基于LC-MS/MS法建立快速检测8种氨基糖苷类抗生素的方法

Rapid Detection of Eight Aminoglycoside Antibiotics by Liquid Chromatography-Tandem Mass Spectrometry

目的应用液质谱联用(LC-MS/MS)检索数据库技术建立卡那霉素、庆大霉素C1、庆大霉素C1a、庆大霉素C2、妥布霉素、新霉素、异帕米星、依替米星8种氨基糖苷类抗生素(aminoglycoside antibiotics,AGs)的快速检测方法。方法采用Shiseido PC HILIC(2.0 mm×100 mm,3μm)色谱柱对8种AGs进行分离,体积分数0.5%甲酸水溶液-0.1%甲酸乙腈溶液为流动相,梯度洗脱,流速为0.4 mL·min^(-1);采用多重反应监测(MRM)扫描建立8种AGs的筛查方法;采用碰撞能量扩展(CES)功能和增强子离子(EPI)扫描模式采集8种AGs的典型质谱图,添加到已经建立的抗生素检索质谱库中。结果所建立的方法专属性强,灵敏度高,10 min即可实现对8种AGs的初筛和定性分析。应用该方法对3批INSA胰岛素产品中的卡那霉素残留进行了监测,3批样品中均未检测到卡那霉素,其残留量小于检出限0.05 ng·mg^(-1)。结论本研究应用液质联用检索数据库技术建立的8种AGs的检测方法可以对痕量AGs进行快速筛查和准确定性。

OBJECTIVE To establish a method for simultaneous determination of 8 aminoglycoside antibiotics(AGs)including kanamycin,gentamycin C1,gentamycin C2,gentamycin C1a,tobramycin,neomycin,isepamicin and etimicin by LC-MS/MS with instant library searching.METHODS Eight AGs were separated by Shiseido PC HILIC(2.0 mm×100 mm,3μm)column,with 0.5%formic acid aqueous solution-0.1%formic acid acetonitrile solution as mobile phase and 0.4 mL·min~(-1)of flow rate,and procedure of the gradient elution was performed.The screening method of 8 AGs was established by the scanning mode of multiple reaction monitoring(MRM).Typical mass spectrograms of 8 AGs were collected by collision energy expansion(CES)function and enhanced ion(EPI)scanning mode,and added them to the already established mass spectralibrary of antibiotics searching.RESULTS The established method has the advantages of short analysis time,strong specificity and high sensitivity.The screening and qualitative analysis of 8 AGs can be realized within 10 min.Three batches of the INSA insulin product were detected by this method,and kanamycin that was less than the detection limit of 0.05 ng·mg~(-1)were not detected in above samples.CONCLUSION In this study,the detection method of 8 AGs established by LC-MS/MS with instant library searching can quickly screen and accurately determine trace AGs.