柠檬桉树脂总黄酮的提取纯化及抗氧化活性
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摘要
采用溶剂法提取纯化柠檬桉树脂总黄酮,对粗提物(A)和纯化物进行抗氧化活性研究。用盐酸-锌粉、浓硫酸、三氯化铁、硝酸铝等方法定性鉴别树脂总黄酮,以芦丁为标准品定量分析树脂总黄酮含量为4.64%(以柠檬桉树脂固体粉末的质量为基准,下同)。粗提物(A)经氯仿-乙酸丁酯、氯仿-二甲基甲酰胺、二甲基甲酰胺-氯仿两次超声萃取后得到纯化物B、C、D中总黄酮质量分数分别为10.39%、18.03%、44.15%。考察了A、B、C、D对2,2-二苯基-1-苦味基肼自由基(DPPH·)的清除活性、总抗氧化能力和还原能力。结果表明,质量浓度为0.14 g/L时,A、B、C、D对DPPH·的清除活性分别为91.75%、74.22%、88.44%、91.56%;总抗氧化能力分别为19.99、1.75、4.39、3.79μmol芦丁当量(RE)/g;还原能力分别为1.16、0.36、0.39、0.45μmol RE/g。
Total flavonoids in Eucalyptus citriodiraresin were extracted and purified by solvent method and antioxidant activities of crude extract(A) and purified compounds were studied. The total flavonoids in resin were qualitatively identified by hydrochloric acid-zinc powder, concentrated sulfuric acid, ferric chloride and aluminum nitrate.Total flavonoids content(based on the quality of Eucalyptus citriodira resin solid powder, the same below) was 4.64% by quantitative analysis with rutin as standard substance.The purified compounds B, C and D were obtained by ultrasonic extraction of the crude extract(A) using chloroform and butyl acetate, chloroform and dimethyl formamide, dimethyl formamide and chloroform as extract solvent. Their corresponding mass fraction of total flavonoids were 10.39%, 18.03% and 44.15%.The scavenging activity, total antioxidant capacity and reducing power of A, B, C and D against 2,2-diphenyl-1-bitter hydrazine radical(DPPH) were investigated. When mass concentration was 0.14 g/L,The scavenging activity of A, B, C and D on DPPH· were 91.75%, 74.22%, 88.44% and 91.56%,respectively. The total antioxidant capacity was 19.99, 1.75, 4.39 and 3.79 μmol rutin equivalent(RE)/g.The reducing power was 1.16, 0.36, 0.39 and 0.45 μmol RE/g.
Total flavonoids in Eucalyptus citriodiraresin were extracted and purified by solvent method and antioxidant activities of crude extract(A) and purified compounds were studied. The total flavonoids in resin were qualitatively identified by hydrochloric acid-zinc powder, concentrated sulfuric acid, ferric chloride and aluminum nitrate.Total flavonoids content(based on the quality of Eucalyptus citriodira resin solid powder, the same below) was 4.64% by quantitative analysis with rutin as standard substance.The purified compounds B, C and D were obtained by ultrasonic extraction of the crude extract(A) using chloroform and butyl acetate, chloroform and dimethyl formamide, dimethyl formamide and chloroform as extract solvent. Their corresponding mass fraction of total flavonoids were 10.39%, 18.03% and 44.15%.The scavenging activity, total antioxidant capacity and reducing power of A, B, C and D against 2,2-diphenyl-1-bitter hydrazine radical(DPPH) were investigated. When mass concentration was 0.14 g/L,The scavenging activity of A, B, C and D on DPPH· were 91.75%, 74.22%, 88.44% and 91.56%,respectively. The total antioxidant capacity was 19.99, 1.75, 4.39 and 3.79 μmol rutin equivalent(RE)/g.The reducing power was 1.16, 0.36, 0.39 and 0.45 μmol RE/g.
引文
[1]Editorial Board of《Chinese Materia Medica》of State Administration of Traditional Chinese Medicine of the People’s Republic of China.Chinese materiamedica[M].Shanghai:Shanghai Scientific&Technical Publishers(上海科学技术出版社),1999.
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[9]Zhao Erlao(赵二劳),Yang Jie(杨洁),Zhao Sanhu(赵三虎).Research progress on extraction and purification technologies of flavonoids from peanut hull[J].Journal of The Chinese Cereals and Oils Association(中国粮油学报),2018,33(5):128-134.
[10]Rahul K L,Vaqar M A,Hasan M.Dietary flavonoid fisetin for cancer prevention and treatment[J].Molecular Nutrition&Food Research,2016,60(6):1401-1405.
