Nitroaromatic compounds (NACs) are high of concern due to their mutagenicity, and carcinogenicity to organisms. Here, we attempted to establish a novel searching-validation-evaluation workflow that is tailored to recognize unknown NACs in environmental samples using liquid chromatography coupled with quadrupole Orbitrap high-resolution mass spectrometry (LC-Orbitrap-HRMS). We studied the fragmentation process of NAC standards in Orbitrap higher-energy collision dissociation (HCD) cells and observed that the mass loss of NO was the most prevalent among all NAC standards at both low and medium levels of collision energy. Thus, neutral loss of NO was considered as a diagnostic fragment of nitro groups and was used to screen out NACs in environmental samples. This technique is mass-loss-dependent, which enhances the recognition efficiency of NACs. Candidates exported from the PubChem compound database were further evaluated to obtain possible structures. This strategy was applied for the analysis of 24 surface soil, and we tentatively discovered two novel NACs in the analyzed samples. The semi-quantification results demonstrated that the concentrations of novel NACs were comparable to those of the ten targeted NACs in soil samples. This study provides an integrated strategy for the recognition of known and unknown NACs, which could be extended to other environmental matrices.

硝基芳族化合物 (NAC) 因其致突变性和对生物体的致癌性而备受关注。在这里，我们尝试建立一种新颖的搜索-验证-评估工作流程，该工作流程适用于使用液相色谱结合四极杆 Orbitrap 高分辨率质谱 (LC-Orbitrap-HRMS) 识别环境样品中的未知 NAC。我们研究了 Orbitrap 高能碰撞解离 (HCD) 电池中 NAC 标准的碎裂过程，并观察到在低和中等碰撞能量水平的所有 NAC 标准中，NO 的质量损失是最普遍的。因此，NO 的中性丢失被认为是硝基的诊断片段，并用于筛选环境样品中的 NAC。该技术依赖于质量损失，从而提高了 NAC 的识别效率。进一步评估从 PubChem 化合物数据库导出的候选物以获得可能的结构。该策略应用于 24 种表层土壤的分析，我们在分析的样品中初步发现了两种新型 NAC。半定量结果表明，新型 NACs 的浓度与土壤样品中十种目标 NACs 的浓度相当。该研究为识别已知和未知的 NAC 提供了一种综合策略，可以扩展到其他环境矩阵。半定量结果表明，新型 NACs 的浓度与土壤样品中十种目标 NACs 的浓度相当。该研究为识别已知和未知的 NAC 提供了一种综合策略，可以扩展到其他环境矩阵。半定量结果表明，新型 NACs 的浓度与土壤样品中十种目标 NACs 的浓度相当。该研究为识别已知和未知的 NAC 提供了一种综合策略，可以扩展到其他环境矩阵。