薄层色谱-生物自显影观察新疆两色金鸡菊中清除自由基和抑制α-葡萄糖苷酶活性成分
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摘要
目的:采用薄层色谱-生物自显影技术(TLC-bioautography),观察新疆两色金鸡菊提取物中具有清除自由基和抑制α-葡萄糖苷酶活性成分.方法:以1,1'-二苯基苦基苯肼(DPPH)和2,2'-氨基-二(3-乙基-苯并噻唑啉-6-磺酸)二铵盐(ABTS)为显色剂,观察新疆两色金鸡菊水提物和醇提物中清除自由基的有效成分;以对硝基苯-β-D-葡萄糖苷(p-Nitrophanyl-β-D-Gluopyranoside,p NPG)为底物,观察水提物和醇提物中具有抑制α-葡萄糖苷酶(来源于啤酒酵母菌)的活性成分.采用混合标准物质TLC色谱进行活性成分定性分析.结果:新疆两色金鸡菊水提物和醇提物中均含有马里苷、奥卡宁、栎草亭-7-O-β-D-葡萄糖苷、黄酮奥卡宁(flavanokanin)、绿原酸、芦丁和槲皮素等化学成分.经薄层色谱-生物自显影实验,两色金鸡菊水提物和醇提物中的黄酮奥卡宁、马里苷、芦丁和绿原酸均有较强清除DPPH和ABTS自由基的能力;水提物和醇提物中黄酮奥卡宁、奥卡宁和马里苷能有效抑制α-葡萄糖苷酶的活性.结论:经薄层色谱-生物自显影技术可快速发现两色金鸡菊中具有清除自由基和抑制α-葡萄糖苷酶活性成分,为进一步研究提供了指引.
Aim: To study the active components of Coreopsis tinctoria extract by TLC-bioautography for their free radical scavenging capability and α-glucosidase inhibitory activity. Method: the free radical scavenging activity of active components of C. tinctoria Nutt. was determined by TLC-bioautography,and the color reagent used as 1,1'-diphenyl-2-picrylhydrazyl(DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid diammonium salt,ABTS),respectively. In α-glucosidase inhibitory assay,theα-glucosidase(from Saccharomyces cerevisiae) and the p-Nitrophanyl-β-D-gluopyranoside(p NPG) were used as the enzyme and substrate. The qualitative analysis of the active components was performed by TLC chromatography with mixed standard substances as references. Results: the seven chemical constituents were identified in C. tinctoria aqueous extract and alcoholic extract such as marein,okanin,quercetagetin-7-O-β-D-glucopyranoside, flavanokanin, chlorogenic acid, rutin and quercetin. Theflavanokanin,marein,rutin and chlorogenic acid in aqueous extract and alcoholic extract showed stronger ability to scavenge DPPH and ABTS free radicals using TLC-bioautography analysis. The active constituents of α-glucosidase inhibitors were flavanokanin,okanin and marein in aqueous extract and alcoholic extract. Conclusion: the active components with free radical scavenging capability and α-glucosidase inhibitory activity can quickly find by TLC-bioautography,and this technology provides guidance for further study of C. tinctoria.
Aim: To study the active components of Coreopsis tinctoria extract by TLC-bioautography for their free radical scavenging capability and α-glucosidase inhibitory activity. Method: the free radical scavenging activity of active components of C. tinctoria Nutt. was determined by TLC-bioautography,and the color reagent used as 1,1'-diphenyl-2-picrylhydrazyl(DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid diammonium salt,ABTS),respectively. In α-glucosidase inhibitory assay,theα-glucosidase(from Saccharomyces cerevisiae) and the p-Nitrophanyl-β-D-gluopyranoside(p NPG) were used as the enzyme and substrate. The qualitative analysis of the active components was performed by TLC chromatography with mixed standard substances as references. Results: the seven chemical constituents were identified in C. tinctoria aqueous extract and alcoholic extract such as marein,okanin,quercetagetin-7-O-β-D-glucopyranoside, flavanokanin, chlorogenic acid, rutin and quercetin. Theflavanokanin,marein,rutin and chlorogenic acid in aqueous extract and alcoholic extract showed stronger ability to scavenge DPPH and ABTS free radicals using TLC-bioautography analysis. The active constituents of α-glucosidase inhibitors were flavanokanin,okanin and marein in aqueous extract and alcoholic extract. Conclusion: the active components with free radical scavenging capability and α-glucosidase inhibitory activity can quickly find by TLC-bioautography,and this technology provides guidance for further study of C. tinctoria.