[11]Du W J,Yang X L,Song Z J.Antitumor activity of total flavonoids from daphne genkwa in colorectal cancer[J].Phytotherapy Research,2015,30(2):323-330.
[12]Xu F,Wang C X,Wang H F,et al.Antimicrobial action of flavonoids from Sedum aizoon L.against lactic acid bacteria in vitro and in refrigerated fresh meat[J].Journal of Functional Foods,2018,40:744-750.
[13]Maria C Si,Laura N C,Franco M E,et al.Potent inhibition of western equine encephalitis virus by a fraction rich in flavonoids and phenolic acids obtained from achyrocline satureioides[J].Revista Brasileira de Farmacognosia,2016,26(5):571-578.
[14]de Souza P Oa,Bianch-Ib S E,Figueiróa F,et al.Anticancer activity of flavonoids isolated from achyrocline satureioides in gliomas cell lines[J].Toxicology in Vitro,2018,51:23-33.
[15]Zhao H,Song L,Huang W.Total flavonoids of epimedium reduce ageing related oxidative DNA damage in testis of rats via p53-depedent pathway[J].Andrologia,2017,49(10):1-7.
[16]Keti Z,Ketan R,Randolph R A,et al.Flavonoids and their metabolites:Prevention in cardiovascular diseases and diabetes[J].Diseases,2017,5(3):9.
[17]Jiang J,Xiao S C,Xua X M,et al.Isomeric flavonoid aglycones derivedfrom epimedii folium exerted different intensities in antiosteoporosis through OPG/RANKL protein targets[J].International Immunopharmacology,2018,62:277-286.
[18]Yi Y S.Regulatory roles of flavonoids on inflammasome activation during inflammatory responses[J].Molecular Nutrition&Food Research,2018,62(13):1-13.
[19]Yan T,Wu B,Liao Z Z,et al.Brainderived neurotrophic factor signaling mediates the a-ntidepressant like efect of the total favonoids of alpiniaeoxyphyllae fructus in chronic unpredictable mild stress mice[J].Phytotherapy Research,2016,30(9):1493-1502.
[20]Huang Liansha(黄链莎),Liu Tonghua(刘铜华),Sun Wen(孙文),et al.Effect and mechanism of flavonoids from mori folium on blood glucose level in diabetic[J].Chinese Journal of Experimental Traditional Medical Formulae(中国实验方剂学杂志),2018,24(16):152-156.
[21]Liang Q,Chen H G,Zhou X,et al.Optimized microwave-assistant extraction combined ultrasonic pretreatment of flavonoids from periplocaforrestiischltr.and evaluation of its antiallergic activity[J].Electrophoresis,2017,38(8):1113-1121.
[22]Lu Xiaoxiang(鲁晓翔).Research progress in antioxidant mechanism of flavonoids[J].Food Research and Development(食品研究与开发),2012,33(3):220-224.
[23]Yu Fuxiang(余付香),Chen Mingxu(陈明旭),Cheng Qiao(成桥),et al.Classified extraction and activity of total flavonoids from penthorum chinense purse[J].Nat Prod Res Dev(天然产物研究与开发).2017,29:976-982.
[24]Cai Jinyuan(蔡锦源),Wei Kunhua(韦坤华),Xiong Jianwen(熊建文),et al.Extraction of flavonoids from Sophora tonkinensis Gapnep and their antioxidant and antibacterial activities[J].Fine Chemicals(精细化工),2017,34(3):285-293.
[25]Wang Shasha(王沙沙),Liu Yang(刘洋),Niu Zhenzhen(牛真真),et al.Separation and purification of total flavonoids from salsolacollina with macroporous resins and evaluation of antioxidant activities[J].Fine Chemicals(精细化工),2018,35(1):58-64.
[26]Wang Fangfang(王芳芳).Studies on chemical constituents of dracaena cochinchinensis and their bioactivities[D].Kunming:Yunnan University of Nationalities(云南民族大学),2015.
[27]He Ying(何颖).The analysis of methods for extracting essential oils[J].Tianjin Pharmacy(天津药学),2015,27(1):47-50.
[28]Yao Xinsheng(姚新生).Natural pharmaceutical chemistry[M].Beijing:People’s Publishing House(人民出版社),1996:194.
[29]Ku Yongfeng(库咏峰),Huang Pinxian(黄品鲜),Chen Ping(陈萍),et al.Study on extrac-tion and separation analysis&antioxidant activity of total flavonoids from cinnamon[J].Application of Chemical(应用化工),2011,40(9):1547-1552.
[30]Zhou Yiming(周一鸣).Study on extraction purification and antioxidative effects of flavoniod sintatary bulk wheat bran[D].Xi'an:Shaanxi Normal University(陕西师范大学),2008.
