超高效液相串联质谱法分析牛肉中残留的β-受体激动剂

Determination of residual β-agonists in beef by UPLC-MS /MS

目的：建立牛肉中9种β-受体激动剂的UPLC-MS/MS分析方法,为食品安全监测提供数据支撑。方法：样品用β-葡萄糖醛苷酶酶解4 h,高氯酸沉淀除杂,调节pH后,过MCX固相萃取小柱净化,氮气吹干后甲酸溶液复溶,进行UPLC-MS/MS分析。色谱柱：BEH C18（50 mm×2.1 mm,1.7μm）;流动相：0.1%甲酸乙腈溶液（A）-0.1%甲酸水溶液（B）,梯度洗脱（0~1 min,维持5%A;1~3 min,5%A线性变化至50%A;3~4 min,50%A线性变化至60%A;4~5 min,60%A线性变化至99%A;5~5.5 min,99%A线性变化至5%A;5.5~7.5 min,维持5%A）;流速：0.4 mL·min-1;柱温：30℃;进样量：10μL。结果：9种β-受体激动剂在0.5~10.0μg·kg-1添加浓度范围内线性关系良好,相关系数r≥0.9990,较高浓度时回收率范围为62.7%~107.9%,定量下限（LLOQ）规定为添加回收试验中S/N≥10时的最低添加浓度,结果均小于0.5μg·kg-1。结论：经方法学验证,本方法专属性好,简便、准确、快速,适用于牛肉中β-受体激动剂的检测。

Objective： To develop a UPLC - MS/MS analysis method for the determination of 9 beta - agonists, so as to provide data support for guaranteeing food safety. Methods： The samples were digested with fl - glucuronidase for 4 hours, and the impurities were precipitated with perchloric acid ; after adjusting the pH value, the solution was purified with MCX solid phase extraction column, dried by nitrogen and reconstituted with formic acid solution. UP- LC -MS/MS analysis was performed. The separation was carried out on a Wsters BEH C18（50 mm ×2. 1 mm,1.7μm） column using acetonitrile with 0. 1% formic acid （A） - 0. 1% formic acid solution （ B ） with gradient elution （0 - 1 min,5 % A maintained ; 1 - 3 min,5 % A linear change to 50% - A ; 3 - 4 min,50% A linear change to 60% A;4 -5 min,60% A linear change to 99% A;5 - 5.5 min,99% A linear change to 5% A;5.5 - 7.5 min,5% A maintained）. The flow rate was 0.4 mL · min-l , the column temperature 30 ℃ , the injection volume 10 μL. Results：Excellent linear relationship was obtained from the additive concentration of 0. 5 μg ~ kg-1 to 10. 0 μg · kg-1 for the nine β -agonists（ correlation coefficient r≥0. 9990） ,the recoveries ranged from 62.7% to 107.9% at the higher concentration ; the lower limit of quantification （ LLOQ）, defined as the lowest concentration given the sig- nal -to -noise≥10： 1, was less than 0.5 μg · kg-1. Conclusion：The methodology validation shows that the estab- lished method is specific, simple, accurate, and rapid,which can be used to determine β- agonists in beef.