超高效液相色谱-串联质谱法测定水源水中6种β-内酰胺类抗生素的方法优化
Optimization of six β-lactam antibiotics determination in source water by UHPLC-MS/MS
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摘要:
背景 目前对水体中β-内酰胺类抗生素的检测并未形成统一的标准方法，常用的检测方法污染重、危害大、耗时久。针对公共卫生突发事件，需要快速有效的检测方法。
目的 建立利用超高效液相色谱串联质谱法快速测定水源水中6种β-内酰胺类抗生素药物（阿莫西林、头孢他啶、头孢克洛、头孢拉定、青霉素V钾和氯唑西林）残留的检测方法。
方法 利用相对响应值评价基质效应。比较不同温度和保存时间水源水中β-内酰胺类药物的稳定性，确定最佳保存条件。水样经0.22μm微孔滤膜过滤，采用大体积进样方式，以0.1%甲酸水溶液和0.1%甲酸乙腈溶液作为流动相进行梯度洗脱，在Kinetex 2.6 μm C18色谱柱上进行分离，采用电喷雾离子源正离子和多反应监测模式进行检测。
结果 通过基质曲线补偿基质效应的干扰。水样于采样瓶内4℃保存，24 h内进行分析可避免目标物降解。6种β-内酰胺类抗生素在线性范围内线性关系良好，检出限（信噪比S/N=3）为4~7 ng/L，定量限（S/N=10）为10~20 ng/L，加标回收率为84.8%~99.6%，相对标准偏差小于等于15.6%（n=6）。
结论 该方法简化了前处理过程，可以满足应急事件中同时检测水源水中6种β-内酰胺类抗生素残留的要求。
Abstract:Background At present, no generally accepted standard methods are available for detection of β-lactam antibiotics in water, while conventional methods are polluting, harmful, and timeconsuming. Rapid and effective detection methods are therefore warranted in handling public health emergencies.
Objective This study aims to establish a rapid method for detecting six β-lactam antibiotic residues in source water by ultra high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS), including amoxicillin, ceftazidime, cefaclor, cephradine, phenoxymethylpenicillin potassium, and cloxacillin.
Methods Matrix effects were evaluated with relative response value. The best storage conditions were determined by comparing the stability of β-lactam drugs in source water at different temperatures combined with different storage time. Water samples were filtered through a 0.22 μm rated microporous membrane using large volume injection, and then separated on Kinetex 2.6 μm C18 column with gradient elution using mobile phase of 0.1% formic acid and 0.1% acetonitrile. Electrospray ionization in positive mode and multiple reaction monitoring mode were used for detection.
Results Matrix effect interference was compensated by matrix curve. The water samples were stored at 4℃ and analyzed within 24 h to avoid degradation of target chemicals. The six β-lactam antibiotics showed good linear relationships in designed ranges. The limits of detection (S/N=3) were 4-7 ng/L, the limits of quantification (S/N=10) were 10-20ng/L, the adding standard recoveries were 84.8%-99.6%, and the relative standard deviations were all less than 15.6% or equal to (n=6).
Conclusion The method is featured with simplified pre-treatment process and meets the requirement of simultaneously detecting six β-lactam antibiotic residues in source water in case of emergency events.
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