexoside and malvidin-3-galactoside. The two purified anthocyanins fractions were obtained, which were identified as delphinidin-3-glucoside(A_3) and cyanidin-3-rutinoside(A_4), respectively. The thermal stability of anthocyanins was the strongest at pH 3.0 and temperature of 50 ℃. The thermal degradation of anthocyanins with different purity followed the first order reaction kinetics. The half maximal inhibitory concentration(IC50) of anthocyanins purified by macroporous resin(A_1), anthocyanins extracted by ethyl acetate(A_2), A_3 and A_4 on DPPH free radical were 9.45, 8.17, 5.95 and 7.62 mg/L, respectively. While the IC50 of A_1, A_2, A_3 and A_4 on ABTS free radical were 99.38, 97.21, 78.19 and 85.54 mg/L, respectively.
Anthocyanins of blackcurrant were isolated and purified by combination of AB-8 macroporous resin and Sephadex LH-20, and components were identified by liquid chromatography-mass spectrometry.Thermal degradation kinetics of anthocyanins with different purity was analyzed at different pH values and temperatures. Besides, their antioxidant activities were assessed by measuring their scavenging capacities toward 1,1-diphenyl-2-picrylhydrazyl(DPPH) free radical and 2,2-amino-di(3-ethyl-benzothiazoline sulphonic acid-6) ammonium salt(ABTS) free radical. The results showed that anthocyanins obtained from blackcurrant were mainly delphinidin-3-glucoside, cyanidin-3-rutinoside, petunidin-3-glucoside, paeoniflorin-3,5-dihexoside and malvidin-3-galactoside. The two purified anthocyanins fractions were obtained, which were identified as delphinidin-3-glucoside(A_3) and cyanidin-3-rutinoside(A_4), respectively. The thermal stability of anthocyanins was the strongest at pH 3.0 and temperature of 50 ℃. The thermal degradation of anthocyanins with different purity followed the first order reaction kinetics. The half maximal inhibitory concentration(IC50) of anthocyanins purified by macroporous resin(A_1), anthocyanins extracted by ethyl acetate(A_2), A_3 and A_4 on DPPH free radical were 9.45, 8.17, 5.95 and 7.62 mg/L, respectively. While the IC50 of A_1, A_2, A_3 and A_4 on ABTS free radical were 99.38, 97.21, 78.19 and 85.54 mg/L, respectively.
引文
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[2]Xia Qile(夏其乐),Chen Jianbing(陈剑兵),Gao Haiyan(郜海燕),et al.Effect of sodium D-isoascorbate on the quality of blueberry jam[J].Food Science(食品科学),2018,39(2):20-24.
[3]Rubinskiene M,Jasutiene I,Venskutonis P R,et al.HPLCdetermination of the composition and stability of blackcurrant anthocyanins[J].Journal of Chromatographic Science,2005,43(9):478-482.
[4]Jia Na(贾娜),Kong Baohua(孔保华),Liu Qian(刘骞),et al.Effect of heating temperature and time on the content and antioxidant activity of anthocyanins from black currant[J].Food Science(食品科学),2012,33(21):73-77.
[5]Hellstr?m J,Mattila P,Karjalainen R.Stability of anthocyanins in berry juices stored at different temperatures[J].Journal of Food Composition&Analysis,2013,31(1):12-19.
[6]Dobson G,Mcdougall G J,Stewart D,et al.Effects of juice matrix and pasteurization on stability of black currant anthocyanins during storage[J].Journal of Food Science,2016,82(1):44-52.
[7]Zhao Yuhong(赵玉红),Jia Linna(贾琳娜),Bao Yihong(包怡红),et al.Effect of malic acid on the stability of anthocyanins in blackcurrant juice[J].Modern Food Science and Technology(现代食品科技),2016,32(9):127-134.
[8]Yu Zeyuan(于泽源),Zhao Jianhui(赵剑辉),Li Xingguo(李兴国),et al.Isolation and purification of anthocyanin from blueberry by sequential medium pressure column chromatography on macroporous resin and Sephadex LH-20[J].Food Science(食品科学),2018,39(1):118-123.