[31]Zhou Ningbo(周宁波),Li Yujie(李玉杰),Yang Chengxiong(杨成雄),et al.Pharmaceutical analysis[M].Beijing:Chemical Industry Press(化学工业出版社),2010,7.
[32]Zeng Su(曾苏),Chai Yifeng(柴逸峰),Song Fenyun(宋粉云),et al.Pharmaceutical analysis[M].Beijing:Higher Education Press(高等教育出版社),2014:2.
[33]Xing Yun(邢云),Wang Guifang(王桂芳),Li Quanmin(李全民).Trends of solvent extraction in analytical chemistry[J].Physical and Chemical Testing-Chemical Section(理化检验-化学分册),2005,41(9):694-696.
[34]Wang P F,Mu X P,Du J J,et al.Flavonoid content and radical scavenging activity in fruits of Chinese dwarf cherry(C-erasus humilis)genotypes[J].Journal of Forestry Research,2018,29(1):55-63.
[35]Niveditha V R,Sridhar K R.Antioxidant activity of raw,cooked and Rhizopus oligosporus fermented beans of Canavalia of coastal sand dunes of Southwest India[J].Journal of Food Science and Technology,2014,51(11):3253-3260.
[36]Feng C Y,Su S,Wang L J,et al.Antioxidant capacities and anthocyanin ch-aracteristics of the black-red wild berries obt-ained in Northeast China[J].Food Chemistry,2016,204:150-158.
[37]Jeony S C,Jeony Y T,Yang B K,et al.White button mushroom(agaricusbisporus)lowers blood glucose and cholesterol levels in diabetic and hypercholesterolemic rats[J].Nutr Res,2010,30(1):49-56.
[38]Zhong Qiuping(钟秋平),Lin Meifang(林美芳).Determination on of contents of flavones and identifiction of flavones species in fructus clausenaelanii[J].Food Science(食品科学),2007,28(8):411-413.
[39]Feng Yabin(冯雅斌),Du Liang(杜靓),Wen Jing(温静).Study progress about drugs analysis on TLC[J].Disease Monitor&Control Jan(疾病监测与控制杂志),2011,5(1):60-63.
[40]Tian Qinglai(田庆来),Xie Yuchun(谢渝春),Zhang Bo(张波),et al.Separation of hydrophilic licorice flavonoid by solvent extraction method[J].The Chinese Journal of Process Engineering(过程工程学报),2007,7(3):496-500.
[41]Ni Xiaolei(倪晓蕾).Selective liquid-liquid extraction of natural phenolic compounds using ionic liquids[D].Hangzhou:Zhejiang University(浙江大学),2012.
[42]Marina P,Olga L T.Use of different spices as potential natural antioxidant additives on cooked beans(phaseol-us vulgaris).increase of DPPH radical scavenging activity and total phenolic content[J].Plant Foods for Human Nutrition,2014,69(4):337-343.
[43]Mariola S,Joanna N,Jerzy W,et al.Total antioxidant capacity of honey bee haemolymph in relation to age and exposure to pesticide,and comparison to antioxidant capacity of seminal plasma[J].Apidologie,2016,47(2):227-236.
[44]Narumol P,Weerasak S,Patcharawee N,et al.In vitro antioxidant activity study of novel chromone deriv-atives[J].Chemical Biology&Drug Design,2012,79(6):981-989.
[2]Qu Lian(屈恋),Zhang Wenyang(张闻扬),Liu Xiongmin(刘雄民),et al.Analysis and comparison of volatile constituents in the fruits a-nd leaves of Eucalyptus citriodora[J].Science and Technology of Food Industry(食品工业科技),2016,37(12):71-75,88.
[3]Tian Yuhong(田玉红),Liu Xiongmin(刘雄民),Zhou Yonghong(周永红),et al.Extraction and determination of volatile constituents in leaves of Eucalyptus citriodora[J].Chinese Journal of Chromatography(色谱),2005,23(6):651-654.
[4]Lin Sanqing(林三清),Zhou Zongliu(周中流),Yang Hongyan(杨红艳),et al.Extraction techniques and free radical scavenging activity o-f total flavonoids from the leaves of Eucalyptus citriodora[J].China Pharmaceuticals(中国药业),2017,26(6):5-7.
[5]Qu Lian(屈恋),Gao Xuan(高旋),Liu Xiongmin(刘雄民),et al.Study on purifiction and antioxidant activities of tannins of Eucalyptus citriodora fruits[J].Science and Technology of Food Industry(食品工业科技),2017,38(8):256-261.
