高效液相色谱-串联质谱法同时测定细菌群体感应效应的11种AHLs类信号分子  

Detection of N-Acyl-homoserine Lactones Class Signal Molecules of Quorum Sensing Secreted by Bacteria Using High Performance Liquid Chromatography-Mass Spectrometry/Mass Spectrometry

作　　者：马晨晨[1] 李柏林[1] 欧杰[1] 王婧[1] 赵俊虹[2] 

机构地区：[1]上海海洋大学食品学院,上海201306 [2]上海市食品研究所,上海200235

出　　处：《分析化学》2010年第10期1428-1432,共5页Chinese Journal of Analytical Chemistry

基　　金：国家"十一五"科技支撑计划项目(No.2007BAD52B05);上海市科委应用技术开发专项基金项目(No.09-223)资助

摘　　要：建立了高效液相色谱-串联质谱(HPLC-MS/MS)法同时测定细菌群体感应分泌的11种AHLs类信号分子的分析方法。细菌所分泌AHLs类信号分子经乙酸乙酯萃取后简单处理,可直接进样分析。采用Sunfire C18色谱柱(50mm×2.1mm,3.5μm),甲醇与水(含2mmoI/L乙酸铵,0.1%甲酸)梯度进样,采用电喷雾离子源正离子模式,多反应监测(MRM)模式,外标法进行质谱定量分析。分析时间为12min。本方法在1～250μg/L范围内呈良好的线性关系(r>0.996);检出限为0.1～1.0μg/kg;定量限为0.3～3.0μg/kg;平均添加回收率为54.4%～128.6%;相对标准偏差为3.5%～13.9%。本方法具有操作简便、快速、准确、灵敏度高等优点,适用于多种AHLs类信号分子化合物的同时定性与定量分析,为微生物群体感应的进一步研究提供了有效的且便利的分析方法。A confirmative method was developed with high performance liquid chromatography-mass spectrometry（HPLC-MS/MS） to detect acyl-homoserine lactoned（AHLs） secreted by bacterial quorum sensing.The AHLs secreted by bacterial quorum sensing were extracted with ethyl acetate.The samples could be directly analyzed after a simple treatment.Sunfire C18column（2.1 mm × 50 mm,3.5 μm） with methanol and water （including 2 mmoI/L ammonium acetate,0.1% formic acid） as the mobile phase was used for separation.The detection method of electrospray ionization source with positive ion was used for mass spectrometric analysis.Qualitative and quantitative analysis of 11 kinds of AHLs were completed within 12 min.Experimental results showed that,the AHLs was in good linearity（r 0.996） in the range of 1-250 μg/L.The detection limit of the method was 0.1-1.0 μg/kg.The limits of quantification were 0.3-3.0 μg/kg.The average recoveries were 54.4%-128.6% .The relative standard deviations were 3.5%-13.9% .The results show that,this method is simple,rapid,accurate,and highly sensitive to analyse and detect the AHLs in the bacterial growth environment.

关 键 词：高效液相色谱-串联质谱 细菌 群体感应效应 类信号分子 N-酰基高丝氨酸内酯 

分 类 号：Q93-3[生物学—微生物学]