连翘果、连翘叶乙醇提取物的抑菌活性及成分分析
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摘要
为研究连翘果和连翘叶的抑菌活性及其活性成分的差异,采用滤纸片法测定连翘果、连翘叶乙醇提取物对三种细菌的抑菌圈直径,倍比稀释法测定了两种样品对金黄色葡萄球菌的最低抑菌浓度,并分析了两种样品对金黄色葡萄球菌的生长曲线及培养液中大分子物质和碱性磷酸酶的影响。采用气质联用分析了连翘果、连翘叶乙醇提取物中的挥发性成分。结果表明,连翘果与连翘叶乙醇提取物对大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌均有抑制效果,其中连翘叶乙醇提取物对三种细菌的抑菌圈直径分别为(12.7±1.2)、(15.0±0.8)、(14.7±0.7) mm,均大于连翘果乙醇提取物;连翘果、连翘叶乙醇提取物对金黄色葡萄球菌的最低抑菌浓度分别为6.25、3.125 mg/mL;连翘果、连翘叶乙醇提取物可以破坏细胞壁、细胞膜的完整性,使培养液中核酸等大分子物质以及碱性磷酸酶含量增加,并抑制金黄色葡萄球菌的生长繁殖,且连翘叶乙醇提取物的抑制作用大于连翘果乙醇提取物。气质联用分析发现连翘果乙醇提取物中主要包括醇类(38.55%)、酚类(14.00%)、酮类(11.07%)等物质;连翘叶乙醇提取物中主要包括醇类(34.97%)、烯烃类(12.71%)、酚类(12.26%)、烷烃类(11.05%)等;连翘果、连翘叶乙醇提取物中醇类、酚类、酮类、醛类、烷烃类物质对其抑菌活性有较大贡献,且其抑菌活性的差异主要与这些物质的种类和含量有关。
To study the antimicrobial activities and the difference of active constituents of Forsythia suspense fruit and Forsythia suspense leaves,the diameter inhibition zone of Forsythia suspensa fruit and Forsythia suspensa leaves ethanol extract against bacteria was measured by K-B method,the minimum inhibitory concentration (MIC) was measured by double dilution method.The antibacterial mechanism of Forsythia suspensa fruit and Forsythia suspensa leaves ethanol extract against Staphylococcus aureus was studied by the growth curves and the leakage of cell contents and alkaline phosphatase (AKP).Gas chromatography-mass spectrometry was used to analyze the chemical composition of Forsythia suspensa fruit and Forsythia suspensa leaves ethanol extract. The results showed that ethanol extract from Forsythia suspensa fruit and Forsythia suspensa leaves had antimicrobial effects on Escherichia coli,Staphylococcus aureus and Bacillus subtilis,the diameter inhibition zone of Forsythia suspensa leaves ethanol extract against these bacteria was (12.7 ± 1.2), (15.0 ± 0.8), (14.7 ± 0.7) mm,respectively.The MIC of Forsythia suspensa fruit and Forsythia suspensa leaves ethanol extract against Staphylococcus aureus was 6.25,3.125 mg/mL,respectively.The experimental results of antimicrobial mechanism showed that the ethanol extract from Forsythia suspensa fruit and Forsythia suspensa leaves inhibited the growth of Staphylococcus aureus,damaged the cell wall and cell membrane,increased the content of macromolecules and AKP in the culture solution,and the antimicrobial activity of Forsythia suspensa leaves ethanol extract was better than Forsythia suspensa fruit ethanol extract.GC-MS analysis showed that,ethanol extract from Forsythia suspensa fruit mainly included alcohols (38.55%),phenols (14.00%),and ketones (11.07%). Forsythia suspensa leaves ethanol extract contained alcohols (34.97%),olefins (12.71%),phenols (12.26%),and alkanes (11.05%). Alcohols,phenols,ketones,aldehydes and alkanes in the Forsythia suspensa fruit and Forsythia suspensa leaves ethanol extract had a significant contribution to their antibacterial activity,and the difference in their antibacterial activity was mainly related to the types and the content of these substances.
