高效液相色谱-串联质谱法检测鱼肉中5种头孢类药物

Simultaneous Determination of 5 Cephalosporins in Fish by High Performance Liquid Chromatography-Tandem Mass Spectrometry

建立高效液相色谱-串联质谱法同时检测鱼肉中5种头孢类抗生素(头孢氨苄、头孢洛宁、头孢匹林、头孢喹肟和头孢唑啉)残留的检测方法。鱼肉基质经乙腈提取,Prime HLB(6 m L,200 mg)固相萃取小柱净化。采用AB-4500 Qtrap液相色谱-串联质谱仪,Accucore C18 RP-MS色谱柱(2.1 mm×100 mm,2.7 μm),以乙腈和体积分数0.1%的乙酸水溶液为流动相进行梯度洗脱,流速0.4 m L/min,柱温30℃。采用电喷雾离子源,正离子扫描,多反应监测模式,外标法定量。结果表明:5种头孢类药物在0~75 μg/kg范围内线性良好,且在8 min内完全分离;5种头孢类药物的检出限均为0.5 μg/kg,定量限为2.0 μg/kg;在10、25、40 μg/kg加标水平下,5种头孢类药物的批内平均加标回收率为81.0%~111.0%,相对标准偏差为0.41%~10.50%,批间平均加标回收率为84.7%~106.0%,相对标准偏差为0.88%~8.71%。

A high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） method was developed for the determination of five cephalosporins in fish （cephalexin, cefalonium, cephapirin, cefquinome and cefazolin）. Samples were extracted with acetonitrile and then the extract was cleaned up with a Prime HLB solid-phase extraction （SPE） cartridge prior to being analyzed using an AB-4500 Qtrap LC-MS/MS system. The chromatographic separation was performed on an Accucore C18 RP-MS column （2.1 mm ×100 mm, 2.7 Jam） by gradient elution using a mobile phase made up of acetonitrile and 0.1% acetic acid in water at a 0.4 mL/min flow rate. The column temperature was set at 30 ℃. The analytes were detected using multiple-reaction monitoring （MRM） in the positive electrospray ionization mode and quantified by the external standard method. Complete separation was achieved within 8 min. The calibration curves for all five analytes were linear over the concentration range from 0 to 75 μg/kg. The limit of detection for all analytes was 0.5 μg/kg, and the limit of quantification was 2.0 p.g/kg. At spiked levels of 10, 25 and 40 μg/kg, the intra-assay recoveries of all five cephalosporins were 81.0%-111.0% with relative standard deviation （RSD） of 0.41%-10.50%, and the inter-assay recoveries were 84.7%- 106.0% with RSD of 0.88%-8.71%.