地肤子抑菌活性成分筛选
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摘要
以最小抑菌浓度为指标,采用二倍稀释法分别测试地肤子乙醇提取物中的石油醚相、乙酸乙酯相、正丁醇相和水相萃取物对4株细菌和5株植物病原菌的抑制作用;再对抑菌活性较强的萃取物进行分离纯化,通过二倍稀释法测试所分离化合物的抑菌活性,并对其结构进行表征。结果表明,正丁醇相萃取物的抑菌活性最强,从中分离得到3个化合物,其中化合物Ⅰ的抑菌活性较强,对大肠杆菌具有显著的抑制作用,其最小抑菌浓度为31.3μg·mL-1,经1 HNMR、IR及高分辨质谱等鉴定化合物Ⅰ为齐墩果酸。齐墩果酸有较强的抑菌活性,是地肤子抑菌活性的物质基础。
Using the minimal antibacterial concentration as an index,we determined the antibacterial activities of four fractions from ethanol extract against four bacteria and five plant pathogenous fungi by a double-dilution method,such as petroleum ether fraction,acetic ether fraction,n-butanol fraction,and water fraction.Moreover,we separated and purified a fraction with higher antibacterial activity,determined the antibacterial activities of purified compounds by a double-dilution method,and characterized the structures of the compounds.The results indicate that n-butanol fraction shows the highest antibacterial activity,from which three compounds are separated.Among them,compoundⅠhas the highest antibacterial activity,which shows strong antibacterial activity against Escherichia coli with the minimal antibacterial concentration of 31.3μg·mL~(-1).Furthermore,compoundⅠis identified as oleanic acid by ~1HNMR,IR,and high resolution mass spectrometer.Oleanic acid has high antibacterial activity,which is the material basis of the antibacterial activity of Fructus kochiae.
Using the minimal antibacterial concentration as an index,we determined the antibacterial activities of four fractions from ethanol extract against four bacteria and five plant pathogenous fungi by a double-dilution method,such as petroleum ether fraction,acetic ether fraction,n-butanol fraction,and water fraction.Moreover,we separated and purified a fraction with higher antibacterial activity,determined the antibacterial activities of purified compounds by a double-dilution method,and characterized the structures of the compounds.The results indicate that n-butanol fraction shows the highest antibacterial activity,from which three compounds are separated.Among them,compoundⅠhas the highest antibacterial activity,which shows strong antibacterial activity against Escherichia coli with the minimal antibacterial concentration of 31.3μg·mL~(-1).Furthermore,compoundⅠis identified as oleanic acid by ~1HNMR,IR,and high resolution mass spectrometer.Oleanic acid has high antibacterial activity,which is the material basis of the antibacterial activity of Fructus kochiae.
引文
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[2]吕选民,姬水英.柴草瓜果篇第十七讲地肤子[J].中国乡村医药,2016,23(21):48-49.
[3]陈雪羽.地肤子的化学成分·药理学研究进展[J].安徽农业科学,2010,38(21):11138-11139.CHEN X Y.Progress of Fructus kuchiae in fields of chemical components and pharmacological study[J].Journal of Anhui Agricultural Sciences,2010,38(21):11138-11139.
[4]中国科学院《中国植物志》编辑委员会.中国植物志[M].北京:科学出版社,1979:102.
[5]林秀仙,李菁.超临界萃取地肤子油的抑菌作用研究[J].中药材,2004,27(8):603-604.
[6]吴静,师光禄,苏学友,等.地肤子提取物对几种植物病原菌生物活性的影响[J].北京农学院学报,2008,23(3):36-39.WU J,SHI G L,SU X Y,et al.A preliminary study on bioactivity of extracts from Fructus kochiae against several phytopathogens[J].Journal of Beijing Agricultural College,2008,23(3):36-39.
[7]赵金凤,胡功政,苑丽,等.菌悬液浓度对体外药敏实验结果的影响[J].实验科学与技术,2012,10(4):33-35.ZHAO J F,HU G Z,YUAN L,et al.Impact of bacteria suspensions on the antibiotic susceptibility test results[J].Experiment Science&Technology,2012,10(4):33-35.
[8] BHARATE S B,KHAN S I,YUNUS N M,et al.Antiprotozoal and antimicrobial activities of O-alkylated and formylated acylphloroglucinols[J].Bioorganic and Medicinal Chemistry,2007,15(1):87-96.
[9]张起辉,高慧媛,吴立军,等.吴茱萸的化学成分[J].沈阳药科大学学报,2005,22(1):12-14.ZHANG Q H,GAO H Y,WU L J,et al.Chemical constituents of Evodiarutaecarpa(Juss.)Benth.[J].Journal of Shenyang Pharmaceutical University,2005,22(1):12-14.
[10]薛芊,郭美丽,张戈.青箱子化学成分研究[J].药学服务与研究,2006,6(5):346-347.XUE Q,GUO M L,ZHANG G.Study of chemical constituents of Semen celosiae[J].Pharmaceutical Care and Research,2006,6(5):346-347.
[11]肖怀,刘光明,王卓,等.丽江獐牙菜(Swettia delavayi)化学成分研究[J].大理学院学报,2006,5(6):17-19.XIAO H,LIU G M,WANG Z,et al.Studies on the chemical constituents of Swettia delavayi[J].Journal of Dali University,2006,5(6):17-19.
[12]柏玉冰,李春,周亚敏,等.夏枯草的化学成分及其三萜成分的抗肿瘤活性研究[J].中草药,2015,46(24):3623-3629.BAI Y B,LI C,ZHOU Y M,et al.Chemical constituents of triterpenoids from Prunella vulgaris and their antitumor activities[J].Chinese Traditional and Herbal Drugs,2015,46(24):3623-3629.