引文
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[2]谷丽华,侴桂新,王峥涛.基于ABTS自由基的薄层色谱-生物自显影新方法的建立及其在中药抗氧化活性评价中的应用[J].上海中医药大学学报,2013,27(3):99-104.GU L H,CHOU G X,WANG Z T.Establishment and application of a new TLC-bioautography technology in antioxidant activity evaluation of materia medica[J].Acta Universitatis Traditionis Medicalis Sinensis Pharmacologiaeque Shanghai,2013,27(3):99-104.
[3]王双英,穆青.薄层色谱直接生物自显影方法(TLC-bioautography)在抗菌剂分析中的应用[J].国外医药(抗生素分册),2012,33(2):67-69.WANG S Y,MU Q.Application of TLC-bioautography in Analysis of antibacterial agents[J].Foreign Medicine(Antibiotics volume),2012,33(2):67-69.
[4]韩松林,李新霞,勉强辉,等.薄层生物自显影技术比较新疆2种洋甘菊抗氧化活性[J].中国中药杂志,2013,38(2):193-198.HAN S L,Li X X,MIAN Q H,et al.Comparison of antioxidant activity between two species of chamomiles produced in Xinjiang by TLC-bioautography[J].China journal of Chinese Material Medica,2013,38(2):193-198.
[5]新疆植物志编辑委员会.新疆植物志(第5卷)[M].乌鲁木齐:新疆科技卫生出版社,1999:93.Commissione Redactorum Flora Xinjiangensis(Tomus 5)[M].Urumqi:Xinjiang Science&Technology&Hygiene Publishing House,1999:93.
[6]张文广,李琳琳,王烨,等.新疆昆仑雪菊提取物对糖尿病小鼠血糖的影响[J].中国药物应用与监测,2015,12(2):82-84.ZHANG W G,LI L L,WANG Y,et al.Effects of extracts from Xinjiang Kunlun Coreopsis tinctoria flowers on blood glucose in diabetic mice[J].Chinese Journal of Drug Application and Monitoring,2015,12(2):82-84.
[7]毛新民,卢伟,李琳琳,等.两色金鸡菊化学成分和药理作用研究进展[J].中国药物应用与监测,2014,11(4):235-239.MAO X M,LU W,LI L L,et al.The progress of chemical constituents and pharmacological action of Coreopsis tinctoria Nutt[J].Chinese Journal of Drug Application and Monitoring,2014,11(4):235-239.
[8]梁乐,李琳琳,古扎力努尔·艾尔肯,等.薄层生物自显影技术筛选两色金鸡菊中的活性成分[J].药物分析杂志,2015,35(3):425-429.LIANG L,LI L L,Guzhalinuer A,et al.TLCbioautographic method for screening of active constituents of Coreopsis tinctoria[J].Chinese Journal of Pharmaceutical Analysis,2015,35(3):425-429.
[9]姚新成,王新兵,张婷,等.新疆两色金鸡菊总黄酮含量测定显色反应体系.暨南大学学报(自然科学与医学版),2015,36(3):222-227.YAO X C,WANG X B,ZHANG T,et al.The determination method of total flavonoids in Coreopsis tinctoria Nutt.from Xinjiang in different chromogenic reaction system by spectrophotometry[J].Journal of Jinan University(Natural Science&Medicine Edition),2015,36(3):222-227.