[9]Gong Hui(龚辉),Fu Li(傅丽),Li Lanqi(李澜奇),et al.Effect of ultrasound treatment on anthocyanin stability and degradation kinetics[J].Food Science(食品科学),2018,39(9):67-75.
[10]Buchweitz M,Speth M,Kammerer D R,et al.Impact of pectin type on the storage stability of black currant(Ribes nigrum,L.)anthocyanins in pectic model solutions[J].Food Chemistry,2013,139:1168-1178.
[11]Feng Yan(封燕),Gong Xiaohui(贡小辉),Wei Dequn(韦德群),et al.Antioxidant activity and preliminary structure analysis of polysaccharides from cordyceps cicadas[J].Food Science(食品科学),2016,37(13):19-24.
[12]Li Nan(李楠),Shi Junling(师俊玲),Wang Kun(王昆).Composition and in vitro antioxidant activity of polyphenols extracted from crabapples[J].Food Science(食品科学),2014,35(5):53-58.
[13]Cao Shaoqian(曹少谦),Liu Liang(刘亮),Zhang Chao(张超),et al.Purification and thermal degradation kinetics of anthocyanins from mulberry[J].Journal of Chinese Institute of Food Science and Technology(中国食品学报),2015,15(5):54-62.
[14]NicouéE E,Savard S,Belkacemi K.Anthocyanins in wild blueberries of Quebec:Extraction and identification[J].Journal of Agricultural&Food Chemistry,2007,55(14):5626-5635.
[15]Eguchi K,Sato T.Differences in the ratios of cyanidin-3-glucoside and cyanidin-3-rutinocide to total anthocyanin under UV and non-UV conditions in tartary buckwheat(Garten)[J].Plant Production Science,2015,12(2):150-155.
[16]Li D,Meng X,Li B.Profiling of anthocyanins from blueberries produced in China using HPLC-DAD-MS and exploratory analysis by principal component analysis[J].Journal of Food Composition&Analysis,2016,47(6):1-7.
[17]Pertuzatti P B,Barcia M T,Rebello L P G,et al.Antimicrobial activity and differentiation of anthocyanin profiles of rabbiteye and highbush blueberries using HPLC-DAD-ESI-MS,and multivariate analysis[J].Journal of Functional Foods,2016,26:506-516.
[18]Wang Y,Zhu J,Meng X,et al.Comparison of polyphenol,anthocyanin and antioxidant capacity in four varieties of Lonicera caerulea berry extracts[J].Food Chemistry,2016,197(Pt A):522-529.
[19]Chorfa N,Savard S,Belkacemi K.An efficient method for high-purity anthocyanin isomers isolation from wild blueberries and their radical scavenging activity[J].Food Chemistry,2016,197(Pt B):1226-1234.
[20]Zhang Pingping(章萍萍).Study on extraction,purification and activity of antioxidant and prebiotics of purple sweet potato anthocyanins[D].Hefei:Hefei University of Technology(合肥农业大学),2017:3-5.
[21]Jiang Xinlong(蒋新龙).Black rice anthocyanin degradation characteristics[J].Journal of the Chinese Cereals and Oils Association(中国粮油学报),2013,28(4):27-31.
[22]He Chenyang(何晨阳),Wang Zhongbo(王忠博),Fan Chunxue(范春雪),et al.Thermal degradation kinetics of anthocyanins from red cabbage[J].Science and Technology of Food Industry(食品工业科技),2018,39(4):33-43.
[23]Wang Mengze(王梦泽).Study on the stability and quality degradation kinetics of cloudy strawberry juice[D].Yangling:Northwest Agriculture and Forestry University(西北农林科技大学),2010:41-45.
[24]Cao Shaoqian(曹少谦).Studies on structure of anthocyanins from blood orange and related degradation mechanism during processing[D].Wuhan:Huazhong Agricultural University(华中农业大学),2009.
[25]Williams M,Hrazdina G.Anthocyanins as food colorants:Effect of pH on the formation of anthocyanin-rutin complexes[J].Journal of Food Science,2010,44(1):66-68.
[26]Sun Yun(孙芸),Xu Baocai(徐宝财),Gu Wenying(谷文英),et al.Study on relationship between degree of polymerization and radical scavenging capacity of grape seed procyanidins[J].Food Science(食品科学),2007,28(12):423-428.