Department of Analytical Chemistry, University of the Basque Country (UPV/EHU), Leioa, Basque Country, 48940, Spain.
Research Centre for Experimental Marine Biology and Biotechnology (PiE), University of the Basque Country (UPV/EHU), Basque Country, Plentzia, 48620, Spain.
Anal Bioanal Chem. 2022 Sep;414(23):6855-6869. doi: 10.1007/s00216-022-04250-w. Epub 2022 Jul 29.
In the present work, a target analysis method for simultaneously determining 24 diverse endocrine-disrupting compounds (EDCs) in urine (benzophenones, bisphenols, parabens, phthalates and antibacterials) was developed. The target analysis approach (including enzymatic hydrolysis, clean-up by solid-phase extraction and analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS)) was optimized, validated and applied to volunteers' samples, in which 67% of the target EDCs were quantified. For instance, benzophenone-3 (0.2-13 ng g), bisphenol A (7.7-13.7 ng g), methyl 3,5-dihydroxybenzoate (8-254 ng g), mono butyl phthalate (2-17 ng g) and triclosan (0.3-9 ng g) were found at the highest concentrations, but the presence of other analogues was detected as well. The developed target method was further extended to suspect and non-target screening (SNTS) by means of LC coupled to high-resolution MS/MS. First, well-defined workflows for SNTS were validated by applying the previously developed method to an extended list of compounds (83), and then, to the same real urine samples. From a list of approximately 4000 suspects, 33 were annotated at levels from 1 to 3, with food additives/ingredients and personal care products being the most abundant ones. In the non-target approach, the search was limited to molecules containing S, Cl and/or Br atoms, annotating 4 pharmaceuticals. The results from this study showed that the combination of the lower limits of detection of MS/MS and the identification power of high-resolution MS/MS is still compulsory for a more accurate definition of human exposome in urine samples.
本工作开发了一种同时测定尿液中 24 种不同内分泌干扰化合物（EDCs）的目标分析方法（包括酶解、固相萃取净化和液相色谱-串联质谱联用分析）。对目标分析方法（包括酶解、固相萃取净化和液相色谱-串联质谱联用分析）进行了优化、验证，并应用于志愿者样本，其中 67%的目标 EDCs 被定量。例如，在志愿者样本中检测到最高浓度的二苯甲酮-3（0.2-13ng g）、双酚 A（7.7-13.7ng g）、甲基 3,5-二羟基苯甲酸酯（8-254ng g）、邻苯二甲酸单丁酯（2-17ng g）和三氯生（0.3-9ng g），但也检测到其他类似物的存在。该方法进一步通过液相色谱-高分辨质谱联用进行了疑似和非目标筛查（SNTS）。首先，通过将之前开发的方法应用于扩展的化合物列表（83 种），并进一步应用于相同的真实尿液样本，对 SNTS 的定义明确的工作流程进行了验证。从大约 4000 个可疑物列表中，有 33 个物质被注释到了 1-3 级，其中食品添加剂/成分和个人护理产品是最丰富的。在非目标方法中，搜索仅限于含有 S、Cl 和/或 Br 原子的分子，注释了 4 种药物。本研究结果表明，对于更准确地定义尿液样本中的人类暴露组，仍然需要将 MS/MS 的检测下限和高分辨 MS/MS 的鉴定能力相结合。
