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

点击翻译按钮获取中文摘要

Metabolomics is a multidisciplinary area of discovery-based research whose aim is the comprehensive analysis of low molecular weight metabolites (< 1.0 kDa) in biological samples. Since metabolites are real-world endpoints of gene expression that also reflect environmental exposures, metabolomics offers a holistic approach to better understand the mechanisms underlying complex biological processes. However, there exists several "grand analytical challenges" that continues to hinder progress in global metabolomics initiatives, including low sample throughput, poor data fidelity, complicated data processing, limited metabolome coverage, and unknown compound identification. Our research program will 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.