同位素稀释-超高效液相色谱-串联质谱法测定禽类食品中利巴韦林和金刚烷胺类化合物残留量

Determination the residue of ribavirin and adamantanes in poultry food by isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry

目的建立同位素稀释-超高效液相色谱-串联质谱(UPLC-MS/MS)法检测禽类食品中利巴韦林和金刚烷胺类化合物(金刚烷胺、金刚烷甲胺、金刚烷乙胺、3,5-二甲基金刚胺)的残留量。方法样品经酶解,三氯乙酸沉淀蛋白,低温高速离心,上清液调节pH值后经PBA/PCX复合固相萃取柱净化,Agilent ZORBAX SB-Aq柱(3.0 mm×100 mm,1.8μm)分析利巴韦林,Waters BEH C18色谱柱(2.1 mm×100 mm,1.7μm)分析金刚烷胺类化合物,串联质谱测定,同位素内标法定量。结果利巴韦林在1.0~100 ng/mL、金刚烷胺类化合物在0.2~20 ng/mL范围内呈良好的线性关系,相关系数(r)均为0.999。利巴韦林的检出限和定量限分别为0.5和1.5μg/kg;金刚烷胺类化合物的检出限和定量限分别为0.1和0.3μg/kg。利巴韦林(1.5~15μg/kg)和金刚烷胺类化合物(0.3~3.0μg/kg)添加3个浓度的检测结果显示,利巴韦林的回收率为91.4%~103.7%,金刚烷胺类化合物的回收率为94.3%~108.2%,相对标准偏差(RSD)均小于10%。结论本方法具有简便快捷、灵敏度高、定性准确等特点。

Objective An isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was established for the determination of ribavirin and adamantanes(amantadine,adamantanemethylamine,rimantadine,memantine)in poultry food.Methods After enzymatic hydrolysis and precipitated by trichloroacetic acid,the samples were centrifuged by high-speed centrifuge at low temperature,and purified by PBA/PCX composite solid-phase extraction cartridge.The separation of ribavirin was performed on UPLC system with Agilent ZORBAX SB-Aq(3.0 mm×100 mm,1.8μm),adamantanes with Waters BEH C18 column(2.1 mm×100 mm,1.7μm)and determinated by tandem mass spectrometry.The method was quantified by internal standard.Results The calibration curves showed a good linearity between the peak ratio and the concentrations of 1.0-100 ng/mL(ribavirin)and 0.2-20 ng/mL(adamantanes)with r=0.999.The limit of detection of ribavirin was 0.5 and 0.1μg/kg for adamantanes.The limit of quantification of ribavirin was 1.5 and 0.3μg/kg for adamantanes.The mean recoveries of the ribavirin ranged from 91.4%-103.7%and ranged from 94.3%-108.2%for adamantanes spiked at three concentration levels(1.5-15 and 0.3-3.0μg/kg),with the relative standard deviations(RSD)less than 10%.Conclusion This method was simple,high sensitivity and accuracy.