二维超高效液相色谱-三重四极杆/复合线性离子阱质谱联用快速测定全血和尿液中鱼藤酮

Rapid determination of rotenone in whole blood and urine by two-dimensional ultra performance liquid chromatography-triple quadrupole/linear ion trap mass spectrometry

建立了采用二维超高效液相色谱-三重四极杆/复合线性离子阱质谱联用快速测定全血和尿液中鱼藤酮的检测方法。尿液经水等量稀释后直接进样分析;全血用乙腈沉淀除蛋白质,上清液用水稀释后进样分析。样品中的鱼藤酮被Cyclone柱保留并去除大分子杂质,然后再被流动相洗入第1维Kinetex Biphenyl色谱柱(联苯基核壳柱)进行分离,再将含有鱼藤酮的流分切割至第2维Acquity BEH C_8色谱柱上进行分离,采用多离子监测触发的增强子离子(MRM-IDA-EPI)扫描方式检测,溶剂标准外标法定量。全血和尿液中的平均加标回收率分别为84.6%~94.3%和88.4%~95.7%,相对标准偏差为2.6%~7.3%和2.8%~6.8%(n=6),方法的检出限为0.2和0.03μg/L。该方法已成功应用于鱼藤酮中毒样品的检测。

The two-dimensional ultra performance liquid chromatography-triple quadrupole/linear ion trap mass spectrometry （2D-UPLC-QTRAP MS） method has been developed for the determination of rotenone in whole blood and urine. This method is based on the use of two columns of different stationary phases （ Kinetex Biphenyl and Acquity BEH C8） connected through two six-port two-position switching valves. Urine samples were diluted with equal quantity of water and then directly injected into the separation system. Blood samples were pre-pared by precipitation of proteins with acetonitrile, and then the supernatants were diluted with water and injected into the system. The samples were loaded onto the Cyclone column at high flow-rates, allowing the excluded proteinaceous material to flow through to waste. Then the retained rotenone was eluted into the Kinetex Biphenyl column, and the first dimension separa-tion was completed. The fraction containing rotenone was switched into a trap column （XBridge C8Direct Connect HP）. After the rotenone was retained completely by trap column, the valve switched it into the stream of the second dimension. The rotenone was separated on the Acquity BEH C8 column with gradient elution of mobile phases of acetonitrile-H2 O contai-ning 0. 2% （v/v） formic acid,detected by positive electrospray ionization tandem mass spec-trometry in the multiple reaction monitoring- information-dependent acquisition-enhanced product ion （MRM- IDA-EPI） mode, and quantified by solvent standard solutions. The average recoveries were 84. 6%-94. 3% and 88. 4 % -95. 7% for rotenone in blood and urine with relative standard deviations of 2. 6%-7. 3% and 2. 8 % -6. 8% （n = 6）. The limits of detection were 0. 2and 0. 03 μg/L for blood and urine,respectively. The method is sensitive,selective,and has been successfully applied to the detection of rotenone in the samples resulting in food poisoning.