超高效液相色谱-串联质谱法检测猪尿中20种β-受体激动剂残留

Determination of β-agonists in pig urine by UPLC-MS/MS

关键词:  猪尿  β-受体激动剂  残留  超高效液相色谱-串联质谱

Key words:  pig urine  β-agonists  residue  UPLC-MS/MS

摘要:

建立了猪尿中吡布特罗、西马特罗、特步他林等20种β-受体激动剂残留检测的超高效液相色谱-串联质谱方法。猪尿样品经酶解后，调节pH值至碱性，用叔丁醇和叔丁基甲醚（6 4，V/V）萃取，MCX柱净化，以0.1%甲酸乙腈溶液和0.1%甲酸水溶液为流动相进行梯度洗脱。同位素内标法和外标法定量。20种β-受体激动剂在系列浓度范围内呈现良好线性关系，相关系数R2均大于0.99；20种药物在猪尿中的检测限为0.25μg/L，定量限为0.5μg/L。从0.5、1和5μg/L三个添加浓度检测结果可以看出，20种药物的回收率为75.7%～110.7%，批内批间相对标准偏差均小于20％。该方法具有简便快捷、灵敏度高、定性准确等特点。

Abstract:

A UPLC-MS/MS method was established for the determination of Pirbuterol、Cimaterol、Terbutaline、Zilpaterol、Salbutamol、Cimbuterol、Clencyclohexerol、Clenproperol 、Hydroxymethyl clenbuterol、Clorprenaline、Ractopamine、Clenhexerol、Clenbuterol、Tulobuterol、Formoterol、Brombuterol、Clenpenterol、Bambuterol、Mabuterol and Mapenterol residue in pig urine. After enzymolysis, the solution was adjusted to base by pH, and then was extracted by tertiary butanol and tert-butyl methyl ether(6 4,V/V). The separation of β-agonists was performed with BEH C18 column and the gradient elutinon solvent of acetonitrile(0.1% formic acid) and water(0.1% formic acid) at a flow rate of 0.3mL/min. The method was quantified by internal standard method and external method, The calibration curves were good linear between 2～50μg/L,the correlation coefficient R2＞0.99. The limits of detection of 20 β-agonists in pig urine were 0.25μg/L , and the limit of quantification was 0.5μg/L. The average recoveries from spiked urine at three concentrations of 0.5,1,and 5μg/L ranged 75.7%～110.7%. The intra- and inter-batch variation coefficients were both less than 20%. This method was simple and rapid, and it was high sensitivity and qualitative accurately .