超高效液相色谱串联质谱法检测鸡蛋中残留的氨基糖苷类药物

目的:建立鸡蛋中5种氨基糖苷类药物残留的UPLC-MS/MS检测方法。方法:样品经磷酸盐缓冲液提取,调节pH后,过C₁₈固相萃取小柱净化,氮气吹干后七氟丁酸溶液复溶,进行UPLC-MS/MS分析。色谱柱:BEH C₁₈(50 mm×2.1 mm,1.7 μm);流动相:20 mmol·L^-1七氟丁酸乙腈(A) -20 mmol·L^-1七氟丁酸水溶液(B),梯度洗脱(0~2 min,4% A线性变化至30% A;2~4 min,30% A线性变化至60% A;4~5 min,60% A线性变化至96% A;5~5.5 min,96% A线性变化至4% A;5.5~7.0 min,维持4% A);流速:0.3 mL·min^-1;柱温:30℃;进样量:10 μL。结果:5种氨基糖苷类药物质量浓度在200~1000 ng·mL^-1范围内呈现良好的线性关系,相关系数r均大于0.995;添加500 ng·g^-1时回收率范围为67.9%~79.4%,批内RSD范围为1.03%~4.08%,批间RSD范围为7.77%~16.50%;方法检测限为5 μg·kg^-1,定量限为20 μg·kg^-1。20份实际鸡蛋样品中均未检出5种氨基糖苷类药物。结论:经方法学验证,本方法可用于鸡蛋中氨基糖苷类药物的检测。

Objective:To develop a UPLC-MS/MS analysis method for the determination of 5 aminoglycosides in eggs.Methods:The samples were extracted with phosphate buffer solution.After adjusting the pH value, the solution was purified with C₁₈ solid phase extraction column,dried by nitrogen and reconstituted with seven fluorine butyric acid solution.UPLC-MS/MS analysis was then performed.The separation was carried out on a Waters BEH C₁₈(50 mm×2.1 mm,1.7 μm)column with gradient elution(0-2 min,4%A linear change to 30%A;2-4 min,30%A linear change to 60%A;4-5 min,60%A linear change to 96%A;5-5.5 min,96%A linear change to 4%A;5.5-7.0 min,4%A maintained)using acetonitrile with 20 mmol·L^-1 seven fluorine butyric acid(A) -20 mmol·L^-1 seven fluorine butyric acid(B).The flow rate was 0.3 mL·min^-1,the column temperature 30℃,and the injection volume 10 μL.Results:Excellent linear relationship was obtained from 200 ng·mL^-1 to 1000 ng·mL^-1 for the five aminoglycosides(correlation coefficient r≥0.995),the recoveries ranged from 67.9% to 79.4% at 500 ng·g^-1,and intra-and inter-batch variation coefficients were 1.03%-4.08%,and 7.77%-16.50%,respectively.The detection limit was 5 μg·kg^-1,and the limit of quantification was 20 μg·kg^-1. The five aminoglycosides were not detected in any of the 20 samples.Conclusion:The methodology validation showed that the established method is specific,simple,accurate,and rapid,which can be used to determine aminoglycosides in eggs.