[6]Satoshi H,Hideo E,Noriyuki Y,et al.Structure of resinosides from EucAlyptus resinifera as repellents against the blue mussel,Mytilus edulis[J].Bioscience,Biotechnology,and Biochemistry,1992,56(1):138.
[7]Pin D D,Chen Z T,Lee S W,et al.Antiproliferative activity and apopsisi-nduction of Eucalyptus citriodora resin and its major bioactive compound in melanoma B-16F10 cells[J].Journal of Agricultural and Food Chemistry,2012,60(32):7866-7872.
[8]Shen K H,Chen Z T,Duh P D.Cytotoxic effect of Eucalyptus citriodora resin on human hepatoma HepG2 cells[J].American Journal of Chinese Medicine,2012,40(2):399-413.
[9]Zhao Erlao(赵二劳),Yang Jie(杨洁),Zhao Sanhu(赵三虎).Research progress on extraction and purification technologies of flavonoids from peanut hull[J].Journal of The Chinese Cereals and Oils Association(中国粮油学报),2018,33(5):128-134.
[10]Rahul K L,Vaqar M A,Hasan M.Dietary flavonoid fisetin for cancer prevention and treatment[J].Molecular Nutrition&Food Research,2016,60(6):1401-1405.
[11]Du W J,Yang X L,Song Z J.Antitumor activity of total flavonoids from daphne genkwa in colorectal cancer[J].Phytotherapy Research,2015,30(2):323-330.
[12]Xu F,Wang C X,Wang H F,et al.Antimicrobial action of flavonoids from Sedum aizoon L.against lactic acid bacteria in vitro and in refrigerated fresh meat[J].Journal of Functional Foods,2018,40:744-750.
[13]Maria C Si,Laura N C,Franco M E,et al.Potent inhibition of western equine encephalitis virus by a fraction rich in flavonoids and phenolic acids obtained from achyrocline satureioides[J].Revista Brasileira de Farmacognosia,2016,26(5):571-578.
[14]de Souza P Oa,Bianch-Ib S E,Figueiróa F,et al.Anticancer activity of flavonoids isolated from achyrocline satureioides in gliomas cell lines[J].Toxicology in Vitro,2018,51:23-33.
[15]Zhao H,Song L,Huang W.Total flavonoids of epimedium reduce ageing related oxidative DNA damage in testis of rats via p53-depedent pathway[J].Andrologia,2017,49(10):1-7.
[16]Keti Z,Ketan R,Randolph R A,et al.Flavonoids and their metabolites:Prevention in cardiovascular diseases and diabetes[J].Diseases,2017,5(3):9.
[17]Jiang J,Xiao S C,Xua X M,et al.Isomeric flavonoid aglycones derivedfrom epimedii folium exerted different intensities in antiosteoporosis through OPG/RANKL protein targets[J].International Immunopharmacology,2018,62:277-286.
[18]Yi Y S.Regulatory roles of flavonoids on inflammasome activation during inflammatory responses[J].Molecular Nutrition&Food Research,2018,62(13):1-13.
[19]Yan T,Wu B,Liao Z Z,et al.Brainderived neurotrophic factor signaling mediates the a-ntidepressant like efect of the total favonoids of alpiniaeoxyphyllae fructus in chronic unpredictable mild stress mice[J].Phytotherapy Research,2016,30(9):1493-1502.
[20]Huang Liansha(黄链莎),Liu Tonghua(刘铜华),Sun Wen(孙文),et al.Effect and mechanism of flavonoids from mori folium on blood glucose level in diabetic[J].Chinese Journal of Experimental Traditional Medical Formulae(中国实验方剂学杂志),2018,24(16):152-156.
[21]Liang Q,Chen H G,Zhou X,et al.Optimized microwave-assistant extraction combined ultrasonic pretreatment of flavonoids from periplocaforrestiischltr.and evaluation of its antiallergic activity[J].Electrophoresis,2017,38(8):1113-1121.
[22]Lu Xiaoxiang(鲁晓翔).Research progress in antioxidant mechanism of flavonoids[J].Food Research and Development(食品研究与开发),2012,33(3):220-224.
[23]Yu Fuxiang(余付香),Chen Mingxu(陈明旭),Cheng Qiao(成桥),et al.Classified extraction and activity of total flavonoids from penthorum chinense purse[J].Nat Prod Res Dev(天然产物研究与开发).2017,29:976-982.
[24]Cai Jinyuan(蔡锦源),Wei Kunhua(韦坤华),Xiong Jianwen(熊建文),et al.Extraction of flavonoids from Sophora tonkinensis Gapnep and their antioxidant and antibacterial activities[J].Fine Chemicals(精细化工),2017,34(3):285-293.