To study the antimicrobial activities and the difference of active constituents of Forsythia suspense fruit and Forsythia suspense leaves,the diameter inhibition zone of Forsythia suspensa fruit and Forsythia suspensa leaves ethanol extract against bacteria was measured by K-B method,the minimum inhibitory concentration (MIC) was measured by double dilution method.The antibacterial mechanism of Forsythia suspensa fruit and Forsythia suspensa leaves ethanol extract against Staphylococcus aureus was studied by the growth curves and the leakage of cell contents and alkaline phosphatase (AKP).Gas chromatography-mass spectrometry was used to analyze the chemical composition of Forsythia suspensa fruit and Forsythia suspensa leaves ethanol extract. The results showed that ethanol extract from Forsythia suspensa fruit and Forsythia suspensa leaves had antimicrobial effects on Escherichia coli,Staphylococcus aureus and Bacillus subtilis,the diameter inhibition zone of Forsythia suspensa leaves ethanol extract against these bacteria was (12.7 ± 1.2), (15.0 ± 0.8), (14.7 ± 0.7) mm,respectively.The MIC of Forsythia suspensa fruit and Forsythia suspensa leaves ethanol extract against Staphylococcus aureus was 6.25,3.125 mg/mL,respectively.The experimental results of antimicrobial mechanism showed that the ethanol extract from Forsythia suspensa fruit and Forsythia suspensa leaves inhibited the growth of Staphylococcus aureus,damaged the cell wall and cell membrane,increased the content of macromolecules and AKP in the culture solution,and the antimicrobial activity of Forsythia suspensa leaves ethanol extract was better than Forsythia suspensa fruit ethanol extract.GC-MS analysis showed that,ethanol extract from Forsythia suspensa fruit mainly included alcohols (38.55%),phenols (14.00%),and ketones (11.07%). Forsythia suspensa leaves ethanol extract contained alcohols (34.97%),olefins (12.71%),phenols (12.26%),and alkanes (11.05%). Alcohols,phenols,ketones,aldehydes and alkanes in the Forsythia suspensa fruit and Forsythia suspensa leaves ethanol extract had a significant contribution to their antibacterial activity,and the difference in their antibacterial activity was mainly related to the types and the content of these substances.
引文
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[3]房信胜,魏蕾,付欣欣,等.连翘长花柱与短花柱植株叶和果成分代谢研究[J].中药材,2018,41(3):567-572.
[4]梅雪,周安琴,李静,等.连翘叶的化学成分、药理学与毒理学研究概况[J].中国药房,2015,26(22):3143-3146.
[5]段飞,张双民,杨建雄,等.连翘叶提取物抑菌作用的研究[J].西北药学杂志,2005,20(2):66-67.
[6]秦臻,徐军,张立伟.连翘及连翘叶体外抑菌作用研究[J].食品工程,2013(2):49-52.
[7]原江锋,王大红,何灵美,等.连翘叶提取物抑菌作用以及稳定性研究[J].食品工业科技,2013,34(5):57-65.
[8]胡庭俊,温日清,赵灵颖.连翘不同极性段提取物制备及体外抑菌作用[J].江苏农业科学,2009(3):268-269.
[9]王浴生,邓文龙,薛春生.中药药理与应用[M].北京:人民卫生出版社,1983,516.
[10]葛康康,姚翰文,潘佳亮.山核桃干腐病拮抗细菌的鉴定及其抑菌效果[J].东北林业大学学报,2018,46(1):95-100.
[11]Filippini M,Ortelli C,Svercel M,et al.Interspecies variation in survival and growth of filamentous heterotrophic bacteria in response to UVC radiation[J].Journal of Photochemistry and Photobiology B:Biology,2011,103(3):234-242.
[12]Paul S,Dubey R C,Maheswari D K,et al.Trachyspermum ammi(L.)fruit essential oil influencing on membrane permeability and surface characteristics in inhibiting food-borne pathogens[J].Food Control,2011,22(5):725-731.
[13]张媛媛,冯亚净,李书国.五味子乙醇提取物的抑菌稳定性及抑菌机理研究[J].粮油食品科技,2016,24(1):55-60.
[14]孙京新,王文娟.多酚对假单胞菌抑菌机理研究[J].肉类研究,2009(10):48-51.
[15]Di Pasqua R,Betts G,Hoskins N,et al.Membrane toxicity o antimicrobial compounds from essential oils[J].Journal o agricultural and food chemistry,2007,55(12):4863-4870.
[16]张立华,王丹,宫文哲,等.枣核木醋液化学成分分析及其抑菌活性[J].食品科学,2016,37(14):123-127.
[17]秦娇.毛竹叶挥发油的提取及抑菌作用的研究[D].北京:北京林业大学,2010.