Humans are at risk of exogenous exposure to exogenous chemicals. Challenges exist for the comprehensive monitoring of residues with different physical and chemical properties in serum. Here, an on-line two-dimensional liquid chromatography (2D-LC) - high resolution mass spectrometry system (HRMS) was developed, expanding the range of the partition coefficient in octanol/water of the residue analysis from −8 to 12. A high-coverage serum residue screening strategy was further designed by integrating 2D-LC system with HRMS full MS/data independent acquisition and automatic spectral library searching. This strategy enables to simultaneously screen 1210 pesticides, veterinary/human drugs, other chemical pollutants and their metabolites in serum with a single analysis. Method validation showed 92% and 81% of 1022 residues spiked in serum could be detected at 50 ng/mL and 5 ng/mL, respectively. The developed method was applied to the analysis of 24 separately pooled serum samples, 58 suspect residues were found, some of them were detected at high frequencies over than 50%. Among them, 4,6-Dinitro-O-cresol and probable carcinogenic folpet are highly toxic, and cimaterol is banned in China. Collectively, this study developed a 2D-LC-HRMS -based screening strategy for screening pesticides, veterinary/human drugs, and other chemical pollutants in serum, it is helpful for studying the effect of exogenous exposures on human health.

人类有暴露于外源性化学物质的风险。血清中不同理化性质的残留物的综合监测存在挑战。在此，开发了在线二维液相色谱 (2D-LC) - 高分辨率质谱系统 (HRMS)，将残留物分析的辛醇/水分配系数范围从 -8 扩大到 12。通过将 2D-LC 系统与 HRMS 全 MS/数据独立采集和自动谱库搜索相结合，进一步设计了高覆盖度血清残留筛选策略。该策略能够同时筛选 1210 种农药、兽药/人药、其他血清中化学污染物及其代谢物的单次分析。方法验证表明，在 50 ng/mL 和 5 ng/mL 的情况下，血清中加标的 1022 种残留物中的 92% 和 81% 分别可以被检测到。将开发的方法应用于 24 个单独合并的血清样品的分析，发现了 58 个可疑残留物，其中一些残留物的检测频率超过 50%。其中，4,6-二硝基-邻甲酚和可能致癌的灭菌菊剧毒，西马特罗在我国禁用。总的来说，本研究开发了一种基于 2D-LC-HRMS 的筛查策略，用于筛查血清中的农药、兽药/人药和其他化学污染物，有助于研究外源性暴露对人体健康的影响。