液相色谱-串联质谱法测定尿液中7种有机磷酸酯代谢物

摘要

利用液相色谱-三重四极杆质谱(LC-MS/MS)联用技术,建立了人尿液中7种有机磷酸酯代谢产物-有机磷酸二酯的检测方法.针对不同理化性质的有机磷酸二酯,采用固相萃取技术进行富集净化,筛选出高效的固相萃取柱,并对其洗脱条件进行优化;同时,对流动相和质谱参数进行优化,获取用于各代谢产物定性与定量分析的特征离子对.研究结果表明,前处理采用Oasis WAX固相萃取柱富集、2 mL含5%氨水的甲醇和2 mL甲醇洗脱目标物,除磷酸二乙酯(DEP,17.8%~36.2%)外,其余目标化合物回收率均在60.5%~104.0%之间.在优化的液相色谱条件下,7种化合物可达到完全基线分离.7种有机磷酸二酯的检测限和定量限分别在0.005~0.2μg/L和0.02~0.5μg/L之间,日内和日间精密度结果(RSD≤15.4%)表明,本方法具有较好的稳定性和重现性.本方法用于普通人群尿液中有机磷酸二酯的检测,7种化合物在尿液中均有检出,总浓度在0.5~6.7μg/L之间.%A simple method was developed for simultaneous determination of seven urinary metabolites of organophosphate esters by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Based on different physical and chemical properties of these OPs metabolites, the solid phase extraction cartridges and the washing and eluting solvents were optimized in details. Furthermore, the mobile phase and mass spectrometric parameters were also investigated. The results showed that Oasis WAX cartridge was the best SPE column in this study, and 2 mL of NH3 ·H2 O (5% ) in methanol and 2 mL of methanol were chosen as the eluting solvents. The recoveries of six analytes were ranged from 60. 5% to 104. 0% , whereas DEP ranged from 17. 8% to 36. 2% . Seven analytes could be baseline separated from each other under the optimized chromatographic conditions. The limits of detection and quantification of seven analytes ranged from 0. 005 to 0. 2 μg / L and 0. 02 to 0. 5 μg / L, respectively. The standard deviations of response repeatability for intra-day and inter-day period were lower than 15. 4% . This method was finally applied to determination of metabolites of OPs from 10 urines from general population in Guangzhou city. The concentrations of total OPs metabolites in urine samples ranged from 0. 5 to 6. 7 μg / L.