超高效液相色谱-四极杆/静电场轨道阱质谱测定猪尿中环丙氨嗪及其代谢物残留量  

Determination of Residual Cyromazine and Its Metabolite Melamine in Pig Urine Samples Using Ultrahigh Performance Liquid Chromatography Coupled with Quadrupole-Orbitrap Mass Spectrometry

作　　者：罗秋红 韩颖 胡蓓 邓智伟 曾臻 万建春 占春瑞 LUO Qiu-hong;HAN Ying;HU Bei;DENG Zhi-wei;ZENG Zhen;WAN Jian-chun;ZHAN Chun-rui(Nanchang Customs Technical Center,Nanchang 330038,China)

机构地区：[1]南昌海关技术中心,江西南昌330038

出　　处：《分析测试学报》2021年第12期1784-1789,共6页Journal of Instrumental Analysis

摘　　要：建立了超高效液相色谱-四极杆/静电场轨道阱质谱(UHPLC-Quadrupole-Orbitrap MS)检测猪尿中环丙氨嗪及其代谢物三聚氰胺的方法。样品用1%三氯乙酸超声提取,调节pH<3.0,阳离子交换固相萃取柱净化氮气吹干后,95%乙腈水溶液复溶,经Syncronis HILIC色谱柱(100 mm×2.1 mm,1.7μm)分离,10 mmol/L乙酸铵水溶液-乙腈为流动相梯度洗脱,使用电喷雾离子源在平行反应监测模式(PRM)下检测。环丙氨嗪采用外标法定量,三聚氰胺采用内标法定量。对比了PRM模式与三重四极杆质谱的多反应监测模式(MRM),结果显示两者具有相当的定量能力,但PRM模式根据目标化合物及其二级子离子的精确质量数和丰度比进行定性分析,具有较强的定性优势。最佳条件下,2种目标物均在0.2~50.0 ng/mL范围内线性关系良好(r2>0.99),定量下限均为1.0μg/kg。在1.0、2.0、150.0μg/kg加标水平下,回收率为78.0%~105%,相对标准偏差(RSD)不大于11%。该方法可同时准确测定猪尿中环丙氨嗪和三聚氰胺的残留量。An ultrahigh performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry(UHPLC-Quadrupole-Orbitrap MS)was developed for the determination of cyromazine and its metabolite melamine in pig urine samples.The pig urine sample(2.0 g)was ultrasonically extracted with 1%trichloroacetic acid under the pH value of the extraction mixture lower than 3,followed by purification with a mixed cation solid phase extraction column(MCX),then the eluent from the MCX column was blow dried under nitrogen flow,and finally reconstituted with 95%acetonitrile aqueous solution.The reconstituted solution 10 times diluted was injected into the UHPLCQuadrupole-Orbitrap system using a Syncronis HILIC column(100 mm×2.1 mm,1.7μm)at 35℃,with 10 mmol/L ammonium acetate and acetonitrile as the mobile phases.The mass spectrometer was operated in positive electrospray(ESI+)and parallel reaction monitoring(PRM)mode.Qualitative analysis was carried out according to the accurate masses of each target compound,its fragment ions,and the abundance ratios of the fragment ions.The external standard method and internal standard method were used for the quantification of cyromazine and melamine,respectively.Method validation showed that there were good linear relationships for the two targets in the range of 0.2-50.0 ng/mL,with their correlation coefficients(r2)larger than 0.99.At three spiked levels of 1.0,2.0 and 150.0μg/kg,the recoveries for cyromazine and melamine in pig urine sample ranged from 78.0%to 105%,with relative standard deviations(RSD)no more than 11%.The same spiked samples were used for comparison between PRM of orbitrap and multiple reaction monitoring(MRM)mode of triple quadrupole.The results obtained exhibited no significant difference,indicating that both methods have good quantitative ability for determination of cyromazine and melamine in pig urine sample.With good recovery,dynamic range and quantification accuracy,the developed method could be applied to the simultaneous qualitative and quantitative analys

关 键 词：猪尿 环丙氨嗪 三聚氰胺 超高效液相色谱-四极杆/静电场轨道阱质谱 

分 类 号：O657.63[理学—分析化学] TS207.3[理学—化学]