高选择性MRM3液质联用技术检测复杂动物性食品中β-受体激动剂残留

Determination of β-agonists residue in complex animal derived food by LC-MS/MS/MS in high selectivity MRM3

关键词:  MRM_³  液质联用技术  复杂动物性食品  β-受体激动剂  残留

Key words:  MRM_³  LC-MS/MS/MS  complex animal derived food  β-agonists  residue

摘要:

建立了高选择性MRM_³液质联用技术检测猪肝脏等复杂动物性食品中克仑特罗、莱克多巴胺和沙丁胺醇三种β-受体激动剂残留的新方法。猪肝脏样品经酶解后用乙酸乙酯和叔丁基甲醚萃取,MCX固相萃取柱净化,0.1%甲酸乙腈溶液和0.1%甲酸溶液为流动相,Phenomenex F5(50×3.0mm,2.6μm)色谱柱进行梯度洗脱,在电喷雾正离子源下,采用三级质谱多反应监测模式(MRM_³)检测。结果表明：克仑特罗、莱克多巴胺和沙丁胺醇在系列浓度范围内均呈现良好线性关系,相关系数R_²均大于0.99,猪肝脏组织中的定量限均为0.5μg/kg。从0.5、2和10μg/kg三个添加浓度检测结果可以看出,三种药物的回收率范围为89.7%~102.1%,批内、批间相对标准偏差均小于20％。该方法稳定性好,选择性强,灵敏度高,定性定量更加准确。

Abstract:

A novel method based on LC-MS/MS/MS in high selectivity MRM_³ was established for the determination of clenbuterol、ractopamine and salbutamol residues in complex animal derived food. The pig liver sample was extracted by ethyl acetate and tert-butyl methyl ether after enzymolysis, and purified on a mixed-mode cation exchange solid phase extraction column(MCX). A F5 chromatography column was used with acetonitrile(0.1% formic acid) and water(0.1% formic acid) as mobile phases in a gradient elution. Ionization was performed using an electrospray positive ion source(ESI_⁺),and detection was achieved by MS/MS/MS in multiple reaction monitoring(MRM_³) mode. The linearity for clenbuterol、ractopamine and salbutamol was good between serial concentrations, and the correlation coefficient was R_²＞0.99. The limits of quantification of clenbuterol、ractopamine and salbutamol in pig liver tissue was 0.5μg/kg, The average recoveries from spiked pig liver tissues at three concentrations of 0.5, 2 and 10μg/kg ranged 89.7%~102.1%. The intra- and inter-batch variation coefficients were both less than 20%. This method was good stability, high sensitivity, qualitative and qualitative more accurately .