通过式固相萃取净化-超高效液相色谱-串联质谱法测定食用肉类和肝中的4种β-受体激动剂残留量

Determination of 4 β-agonists residues in meat and liver by filtration solid phase extraction combined with ultra performance liquid chromatography-tandem mass spectrometry

目的采用通过式固相萃取方法进行样品净化,建立测定食用肉和肝中4种β-受体激动剂(特布他林、沙丁胺醇、莱克多巴胺和克伦特罗)残留量的超高效液相色谱-串联质谱检测方法。方法样品37℃经β-盐酸葡萄糖醛苷酶/芳基硫酸酯酶酶解过夜,乙腈提取,通过式固相萃取净化,采用Waters BEH C_(18)(100 mm×2.1 mm,1.7μm)色谱柱为固定相,5 mmol/L乙酸铵溶液和0.05%甲酸的乙腈溶液为流动相,超高效液相色谱-串联质谱法(UPLCMS/MS)检测,同位素内标法定量。结果空白牛肉基质中4种β-受体激动剂在1.0μg/L~200μg/L内相关系数≥0.990,检出限为0.5μg/kg~1.0μg/kg,定量限为1.5μg/kg~3.0μg/kg。在25.0μg/kg、125μg/kg、250μg/kg 3个加标水平时,回收率在80.4%~101.0%,精密度在2.8%~7.1%。结论本方法样品前处理简单、灵敏度高、重现性好,可用于食用肉和肝样品中4种β-受体激动剂类兽药残留量的同时快速分析检测。

Objective To develop a method for determination of four β-agonists including terbutaline,salbutamol,ractopamine,and clenbuterol in edible meat and liver by filtration solid phase extraction combined with ultra-performance liquid chromatography-tandem mass spectrometry( UPLC-MS/MS). Methods The samples were firstly enzyme digested with β-glucosidase/arylsulfatase overnight at 37 ℃,extracted with acetonitrile,followed by cleaned-up with solid phase extraction cartridge,using Waters BEH C_(18)( 100 mm × 2. 1 mm,1. 7 μm) column as stationary phase and acetonitrile containing 0. 05%formic acid-5 mmol/L ammonium acetate aqueous solution as mobile phase,finally determined by UPLC-MS/MS and quantified by internal standard method. Results The correlation coefficients( r) of the four target compounds in blank substrate were ≥0. 990 in the range of 1. 0 μg/L-200 μg/L. The limits of detection( LOD) and quantification( LOQ) were within0. 5 μg/kg-1. 0 μg/kg and 1. 5 μg/kg-3. 0 μg/kg,respectively. At three spiked levels of 25. 0 μg/kg,125 μg/kg and250 μg/kg,recoveries ranged from 80. 4% to 101. 0% and the relative standard deviations( RSDs) were within 2. 8%-7. 1%. Conclusion This method is simple,reproducible,accurate and sensitive which could be applied to analysis of the4 β-agonists in meat and liver samples successfully.