超高效液相色谱法同时测定尿中甲苯、二甲苯、苯乙烯代谢产物

Simultaneous determination of the metabolites of methylbenzene,xylene and styrene in urine by ultra performance liquid chromatography

目的建立超高效液相色谱法同时测定尿中苯乙烯的代谢产物苯乙醇酸(MA)和苯乙醛酸(PGA)、甲苯的代谢产物马尿酸(HA)、二甲苯的代谢产物甲基马尿酸(MHA)(含3种同分异构体)的方法。方法尿液样本用HCl酸化后,经乙酸乙酯提取,吹氮浓缩,用流动相稀释后直接进样,采用20 mmol/L乙酸铵-甲醇为流动相,波长235 nm,流速0.25 ml/min,Waters ACQUITY UPLC~CSH^(TM)C_(18)柱分离,保留时间定性,峰面积定量。结果标准曲线为2μg/ml^400μg/ml,相关系数均>0.999 9,相对标准偏差为0.17%~6.51%,回收率为83.2%~107.0%,检出限MA为0.009μg/ml^0.13μg/ml,分析时间为6 min。结论该方法操作简便,快速,准确,灵敏,适合尿中MA、PGA、HA和MHA的测定。可用于日常接触苯乙烯、甲苯、二甲苯等危害因素工作人员的职业健康监测,具有一定的实际应用价值。

Objective To establish the ultra performance liquid chromatography method for simultaneous determination of metabolites of methylbenzene（ hippuric acid）,xylene（ methyl hippuric acid,including three isomers） and styrene（ mandelic acid and phenylglyoxylic acid） in urine. Methods The urine samples were acidfied by hydrochloric acid,extracted with ethyl acetate,concentrated by nitrogen,and diluted and sampled by mobile phase 20 mmol/L ammonium acetate and methanol. The wavelength was 235 mm and the velocity was 0. 25 ml/min. Waters ACQUITY UPLCCSHTMC18 collumn was used as separation column. Qualitative analysis was based on retention time and quantitative analysis was based on the peak areas.Results The standard curve was in the range of 2 g/ml-400 g/ml,the correlation coefficient was greater than 0. 999 9. The average recoveries were within 83. 2%-107. 0% with relative standard deviations of 0. 17%-6. 51%. The detection limits of mandelic acid（ MA） were within 0. 009 μg/ml-0. 13 μg/ml. The metabolites in urine were separated and detected within 6min. Conclusion The method is simple,rapid,reliable,accurate and sensitive and it is suitable for the determination of hippuric acid,methyl hippuric acid,mandelic acid and phenylglyoxylic acid in urine. The method may be used for monitoring the health of the staff contacting with the methylbenzene,xylene,styrene and has practical value.