微塑料生物膜11种AHLs类群体感应信号分子测定及其分泌特征  

Determination and secretion characteristics of 11 N-acyl-homoserine lactones signal molecules of quorum sensing in microplastic biofilms

作　　者：喻红霞 刘晓薇 汪慧香 王静 俞志敏[1,2] 邓呈逊 YU Hongxia;LIU Xiaowei;WANG Huixiang;WANG Jing;YU Zhimin;DENG Chengxun(School of Biology,Food,and Environment,Hefei University,Hefei,230601,China;International(Sino-German)Joint Research Center for Biomass of Anhui Province,Hefei,230601,China;School of Resources and Environmental Engineering,Hefei University of Technology,Hefei,230009,China)

机构地区：[1]合肥学院生物食品与环境学院,合肥230601 [2]安徽省生物质(中德)国际联合研究中心,合肥230601 [3]合肥工业大学资源与环境工程学院,合肥230009

出　　处：《环境化学》2022年第3期785-792,共8页Environmental Chemistry

基　　金：国家自然科学基金(52070063);巢湖生物资源调查及生态修复示范工程(2021BFFBZ01571)资助。

摘　　要：为研究微塑料生物膜的群体感应效应,利用液液萃取和超高效液相色谱-串联质谱技术,建立了微塑料上枯草芽孢杆菌生物膜分泌的11种酰基高丝氨酸内酯(AHLs)类群体感应信号分子前处理和仪器分析方法。优化后的前处理方法采用酸化的乙酸乙酯(0.01%冰乙酸,V/V)对微塑料生物膜样品中AHLs类信号分子进行萃取,萃取方式采用冰浴超声。微塑料生物膜中AHLs回收率为50.6%—128.1%,相对标准偏差(n=3)为0.2%—10.6%。对色谱和质谱条件进行优化,11种AHLs在0.5—50μg·L^(−1)范围内呈良好的线性关系(R2>0.995),检出限为0.005—0.01μg·L^(−1),定量限为0.01—0.02μg·L^(−1)。本方法具有快速、准确、灵敏度高等优点。对聚丙烯、聚乙烯、聚苯乙烯等3种微塑料上枯草芽孢杆菌生物膜中11种AHLs分析结果表明,C8-HSL、C12-HSL和3-oxo-C10-HSL在3种微塑料生物膜中均被检出;C12-HSL的产生量最高,且在聚乙烯表面生物膜中含量最高达34 ng·g^(−1)。通过添加外源信号分子研究表明,AHLs分子浓度增加会促进微塑料生物膜形成。In order to study the quorum sensing effect of biofilms on microplastics,a pretreatment and instrumental analysis method for 11 N-acyl-homoserine lactones(AHLs)secreted by Bacillus subtilis biofilms on microplastics was established by liquid-liquid extraction and ultra-performance liquid chromatography-tandem mass spectrometry.The optimized pretreatment method used acidified ethyl acetate(0.01%glacial acetic acid,V/V)to extract AHLs signal molecules from microplastic biofilm samples.The extraction method used ultrasound with ice bath.The recoveries of 11 AHLs in microplastic biofilms ranged from 50.6%to 128.1%,and the relative standard deviations(n=3)ranged from 0.2%to 10.6%.After the optimization for chromatographic and mass spectrometry condition,the linearities of 11 AHLs were good in the range of 0.5—50μg·L−1(R2>0.995),the limits of detection were in the range of 0.005—0.01μg·L−1,and the limits of quantification were in the range of 0.01—0.02μg·L−1.The method is rapid and accurate,and has high sensitivity.The quantitative analysis for 11 AHLs in Bacillus subtilis biofilms on polypropylene,polyethylene and polystyrene showed that C8-HSL,C12-HSL and 3-oxo-C10-HSL were all detected in the three types of microplastic biofilms;the production amount of C12-HSL was the highest,and was up to 34 ng·g−1 in polyethylene biofilm.The result from the addition of exogenous AHLs indicated the increasing concentrations of AHLs can promote the biofilm formation on microplastics.

关 键 词：超高效液相色谱-串联质谱 生物膜 群体感应 酰基高丝氨酸内酯 微塑料 

分 类 号：X505[环境科学与工程—环境工程]