[25]Wang Shasha(王沙沙),Liu Yang(刘洋),Niu Zhenzhen(牛真真),et al.Separation and purification of total flavonoids from salsolacollina with macroporous resins and evaluation of antioxidant activities[J].Fine Chemicals(精细化工),2018,35(1):58-64.
[26]Wang Fangfang(王芳芳).Studies on chemical constituents of dracaena cochinchinensis and their bioactivities[D].Kunming:Yunnan University of Nationalities(云南民族大学),2015.
[27]He Ying(何颖).The analysis of methods for extracting essential oils[J].Tianjin Pharmacy(天津药学),2015,27(1):47-50.
[28]Yao Xinsheng(姚新生).Natural pharmaceutical chemistry[M].Beijing:People’s Publishing House(人民出版社),1996:194.
[29]Ku Yongfeng(库咏峰),Huang Pinxian(黄品鲜),Chen Ping(陈萍),et al.Study on extrac-tion and separation analysis&antioxidant activity of total flavonoids from cinnamon[J].Application of Chemical(应用化工),2011,40(9):1547-1552.
[30]Zhou Yiming(周一鸣).Study on extraction purification and antioxidative effects of flavoniod sintatary bulk wheat bran[D].Xi'an:Shaanxi Normal University(陕西师范大学),2008.
[31]Zhou Ningbo(周宁波),Li Yujie(李玉杰),Yang Chengxiong(杨成雄),et al.Pharmaceutical analysis[M].Beijing:Chemical Industry Press(化学工业出版社),2010,7.
[32]Zeng Su(曾苏),Chai Yifeng(柴逸峰),Song Fenyun(宋粉云),et al.Pharmaceutical analysis[M].Beijing:Higher Education Press(高等教育出版社),2014:2.
[33]Xing Yun(邢云),Wang Guifang(王桂芳),Li Quanmin(李全民).Trends of solvent extraction in analytical chemistry[J].Physical and Chemical Testing-Chemical Section(理化检验-化学分册),2005,41(9):694-696.
[34]Wang P F,Mu X P,Du J J,et al.Flavonoid content and radical scavenging activity in fruits of Chinese dwarf cherry(C-erasus humilis)genotypes[J].Journal of Forestry Research,2018,29(1):55-63.
[35]Niveditha V R,Sridhar K R.Antioxidant activity of raw,cooked and Rhizopus oligosporus fermented beans of Canavalia of coastal sand dunes of Southwest India[J].Journal of Food Science and Technology,2014,51(11):3253-3260.
[36]Feng C Y,Su S,Wang L J,et al.Antioxidant capacities and anthocyanin ch-aracteristics of the black-red wild berries obt-ained in Northeast China[J].Food Chemistry,2016,204:150-158.
[37]Jeony S C,Jeony Y T,Yang B K,et al.White button mushroom(agaricusbisporus)lowers blood glucose and cholesterol levels in diabetic and hypercholesterolemic rats[J].Nutr Res,2010,30(1):49-56.
[38]Zhong Qiuping(钟秋平),Lin Meifang(林美芳).Determination on of contents of flavones and identifiction of flavones species in fructus clausenaelanii[J].Food Science(食品科学),2007,28(8):411-413.
[39]Feng Yabin(冯雅斌),Du Liang(杜靓),Wen Jing(温静).Study progress about drugs analysis on TLC[J].Disease Monitor&Control Jan(疾病监测与控制杂志),2011,5(1):60-63.
[40]Tian Qinglai(田庆来),Xie Yuchun(谢渝春),Zhang Bo(张波),et al.Separation of hydrophilic licorice flavonoid by solvent extraction method[J].The Chinese Journal of Process Engineering(过程工程学报),2007,7(3):496-500.
[41]Ni Xiaolei(倪晓蕾).Selective liquid-liquid extraction of natural phenolic compounds using ionic liquids[D].Hangzhou:Zhejiang University(浙江大学),2012.
[42]Marina P,Olga L T.Use of different spices as potential natural antioxidant additives on cooked beans(phaseol-us vulgaris).increase of DPPH radical scavenging activity and total phenolic content[J].Plant Foods for Human Nutrition,2014,69(4):337-343.
[43]Mariola S,Joanna N,Jerzy W,et al.Total antioxidant capacity of honey bee haemolymph in relation to age and exposure to pesticide,and comparison to antioxidant capacity of seminal plasma[J].Apidologie,2016,47(2):227-236.
[44]Narumol P,Weerasak S,Patcharawee N,et al.In vitro antioxidant activity study of novel chromone deriv-atives[J].Chemical Biology&Drug Design,2012,79(6):981-989.