HPLC-MS／MS测定尿液BPA、TCS和4-n-NP方法的建立及其应用

Establishment and Application of Determination Method for BPA, TCS, and 4-n-NP in Urine by HPLC-MS/MS

[目的]建立用高效液相色谱串联质谱(HPLC-MS/MS)测定人尿液中双酚A(bisphenol A,BPA)、三氯生(triclosan,TCS)与正壬基酚(nonylphenol,4-n-NP)含量的方法,分析上海地区育龄男子体内上述3种物质的暴露情况。[方法]建立BPA、TCS和4-n-NP的前处理方法和HPLC-MS/MS检测方法,观察所建方法的线性关系、日内及日间精密度及回收率,并应用所建方法对育龄男性志愿者尿液中BPA、TCS和4-n-NP进行定量分析。[结果]尿液中BPA与TCS在0.1~50 ng/m L范围内线性关系良好(r2=0.999,r2=0.998),4-n-NP在0.01~10 ng/m L范围内线性关系良好(r2=0.998),尿液中BPA、TCS和4-n-NP最低检测限(LOD)分别为0.1、0.1和0.01 ng/m L,固相萃取回收率分别为97.6%、76.9%和33.2%,日内及日间精密度均小于15%。该方法应用于研究200名上海育龄男性人群体内污染物暴露情况,所测育龄男子人群中BPA、TCS及4-n-NP的检出率分别为99%、96%和62.5%,含量几何均数分别为1.17、1.57和0.021 ng/m L,含量分别为<LOD^83.7 ng/m L,<LOD^83.8 ng/m L和<LOD^0.472 ng/m L。[结论]本研究建立了尿液中BPA、TCS与4-n-NP的HPLC-MS/MS检测方法,灵敏度高,受干扰因素少,结果准确可靠。上海市育龄男子尿液中BPA、TCS及4-n-NP有较高检出率。

[ Objective ] To develop a sensitive method based on high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） for detecting bisphenol A （BPA）, triclosan （TCS）, and nonylphenol （4-n-NP） in human urine, and to assess the exposure to these three chemicals in men of reproductive age in Shanghai. [ Methods ] Pretreatment method and HPLC-MS/MS detection method for BPA, TCS, and 4-n-NP were developed. Linearity recovery, and intra-and inter-batch relative standard deviations of the proposed method were assessed. The established HPLC-MS/MS method was applied in male volunteers of reproductive age for quantitative analysis on BPA, TCS, and 4-n-NP in urine samples. [ Results ] The BPA, TCS, and 4-n-NP in urine were linear over the range of 0.1-50 ng/mL （r2=0.999）, 0.1-50 ng/mL （r2=0.998）, and 0.01-10ng/mL （r2=0.998）, respectively; the limits of detection （LOD） were 0.1, 0.1, and 0.01 ng/mL, respectively; the recoveries were 97.6%, 76.9%, and 33.2%, respectively; and all the relative standards of intra and inter batch precisions were less than 15%. The detectable rates of BPA, TCS, and 4-n-NP in the urine samples of 200 men of reproductive age in Shanghai were 99%, 96%, and 62.5%, and the geometric means of concentrations were 1.17, 1.57, and 0.021 ng/mL, in the range of 〈LOD-83.7, 〈LOD-83.8, and 〈LOD-0.472 ng/mL, respectively. [ Conclusion ] The proposed HPLC-MS/MS method is sensitive, accurate, and specific for determination of BPA, TCS, and 4-n-NP in human urine. All the three environmental pollutants are highly detectable in the urine samples of men of reproductive age in Shanghai.