固相萃取-液相色谱-串联质谱法测定海参中62种兽药残留

Determination of 62 kinds of veterinary drug residues in sea cucumber by solid phase extraction with liquid chromatography-tandem mass spectrometry

目的建立固相萃取结合高效液相色谱-串联质谱同时测定海参中62种药物的检测方法。方法样品加水分散后经1.0%甲酸乙腈溶液提取,PEP固相萃取柱净化后,采用WatersX-Bridge-C18色谱柱分离、0.2%甲酸水溶液-甲醇为流动相进行梯度洗脱,使用选择离子监测模式检测。结果62种兽药在0.50~100.00ng/m L范围内呈现出良好的线性关系,线性相关系数r^(2)在0.9953~1.0000范围内。方法检出限为0.25~2.50μg/kg;在3个不同浓度添加水平下,62种兽药的平均回收率在70.3%~119.0%,批内和批间的相对标准偏差(n=5)为0.13%~14.90%,均小于15.00%。将该方法应用于30批次海参样品的检测,其中4批次样品的喹诺酮检测结果阳性,检出量为5.30~28.00μg/kg,与国家标准方法的检测结果相比,该方法对于本次喹诺酮药物筛查的准确率为100%。结论该方法灵敏度高,准确度和精密度良好,满足我国兽药残留检测要求,可适用于海参中62种兽药多残留的定性定量分析检测。

Objective To establish a method for simultaneous determination of 62 kinds of veterinary drugs in sea cucumber by solid phase extraction combined with liquid chromatography tandem mass spectrometry.Methods The sample was extracted with 1.0%formic acid acetonitrile after dispersed with water.After purification by PEP solid-phase extraction column,62 kinds of veterinary drugs were separated using Waters X-Bridge-C18 column with 0.2%formic acid aqueous solution and methanol as mobile phase.Detection was performed using selective ion monitoring mode.Results The results showed that 62 kinds of veterinary drugs exhibited good linearity in the range of 0.50–100.00 ng/mL,with a correlation coefficient r^(2) in the range of 0.9953~1.0000.The detection limit of the method is 0.25–2.50µg/kg.Recovery test with 3 different addition contents showed the results in the range of 70.3%-119.0%,and all the relative standard deviation RSD(n=5)were 0.13%-14.90%,less than 15.00%.At last the method was applied to detection of 30 sea cucumber samples,among which 4 samples contained quinolone with the amount range from 5.30–28.00µg/kg.Compared with the results of national standard method,the accuracy of this method for quinolone drug screening was 100%in this study.Conclusion This method has high sensitivity,good accuracy and precision,which meets the requirements of China’s veterinary drug residue detection,and can be applied to the qualitative and quantitative detection of 62 kinds of veterinary drug residues in sea cucumbers.