高效液相色谱-串联质谱法测定畜禽肉中9种青霉素的残留量

Determination of Nine Penicillin Residues in Livestock and Poultry Meat by High Performance Liquid Chromatography-Tandem Mass Spectrometry

文章建立了高效液相色谱-串联质谱(High performance liquid chromatography-tandem mass spectrometry,HPLC-MS/MS)法同时测定畜禽肉中9种青霉素药物残留的检测方法。样品采用磷酸盐缓冲溶液(pH5.5)提取,C_(18)固相萃取柱净化,将样品引入HPLC-MS/MS进行分析。以添加0.2%甲酸的乙腈-水(体积比)为流动相进行梯度洗脱,经Agilent ZORBAX Eclipse Plus-C_(18)(2.1 mm×150 mm,3.5μm)色谱柱分离,以电喷雾正离子模式扫描,在多反应监测(Multiple reactions monitoring,MRM)模式下采集,基质匹配标准溶液通过外标法定量。结果表明,9种目标物可在11 min内实现有效分离;青霉素G和萘夫西林在0.4~10.0 ng/mL浓度范围内线性关系良好,其余7种青霉素在2.0~50.0 ng/mL浓度范围内均具有良好的线性关系,相关系数r均大于0.9990;检出限(S/N≥3)为0.4~1.0μg/kg,定量限(S/N≥10)为0.8~2.0μg/kg;以牛肉和鸡肉为基质进行4水平加标试验(n=6),9种青霉素的回收率为70.8%~106.3%,相对标准偏差为2.6%~7.6%。该方法简单快速、灵敏度高、重现性好,适用于畜禽肉中9种青霉素残留的同时检测。

A method for simultaneous determination of 9 penicillin antibiotics in livestock and poultry meat by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was established.The samples were extracted by phosphate buffered solution(pH5.5),purified by C_(18) solid phase extraction column.Then the samples were analysed by HPLC-MS/MS.The Agilent ZORBAX Eclipse Plus-C_(18)(2.1 mm×150 mm,3.5μm)was used as stationary phase,while a mixture of 0.2%formic acid acetonitrile and 0.2%formic acid(V/V)aqueous solution at different volume ratios was used as mobile phase for gradient elution.Multiple reaction monitoring(MRM)mode via positive electrospray ionization was adopted in MS analysis,while the results were quantified with matrix matched standard solution by external standard method.As shown by the results,9 penicillins could be effectively separated within 11 min.The line concentration range of penicillin G and nafcillin was from 0.4 to 10.0 ng/mL,and the other seven penicillins had a good linear relationship in the concentration range of 2.0 to 50.0 ng/mL,while the correlation coefficients r were all greater than 0.9990.The limits of detection(3 S/N)was from 0.4 to 1.0μg/kg,while the limits of quantitation(10 S/N)was between 0.8 and 2.0μg/kg.Test for recovery was made on blank samples of beef and chicken at the 4 concentration levels by standard addition method,giving results in the range of 70.8%–106.3%,with RSDs(n=6)of the determined values in the range of 2.6%–7.6%.The method which had the superiority of simplicity,speediness,sensitivity and reproducibility,was suitable for the detection of nine penicillins in livestock and poultry meat simultaneously.