固相萃取-液相色谱/串联四级杆质谱法测定尿中双酚A类物质

Testing bisphenol A analogs in human urine by solid-phase extraction liquid chromatography and tandem quadrupole mass-spectrometry

目的建立高效液相色谱/串联四级杆质谱法同时测定尿中4种双酚A类物质的方法。方法尿样用β-葡糖苷酸酶/硫酸酯酶水解后,经瓦里安Bond Elute Plexa固相萃取小柱浓缩净化,以乙腈-水为流动相进行梯度洗脱,采用液相色谱-质谱/质谱电喷雾负离子电离模式和多反应监测扫描模式,以双酚A-d16和四溴双酚A-d10为内标物进行定量分析。结果尿中双酚F、双酚A、四氯双酚A和四溴双酚A的线性范围均为0.50～100.00μg/L,线性相关系数分别为0.998 6、0.999 1、0.999 6和0.999 7,检出限分别为0.21、0.25、0.01和0.14μg/L,加标回收率为69.2%～99.9%;批内和批间相对标准偏差分别为5.5%～22.8%和2.4%～14.4%;样品在室温下至少可以保存7 d。对38份儿童尿样及20份医务工作者尿样进行检测,前者双酚F、双酚A、四氯双酚A和四溴双酚A检出率分别为94.7%、94.7%、0.0%和34.2%,后者分别为100.0%、100.0%、5.0%和75.0%。结论本方法简便,灵敏度高,选择性好,可作为检测尿中4种双酚A类物质的有效方法。

Objective To develop a method for simultaneous detection of 4 kinds of bisphenol A analogs in human urine by high performance liquid chromatography and tandem quadrupole mass-spectrometry. Methods The urine samples were extracted by solid phase extraction（ Bond Elut Plexa） after hydrolyzed using β-glucuronidase and sulfatase. The gradient elution was carried out with acetonitrile-water. The bisphenol A-d16 and tetrabromobisphenol A-d10 were used as internal standard substances to conduct the quantitative analysis using the combined mode of liquid chromatography-mass spectrometry and mass spectrometry with the negative electrospray ionization,and the multiple reaction monitoring scan mode. Results The linear range of all the 4 kinds of bisphenol A analogs（ Bisphenol F, Bishpenol A,Tetrachlorobisphenol A and Tetrabromobisphenol A） in urine was found to be 0. 50-100. 00 μg / L, and the linear correlation coefficients were 0. 998 6,0. 999 1,0. 999 6 and 0. 999 7,respectively. The limits of detection were 0. 21,0. 25,0. 01 and 0. 14 μg / L. The recovery rate was 69. 2%-99. 9%. The relative standard deviations of within-run and between-run precisions were 5. 5%-22. 8% and 2. 4%-14. 4%,respectively. The samples could be kept for at least 7 days at room temperature. The method was applied to detect urine samples of 38 children and 20 medical workers and the results were statistically analyzed. The detection rates of bisphenol A, bisphenol F, tetrachlorobisphenol A and tetrabromobisphenol A for children group were 94. 7%,94. 7%,0. 0% and 34. 2%,respectively,and those of the medical workers group were 100. 0%,100. 0%,5. 0% and 75. 0%,respectively. Conclusion The method is effective for the detection of 4 kinds of bisphenol A analogs in human urine with simple operation,high sensitivity and good selectivity.