UPLC-MS/MS法测定猪肉中万古霉素与去甲万古霉素

摘要

建立了快速、准确、灵敏的猪肉中万古霉素和去甲万古霉素的超高效液相色谱串联三重四极杆质谱(UPLC-MS/MS)检测方法.样品经0.1％甲酸-乙腈(7:3,体积比)提取,正己烷(乙腈饱和)脱脂后,以LC-C1s固相萃取小柱净化,采用乙腈-0.1％甲酸为流动相进行梯度洗脱,串联质谱ESI正模式电离,多反应监测(MRM)模式检测,以保留时间和子离子比定性,外标法定量.结果表明,万古霉素和去甲万古霉素可在3 min内分离完全,在1～50 μg/L范围内线性关系良好,相关系数均大于0.99,方法检出限(S/N=3)分另为0.5 μg/kg和0.3μg/kg;加标水平为1～20 μg/kg时(n=6),平均回收率为80％～88％,日内相对标准偏差不高于13.4％,日间相对标准偏差不高于12.5％.方法准确、灵敏,适用于猪肉中万古霉素和去甲万古霉素等糖肽类抗生素的测定.%A simple, rapid and sensitive method was developed for the simultaneous determination of vancomycin and norvancomycin in pork by ultra-high performance liquid chromatography - tandem mass spectrometry (UPLC - MS/MS). The sample was supersonically extracted with a mixture of 0. 1 % formic acid - acetonitrile (7: 3), and cleaned up with hexane ( acetonitrile saturated) and a LC - C18 solid phase extraction cartridge. The separation of two targets was carried out on a Waters UPLC BEH C18 column by gradient elution with 0. 1% formic acid - acetonitrile as mobile phase. With positive electrospray ionization (ESI+) and multiple reaction monitoring (MRM), the two com pounds were identified with retention time and ion ratio. The results showed that the two compounds displayed excellent peak shapes within 3 min with correlation coefficients more than 0. 99. The limits of detection (LOD, S/N=3) of vancomycin and norvancomycin were 0. 5 μg/kg and 0. 3 μg/kg, respectively. The average recoveries were between 80% and 88% for the target compounds at spiked levels of 1 - 20 μg/kg. The intra-day RSDs were no more than 13. 4%, and the inter-day RSDs were no more than 12. 5% . The method was accurate and sensitive, and was suitable for the quantitative and qualitative analyses of glycopeptide antibiotics in pork sample.