UPLC-MS/MS检测牛奶中19种β-内酰胺类药物残留的两种前处理方法对比试验

Comparative test of two pretreatment methods for detection of 19β-lactam drugs residues in milk by UPLC-MS/MS

关键词:  牛奶  β-内酰胺类  残留  前处理

Key words:  milk  β -lactam  residual  pretreatment

摘要:

对比用超滤离心管和HLB-SPE柱两种不同前处理方法检测牛奶中19种β-内酰胺类药物残留的试验研究，确立一种简单、快捷，回收效果好的前处理方法。使用19种药物的混合标准溶液为外标，样品经沉淀蛋白，离心后同时用HLB-SPE柱和超滤离心管净化、富集处理，在Waters Acquity UPLCTM BEH C18柱(100 mm × 2.1 mm,1.7 μm)中分离，以0.1%甲酸水溶液/乙腈为流动相，经梯度洗脱将19种β-内酰胺类抗生素完全分离；采用正离子扫描方式确定丰度比最高的两对离子作为监测离子，进行MRM模式定性定量分析。结果表明：上述19种药物的检测限(LOD)为0.1～1.0 μg/kg，定量限(LOQ)为0.3～3.0 μg/kg，在10.0～100.0 μg/kg线性范围内，相关系数r均大于0.99，且用超滤离心管对牛奶进行净化浓缩处理比HLB柱的回收效果好，回收率均在60.7%～96.6%。超滤离心管的前处理方法简单，试剂消耗少，测定时间短，回收率好，结果准确可靠，可成功应用于牛奶中β-内酰胺类药物的残留检测。

Abstract:

This study compared ultrafiltration centrifuge tube with HLB-SPE column in two different pretreatment methods of detecting 19 β-lactam drugs residues in milk,to establish a simple, fast, good recovery pretreatment method.The samples were separated on an Acquity UPLCTM BEH C18 column (100 mm×2.1 mm,1.7 μm) after centrifugation with an UPLCTM SPE column and an ultrafiltration centrifuge for purification and enrichment.19 β -lactam antibiotics were separated by gradient elution using 0.1% formic acid aqueous solution/acetonitrile as mobile phase.Positive ion scanning method was used to determine the two pairs of ions with the highest abundance ratio as monitoring ions, and qualitative and quantitative analysis was carried out by MRM mode.Results:The limits of detection (LOD) and quantification (LOQ) of the 19 drugs were 0.1～1.0 μg/kg and 0.3～3.0 μg/kg respectively. The correlation coefficients r were all greater than 0.99 within the linear range of 10.0～100.0 μg/kg.The purification and concentration of milk by ultrafiltration centrifuge was better than that by HLB column,and the recovery rate was 60.7%～96.6%.The pretreatment method of ultrafiltration centrifugal tube was simple,the reagent consumption was less,the determination time was short,the recovery rate was good,the result was accurate and reliable, and it can be successfully used for the detection of β -lactam drug residues in milk.