固相萃取-液相色谱串联质谱法测定尿中4-硝基苯代谢物

Determination of the metabolites of 4-nitrobenzene in urine using solid-phase extraction coupled with liquid chromatography tandem mass spectrometry

目的建立了尿中4-硝基苯代谢物4-硝基苯胺、4-硝基苯酚同时测定的高效液相色谱串联质谱方法。方法将10 ml样品过ENVI-18固相萃取柱富集,采用C18反相色谱柱分离,用甲醇和乙酸铵溶液为流动相进行梯度洗脱,质谱采用电喷雾正离子或负离子电离,多反应监测模式检测。结果 4-硝基苯胺、4-硝基苯酚分别在质量浓度为0.50μg/L^100.0μg/L和0.050μg/L^10.0μg/L时,具有良好的线性关系,相关系数(r)≥0.999 4,方法的检出限为0.002μg/L^0.02μg/L,回收率为87.2%~99.6%,相对标准偏差为2.6%~4.8%。结论此方法前处理简便、灵敏度高,具有良好的回收率和精密度。适用于尿中4-硝基苯代谢物即4-硝基苯胺、4-硝基苯酚的同时测定。

Objective A method of liquid chromatography tandem mass spectrometry（ LC-MS / MS） was developed for the simultaneous determination of the 4-nitrobenzene metabolites（ 4-nitrophenol and 4-nitroaniline） in urine. Methods 10 ml of samples were concentrated by an ENVI-18 column. The separation was performed on a reverse-phase chromatographic C18 column with methanol and ammonium acetate as the mobile phase for gradient elution. The MS / MS detection was performed under positive or negative electrospray ionization（ ESI） in multiple reaction monitoring（ MRM） mode. Results The good linearity was obtained when the concentrations of 4-nitrophenol and 4-nitroaniline among 0. 050 μg / L-10. 0 μg / L and 0. 50 μg / L-100. 0 μg / L,and the correlation coefficients（ r） were ≥0. 999 4. The limits of detection were within 0. 002 μg / L-0. 02 μg / L,the recoveries were within 87. 2%-99. 6%,and relative standard deviation was among 2. 6%-4. 8%. Conclusion The method was simple,feasible and accurate with good recoverage and precision,and it could be applied to the simultaneous detection of 4-nitrophenol and 4-nitroaniline in urine.