超高效液相色谱-串联质谱法检测鱼肉中48种抗生素

Determination of 48 kinds of antibiotics in fish muscle by ultra performance liquid chromatography-tandem mass spectrometry

目的建立超声波提取、固相萃取柱净化,结合超高效液相色谱-串联质谱法检测鱼肉中大环内酯类、喹诺酮类、磺胺类、四环素类和酰胺醇类等5大类48种抗生素的检测方法。方法样品分别经乙二胺四乙酸二钠盐-磷酸盐缓冲液(ethylene diamine tetraacetic acid disodium salt-Mcllvaine,Na_(2)EDTA-Mcllvaine)、磷酸盐缓冲液、乙腈/磷酸盐缓冲液进行提取,合并提取液稀释至100mL,经亲水亲酯平衡柱(hydrophilic-lipophilic balance,HLB)固相萃取净化后,氮吹至近干后定容待测。以0.1%甲酸甲醇和0.1%甲酸溶液为流动相,采用BEH C_(18)超高效液相色谱柱分离,采用多反应监测正负离子切换模式检测。结果该方法测定48种抗生素线性关系良好,决定系数均大于等于0.9922,方法检出限为0.02~0.47μg/kg,定量限为0.07~1.56μg/kg。3个加标水平的回收率在69.2%~120.3%之间,相对标准偏差范围为1.0%~14.3%。实际样品的检测结果也与标准方法的结果一致。结论该方法简便快速、灵敏可靠,适用于鱼肉中5大类48种抗生素药物残留的同时测定。

Objective To develop a method for detecting 48 kinds of antibiotics across 5 categories(macrolides,quinolones,sulfonamides,tetracyclines,and amide antibiotics)in fish meat using ultrasonic extraction,solid-phase extraction purification,and ultra performance liquid chromatography-tandem mass spectrometry.Methods Samples were extracted using ethylene diamine tetraacetic acid disodium salt-Mcllvaine(Na_2EDTA-Mcllvaine)buffer solution,phosphate buffer,and a phosphate buffer-acetonitrile mixture.The combined extract was diluted to 100 mL,subjected to hydrophilic-lipophilic balance(HLB)solid-phase extraction,and then evaporated to near dryness under nitrogen before analysis.Separation was achieved using a BEH C_(18)ultra performance liquid chromatography column with 0.1%formic acid in methanol as the mobile phase.The detection method employed multi-reaction monitoring with positive and negative ion switching.Results The method demonstrated excellent linearity in quantifying the 48 kinds of antibiotics,the determination coefficients were all greater than or equal to 0.9922.Limits of detection of method ranged from 0.02to 0.47μg/kg,while limits of detection ranged from 0.07 to 1.56μg/kg.Recovery rates at 3 spiked levels ranged from 69.2%to 120.3%,with relative standard deviations ranging from 1.0%to 14.3%.The detection results of the actual sample were also consistent with the results of the standard method.Conclusion This method is characterized by its simplicity,speed,sensitivity,and reliability,rendering it suitable for the concurrent detection of 48 kinds of antibiotics spanning 5 categories in fish meat.