Department of Environmental Health, Harvard School of Public Health, 677 Huntington Avenue, Boston, MA 02115, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Sep 1;904:73-80. doi: 10.1016/j.jchromb.2012.07.022. Epub 2012 Jul 31.
Phthalates and bisphenol A are environmental endocrine-disrupting chemicals used widely in common consumer products. There is increasing concern about human exposure to phthalates and bisphenol A due to the potential adverse effects related to the anti-androgenic activity of phthalates and estrogenic activity of bisphenol A. In assessing environmental exposure to phthalates and bisphenol A, it is essential to have a validated analytical method that can quantify trace concentrations of phthalate metabolites and bisphenol A in humans. In this study, we developed and validated an accurate, sensitive, and robust LC-MS/MS method to simultaneously quantify 5 phthalate monoester metabolites, including mono-methyl phthalate, mono-ethyl phthalate, mono-butyl phthalate, mono-benzyl phthalate, mono-2-ethylhexyl phthalate, and bisphenol A in human urine. In this method, the phthalate metabolites and bisphenol A, along with their isotope labeled internal standards, were extracted from 200 μl of human urine using automated off-line solid phase extraction. The analytes were quantitatively determined using LC-MS/MS operated in negative electrospray ionization multiple reaction-monitoring mode. The limit of quantification was 0.3 ng/ml for mono-methyl phthalate, mono-ethyl phthalate, mono-benzyl phthalate and bisphenol A, and 1 ng/ml for mono-butyl phthalate and mono-2-ethylhexyl phthalate. The precision and accuracy were well within the acceptable 15% range. This validated method has been used successfully in assessing exposure to phthalates and bisphenol A in humans.
邻苯二甲酸酯和双酚 A 是广泛应用于常见消费产品的环境内分泌干扰化学物质。由于邻苯二甲酸酯的抗雄激素活性和双酚 A 的雌激素活性，人们越来越关注人类接触邻苯二甲酸酯和双酚 A 所带来的潜在不利影响。在评估环境中邻苯二甲酸酯和双酚 A 的暴露情况时，必须要有一种经过验证的分析方法，能够定量检测人体中痕量邻苯二甲酸酯代谢物和双酚 A 的浓度。在这项研究中，我们开发并验证了一种准确、灵敏、稳健的 LC-MS/MS 方法，能够同时定量检测人体尿液中的 5 种邻苯二甲酸单酯代谢物，包括单甲基邻苯二甲酸、单乙基邻苯二甲酸、单丁基邻苯二甲酸、单苄基邻苯二甲酸、单 2-乙基己基邻苯二甲酸和双酚 A。在该方法中，邻苯二甲酸代谢物和双酚 A 及其同位素标记的内标物，通过自动离线固相萃取从 200μl 人尿中提取。使用 LC-MS/MS 在负电喷雾电离多反应监测模式下进行定量分析。单甲基邻苯二甲酸、单乙基邻苯二甲酸、单苄基邻苯二甲酸和双酚 A 的定量下限为 0.3ng/ml，单丁基邻苯二甲酸和单 2-乙基己基邻苯二甲酸的定量下限为 1ng/ml。精密度和准确度均在可接受的 15%范围内。该验证方法已成功用于评估人体接触邻苯二甲酸酯和双酚 A 的情况。
