摘要
为了开发一种高效液相色谱-质谱联用(HPLC-MS)方法,用于定量检测细菌内源性低浓度的N-酰基高丝氨酸内酯(AHLs),本研究通过液液萃取法提取细菌内源产生的AHLs,并优化了质谱和色谱条件,建立了一套具有良好线性关系、较小基质效应和较高回收率的检测体系。结果表明,7种AHLs的线性相关系数(r)均大于0.995,所有AHLs的r均大于0.990,表明该方法具有良好的线性关系。8种AHLs的基质效应均大于80.0%,细菌生长基质对内源AHLs的检测影响较小。研究还发现碳链较长的AHLs在复杂基质中的干扰较小,而含羰基的AHLs表现出一定的信号抑制效应。7种AHLs的回收率均超过80.0%,显示了该方法具有高回收率和良好的准确性及可靠性。通过该方法检测细菌内源性AHLs,结果显示C_(10)-HSL和C_(12)-HSL是主要的AHLs,进一步验证了方法的有效性。此方法能够满足对细菌内源微量AHLs的精确检测,为研究细菌群体感应(QS)系统中的内源AHLs提供了可靠的技术支持。
In order to develop a high-performance liquid chromatography-mass spectrometry(HPLC-MS)method for quantitatively detecting endogenous low concentrations of N-acyl homoserine lactones(AHLs)in bacteria,this study extracted AHLs produced by bacteria through liquid-liquid extraction,optimized mass spectrometry and chromatographic conditions,and established a detection system with good linear relationship,small matrix effect,and high recovery rate.The results showed that the linear correlation coefficients(r)of the seven AHLs were all greater than 0.995,and the r values of all AHLs were greater than 0.990,indicating that the method has a good linear relationship.The substrate effects of 8 types of AHLs are all greater than 80.0%,and the bacterial growth matrix has a relatively small impact on the detection of endogenous AHLs.The study also found that AHLs with longer carbon chains have less interference in complex matrices,while AHLs containing carbonyl groups exhibit a certain signal suppression effect.The recovery rates of 7 types of AHLs all exceeded 80.0%,indicating that this method has high recovery rates and good accuracy and reliability.The method was used to detect endogenous AHLs in bacteria,and the results showed that C_(10)-HSL and C_(12)-HSL were the main AHLs,further verifying the effectiveness of the method.This method can accurately detect endogenous trace AHLs in bacteria,providing reliable technical support for studying endogenous AHLs in bacterial quorum sensing(QS)systems.
作者
赵文洁
陈昢圳
张艳伟
李少朋
ZHAO Wenjie;CHEN Peizhen;ZHANG Yanwei;LI Shaopeng(Agro-Environmental Protection Institute,Ministry of Agriculture and Rural Affairs,Tianjin 300191,China;Tianjin Agricultural University,Tianjin,300384,China)
出处
《农业环境科学学报》
北大核心
2025年第9期2478-2488,共11页
Journal of Agro-Environment Science
基金
天津市科技计划项目(22YFZCSN00170)
中国农业科学院创新工程重大科研任务(CAAS-ZDRW202306)。
作者简介
赵文洁(1997-),女,山西晋城人,硕士研究生,研究方向为农村生活污水生物强化治理。E-mail:ZWJ1997JC@163.com;通信作者:陈昢圳,E-mail:chenpeizhen2024@163.com;通信作者:李少朋,E-mail:lishaopeng518896@163.com。
