超高效液相色谱-串联质谱检测鸡蛋、鸡肉和猪肉中酰胺醇类药物残留

Simultaneous determination of amphenicol residues in eggs, chicken and pork by liquid chromatography-tandem mass spectrometry

为了检测畜禽产品中酰胺醇类药物残留是否超标,采用超高效液相色谱-串联质谱法建立了同时检测鸡蛋、鸡肉和猪肉中氯霉素、甲砜霉素、氟苯尼考及其代谢产物氟苯尼考胺等酰胺醇类药物的分析方法。样品经乙腈∶氨水(98∶2,V/V)和乙酸乙酯提取、氮吹、CNWBOND Si柱净化、正己烷去脂后,经ACQUITY UPLC CSH C18(100 mm×2.1 mm,1.7μm)柱分离,以2 mmol/L乙酸铵水溶液和2 mmol/L乙酸铵乙腈溶液为流动相进行梯度洗脱,流速为0.3 mL/min,多反应监测模式(MRM)检测,分别以氯霉素-d5为氯霉素、甲砜霉素内标,以氟苯尼考-d3为氟苯尼考、氟苯尼考胺内标进行内标法定量。结果显示:4种药物在0.2~100 ng/mL范围内均成良好线性关系,相关系数均在0.99以上,方法检测限为0.1μg/kg,定量限为0.5μg/kg,在0.5~2.0μg/kg添加范围内,回收率范围为70.6%~112.3%,批内相对标准偏差为1.5%~12.7%,批间相对标准偏差为3.2%~14.5%。结果表明该方法灵敏度高,稳定性好,能满足同时检测畜禽产品中多种酰胺醇类药物残留的分析要求。

The residues of phenicols in livestock and poultry products directly affect consumers’health.An ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was established for simultaneous determination of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine residues in eggs,chicken and pork.Samples were extracted with acetonitrile-ammonium hydroxide(98:2,V/V)and ethyl acetate,and were purified with the CNWBOND Si SPE column,followed by removing fat using hexane.The analytes were separated on a ACQUITY UPLC CSH C18 chromatographic column(100 mm×2.1 mm,1.7μm)and they received gradient elution using water with 2 mmol/L of ammonium acetate and acetonitrile with 2 mmol/L of ammonium acetate as flowing phases at a flow rate of 0.3 mL/min.The analysis was carried out by the multiple reaction monitoring(MRM)mode using chloramphenicol-d5 as the internal standard of chloramphenicol and thiamphenicol,and using florfenicol-d3 as the internal standard of florfenicol and florfenicol amine.The calibration curves of the four analytes were in good linear relationship in a concentration range of 0.2-100 ng/mL,and the correlation coefficients(r)were over 0.99.The limits of detection of the four analytes were 0.1μg/kg,and the limits of quantification were 0.5μg/kg.The average recoveries of the four analytes ranged from 70.6%to 112.3%,with the corresponding intra-and inter-day variations(relative standard deviation,R.S.D.)being 1.5%to 12.7%and 3.2%to 14.5%,respectively.The present method was sensitive and reliable meeting the requirement for simultaneous determination of residues of various phenicols in livestock and poultry products.