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

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一串联质谱方法。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1．2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％，批内批间相对杯准偏差均小于20％。该方法具有简便快捷、灵敏度高、定性准确等特点。

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 wasextracted 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 0.3 mlMmin. The method was quantified by internal standard method and external method, The calibration curves were good linear between 2 -50 μg/L, the correlation coefficient R2 〉 0. 99. The limits of detection of 20 β- agonists in pig urine were 0.25 μg/L , and the limit of quantification was 0.5 μg/L. The average recoveries from spiked urine at three concentrations of 0.5, land 5 μg/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.