干扰生物小分子代谢污染物的非靶向分析方法学研究

With the rapid growth of the number of commercial chemicals, high-throughout analysis of numerous contaminants and identification of toxic components in environment has become an analytical technology with urgent development. The project aimed to develop the non-targeted screening technologies for identification of compounds responsible for influencing biological metabolism. Based on the combination of new data-independent acquisition scanning mode, a full identification method will be established to obtain the corresponding MS/MS information of all signals with one injection. A highly sensitive method will be developed based on the QTOFMS NIS scanning mode together with chemical derivatizations, since some derivatization regents would specifically react with different functional groups and greatly enhance the detection sensitivities, resulting in the ultra-sensitive scanning of all the compounds containing certain functional groups. The above scanning technologies will be applied to biota samples to obtain the substance list in organisms. A high-throughout inquiry method for searching both intrinsic metabolites and exogenous pollutants was established. Then intrinsic biochemical chemicals will be distinguished from the exogenous pollutants based on the open databases. Base on the comparisons of the pollutants and metabolites, the toxic components will be identified with in vivo exposure experiments.

随着全球化学物质呈爆炸式增长，已知优控物质的环境暴露难以解释各种生态健康问题，如何从环境中高通量分析和甄别致毒污染物成为了重要的科学问题。本项目以非靶向分析的方法开发及鉴定干扰生物小分子代谢的污染物为目的开展研究，基于最新的数据非依赖型采集质谱技术，建立能够对所有物质两级质谱信号同时匹配的扫描鉴定方法，实现单次进样分析即获得全谱物质结构信息；利用衍生试剂与特定官能团的特异性反应及二级质谱分析的极高灵敏度，开发特异性高通量扫描的高灵敏分析方法；建立两级质谱信息与代谢组学谱库和污染物谱库的查询匹配方法，对生物内源性代谢分子和外源性污染物的进行区分扫描，并针对结构类似的污染物和关键代谢分子开展毒理研究，最终鉴定影响生物小分子代谢的环境污染物。

随着全球化学物质呈爆炸式增长，已知优控物质的环境暴露难以解释各种生态健康问题，如何从环境中高通量分析和甄别致毒污染物成为了重要的科学问题。该项目针对生物暴露的外源污染物及内源代谢小分子开展了高通量扫描技术、代谢行为及原因物质解析的研究工作。首先建立了基于数据非依赖型采集模式（DIA）的高分辨质谱两级信号匹配和鉴定的扫描方法，实现了单次进样获得扫描物质的全谱结构信息，并进一步建立了内源性代谢分子的代谢网络可视化方法，能够帮助快速识别出毒性污染物影响的关键代谢通路。发现了一系列化学衍生试剂与不同官能团的特异反应，显著增强质谱检测信号1-2个数量级，如：羧基物质与AMPP、氨基物质与SPTPP、及二氯甲烷对含氯物质氯增强电离；将衍生与高分辨质谱二级扫描有机结合，利用化学特异衍生与二级质谱扫描数据的计算技术，能同时对含目标官能团的上千种未知物质进行高通量扫描和鉴定。通过质谱扫描信息和不同谱库的高通量匹配比对，实现了生物内源性代谢分子和外源性污染物的区分扫描，发现了内源性代谢分子和外源性酚类污染物相互作用的成醚代谢新途径，并以太湖实际环境为调查区域，通过野生鲫鱼血液中内外源小分子的高通量鉴定，发现了不同区域鲫鱼血液中产生紊乱效应的代谢分子，并以此为靶点从鉴定的外源污染物中寻找原因物质。最终，结合区域差异、野生鲫鱼的物质暴露和组织分析，发现全氟十一酸的暴露是太湖野生鲫鱼甘油脂代谢紊乱的原因物质，建立了毒性污染物暴露和受干扰代谢分子的直接联系。该方法改变环境领域传统毒性物质鉴定方法和程序，提出了效应物质甄别的扫描技术和流程。