New Advances in Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry for Accelerating Biomarker Discovery in Metabolomics

代谢组学是一个多学科领域的发现为基础的研究，其目的是全面分析低分子量代谢物（通过开发基于多段注射毛细管电泳-质谱（MSI-CE-MS）技术的尖端方法克服这些障碍，该技术为代谢组学提供了一种具有高样品通量，严格质量控制和长期批次校正的变革性方法。我们的提案将大大扩大代谢组的覆盖范围，以实现快速，灵敏而稳健的分析，涵盖亲水性代谢物和完整的磷脂，包括具有生物学意义的低丰度代谢物，如D-氨基酸，生物胺和维生素D。这将通过开发相容的非水缓冲系统结合化学选择性衍生化策略来实现，同时使用新型纳米喷雾离子源来提高浓度灵敏度。由于质谱信息在分离时间内编码，因此将开发开放访问软件工具，以实现MSI-CE-MS数据的自动化处理，与传统的LC-MS，GC-MS和NMR平台相比，MSI-CE-MS数据在吞吐量，再现性和数据保真度方面无与伦比。此外，还将对不同类别的代谢物进行溶剂化离子电泳迁移率及其气相电离反应的基础研究，作为进行虚拟代谢组学研究的一种方式;这项工作将提供一种创新的策略，在缺乏化学标准品时，根据未知代谢物的推定化学结构与高分辨率MS/MS互补，质谱数据库仍然不完整。我们的研究计划还将使用最先进的MSI-CE-MS方法，应用于基于粪便代谢组表征的人类微生物组功能分析，以及识别与消防员烟雾暴露相关的新化学特征从体积限制的生物标本，包括使用非侵入性收集设备的汗液。这一及时的提案将提供开创性的分析技术，集成软件工具和加速的生物标志物发现数据工作流程，以支持全球代谢组学计划，该计划也具有强大的商业潜力，对加拿大的公共政策产生直接影响。

Metabolomics is a multidisciplinary area of discovery-based research whose aim is the comprehensive analysis of low molecular weight metabolites (overcome these obstacles by developing cutting-edge methodologies based on multisegment injection capillary electrophoresis-mass spectrometry (MSI-CE-MS) technology that offers a transformative approach to metabolomics with high sample throughput, stringent quality control and long-term batch-correction. Our proposal will greatly expand metabolome coverage to enable rapid, sensitive yet robust analyses spanning hydrophilic metabolites and intact phospholipids, including low abundance metabolites of biological significance, such as D-amino acids, biogenic amines, and vitamin D. This will be achieved by developing compatible nonaqueous buffer systems in conjunction with chemoselective derivatization strategies while using a novel nanospray ion source to boost concentration sensitivity. Since mass spectral information is encoded temporally within a separation, open-access software tools will be developed to enable automated processing of MSI-CE-MS data that is unsurpassed in throughput, reproducibility, and data fidelity as compared to conventional LC-MS, GC-MS and NMR platforms. Also, fundamental studies of the electrophoretic mobility of solvated ions and their gas-phase ionization responses will be investigated for diverse classes of metabolites as a way to conduct virtual metabololomic studies; this work will provide an innovative strategy to identify and quantify unknown metabolites based on their putative chemical structure complementary to high resolution MS/MS when chemical standards are lacking, and mass spectral databases remain incomplete. Our research program will also use state-of-the-art MSI-CE-MS methods as applied to the functional analysis of the human microbiome based on characterization of the fecal metabolome, as well as the identification of new chemical signatures associated with smoke exposure in firefighters from volume-restricted biospecimens, including sweat using a non-invasive collection device. This timely proposal will deliver groundbreaking analytical technology, integrative software tools, and accelerated data workflows for biomarker discovery in support of global metabolomic initiatives that also has strong commercial potential with direct impacts on public policy in Canada.