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
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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 2.6μm 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。
[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 10-20 ng/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.
[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 10-20 ng/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.
引文
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[8]彭凡立,吉文亮,彭丹红,等.超高效液相色谱串联质谱检测尿液中6种环境内分泌干扰物方法的建立与应用[J].环境与职业医学,2016,33(5):499-503.
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[11]张秀尧,蔡欣欣.超高效液相色谱-三重四极杆质谱法快速同时测定牛奶中53种β-内酰胺类抗生素及其代谢产物的残留[J].色谱,2014,32(7):693-701.
[12]刘洪斌,文一,刘勇军,等.LC-MS/MS法检测牛奶中14种β-内酰胺类抗生素残留[J].分析试验室,2014,33(2):221-224.
[13]生活饮用水标准检验方法水样的采集和保存:GB/T5750.2-2006[S].北京:中国标准出版社,2007.
[14]MATUSZEWSKI B K,CONSTANZER M L,CHAVEZ-ENG C M.Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS[J].Anal Chem,2003,75(13):3019-3030.
[15]KACZY?SKI P.Clean-up and matrix effect in LC-MS/MSanalysis of food of plant origin for high polar herbicides[J].Food Chem,2017,230:524-531.
[2]龚勇青.水中抗生素检测意义及检测技术综述[J].科技资讯,2017,15(28):253-254.
[3]生活饮用水卫生标准:GB 5749-2006[S].北京:中国标准出版社,2007.
[4]叶能胜,谷学新,张琦.固相萃取-高效液相色谱法检测牛奶和猪肉中5种β-内酰胺类抗生素[J].分析实验室,2010,29(6):73-77.
[5]李勇竞,郑妹凤.固相萃取-超高效液相色谱法测定饮用水中四环素类抗生素[J].环境与职业医学,2016,33(6):624-626.
[6]朱峰,吉文亮,阮丽萍,等.高效液相色谱-质谱联用法同时检测水体中13种β-内酰胺类药物残留[J].色谱,2016,34(3):299-305.
[7]陆克祥,隋铭皓,高乃云.固相萃取-超高压液相色谱-串联质谱测定水中19种抗生素[J].分析测试学报,2010,29(12):1209-1214.
[8]彭凡立,吉文亮,彭丹红,等.超高效液相色谱串联质谱检测尿液中6种环境内分泌干扰物方法的建立与应用[J].环境与职业医学,2016,33(5):499-503.
[9]HERNáNDEZ F,CALISTO-ULLOA N,GóMEZ-FUENTES C,et al.Occurrence of antibiotics and bacterial resistance in wastewater and sea water from the Antarctic[J].J Hazard Mater,2019,363:447-456.
[10]岳秀英,彭莉,程江,等.酶联免疫吸附法测定牛奶中青霉素残留的试验报告[J].四川畜牧兽医,2007,34(6):27-28.
[11]张秀尧,蔡欣欣.超高效液相色谱-三重四极杆质谱法快速同时测定牛奶中53种β-内酰胺类抗生素及其代谢产物的残留[J].色谱,2014,32(7):693-701.
[12]刘洪斌,文一,刘勇军,等.LC-MS/MS法检测牛奶中14种β-内酰胺类抗生素残留[J].分析试验室,2014,33(2):221-224.
[13]生活饮用水标准检验方法水样的采集和保存:GB/T5750.2-2006[S].北京:中国标准出版社,2007.
[14]MATUSZEWSKI B K,CONSTANZER M L,CHAVEZ-ENG C M.Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS[J].Anal Chem,2003,75(13):3019-3030.
[15]KACZY?SKI P.Clean-up and matrix effect in LC-MS/MSanalysis of food of plant origin for high polar herbicides[J].Food Chem,2017,230:524-531.