Grant: Discovery Grants Program - Individual

Advances in mass spectrometry (MS) instrumentation now make it possible to identify thousands of biomolecules from a single biological sample. Despite this, in most experiments only a subset of the expected number of biomolecules are identified and reported in the literature. More recently, MS has emerged as a quantitative tool to monitor the dynamics of biomolecules during the progression of biological events or in response to external stimuli. A number of MS-based quantitative tools have been developed with which to study biomolecules, however few work on different biomolecule classes at the same time. Most are cost prohibitive and many cause the analytical sensitivity to decrease. There is a need for new developments in MS-based quantitative biomolecule analysis and this will continue to be the focus of the research program in the Smith laboratory. Efforts will be directed toward new innovations using Trimethylation Enhancement using Diazomethane (TrEnDi). TrEnDi is a chemical derivatization technique we developed that uses diazomethane to methylate functional groups that have pKa values lower than 11. In the case of peptides, lipids and metabolites, this translates into the neutralization of negatively charged functional groups and the formation of fixed and permanent positive charges to enhance ionization and increase sensitivity. Our research proposes to develop an entirely new strategy to conduct TrEnDi where diazomethane is created and used in situ. This development will enhance the safety of TrEnDi and enable different types of analytes to benefit from this powerful derivatization chemistry. The creation of an automated platform to conduct in situ TrEnDi (iTrEnDi) will allow greater control over the chemistry and enable high throughput analyses. Novel methods employing iTrEnDi to identify phosphopeptides, methylated peptides and glycerophospholipids at enhanced levels of sensitivity will be developed opening the door to greater insight into cellular biochemistry than possible before. For the first time, iTrEnDi will also be used to chemically derivatize small molecules found in biological fluids that originate from outside the body. Our efforts will focus on enhancing the detectability of fentanyl and carfentanil, two deadly opioids that are primarily responsible for the opioid crisis that we are facing in Canada. Our efforts will improve forensic and medical analyses by detecting these drugs at even lower concentrations. Finally, we will use iTrEnDi to enhance the analytical characteristics of glyphosate, the active ingredient in the controversial weed killer Roundup. We expect that our developments will enable glyphosate and other herbicides to be detected at lower concentrations than previously possible enabling rigorous environmental and biomedical studies. HQP will drive the research program in a highly collaborative and inclusive environment gaining skill sets that are attractive to the Canadian job market.