五味子对耐甲氧西林金黄色葡萄球菌体外抑菌活性部位的筛选和UP-LC-QTOF-MS/MS分析活性组分化学成分

Screening and UPLC-QTOF-MS/MS analysis of active fractions of Schisandra chinensis against methicillin-resistant Staphylococcus aureus in vitro

目的:研究五味子对耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA)体外抑菌活性部位的筛选以及分析五味子活性组分化学成分。方法:采用系统溶剂法分段萃取和AB-8大孔吸附树脂分离技术,结合管碟法对各分离组分进行体外抗耐甲氧西林金黄色葡萄球菌活性追踪检测,筛选出五味子抑菌活性最强组分。超高效液相(UPLC)、电喷雾四级杆飞行时间质谱(ESI-QTOF-MS)和二级质谱(MS/MS)对五味子抑菌活性组分进行分析。结果:筛选出70%乙醇提取的五味子依次经乙酸乙酯萃取和AB-8大孔吸附树脂水洗脱的组分(EEW component)为抗MRSA体外抑菌活性部位,同时对抗生素敏感金黄色葡萄球菌和临床分离的耐药MRSA均具有较强的抑菌活性。UPLC-QTOF-MS/MS分析体外抑菌活性组分,根据一级质谱提供的相对分子质量和二级质谱高能量碰撞碎片信息,初步鉴定了11个化合物,这11个化合物分别为奎尼酸(1)、苹果酸(2)、柠檬酸(3)、苹果酸异构体(4)、咖啡酰奎尼酸(5)、莽草酸(6)、柠檬酸单甲酯(7)、原儿茶酸(8)、富马酸单乙酯(9)、2-羟基-丙基三羧酸-2-乙酯(10)和柠檬酸二甲酯(11)。结论:本实验初步筛选了五味子抑菌活性部位,为开发天然抑菌制剂提供了方法和思路。

OBJECTIVE To screen the antibacterial active parts of Schisandra chinensis against methicillin-resistant Staphylococcus aureus and to analyze the chemical constituents of its active components.METHODS By systematic solvent extraction and separation technology of AB-8 macroporous adsorption resin combined with cylindekplate method for the separation of components against methicillin-resistant Staphylococcus aureus detection,the strongest antibacterial activity components of Schisandra chinensis were screen out.UPLC,ESI-QTOF-MS and MS/MS were used to analyze the antibacterial components of Schisandra chinensis.RESULTS 70% ethanol extracts of Schisandra chinensis extracted by ethyl acetate and AB-8 macroporous resin water elution fractions（EEW Component）were screened out as effective sites against MRSAin vitro,while MRSA was resistant to antibiotic sensitive Staphylococcus aureus and clinical isolates had strong antibacterial activity.Meanwhile,the active fractions also showed strong antibacterial activity against antibiotic sensitive Staphylococcus aureus and the clinically isolated drug-resistant MRSA.According to the molecular weight and high energy collision debris information obtained by UPLC-QTOF-MS/MS,the active components were characterized by 11 large polar compounds.The organic acids were identified as quinic acid（1）,malic acid（2）,citric acid（3）,malic acid isomer（4）,caffeoyl guinic acid（5）,shikimic acid（6）,2-methyl citrate（7）,protocatechuic acid（8）and fumaric acid monoethyl ester（9）,2-hydroxy-propane tricarboxylic acid-2-ethyl ester（10）,1,5-dimethyl citrate（11）.CONCLUSION This experiment has preliminarily screened the antibacterial active parts of Schisandra chinensis and provided a method and idea for the development of natural antibacterial agents.