New analytical methods for studying low abundance metabolome

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The main objective of the emerging field of metabolomics is the analysis of all small molecule metabolites present in a particular biological system in order to provide better understanding of dynamic processes occurring in living systems. However, the analytical approaches for the comprehensive study of hundreds or thousands of metabolites in a single analysis are currently underdeveloped, better-suited for medium to high-abundance stable metabolites and lack adequate validation procedures. The objective of the proposed research program is to improve sample preparation methods in global metabolomics by liquid chromatography - mass spectrometry (LC-MS) with the primary focus of improving the detection and identification of low abundance metabolites without sacrificing high sample throughput and speed of analysis. In particular, we plan to examine the use of new extraction materials, nanoparticles and ionic liquids to identify the most useful new materials for the design of a sequential extraction/fractionation protocol based on magnetic separation platform. We will also devise new systematic protocol to evaluate the biological availability of metabolites in a given sample to determine both free and total metabolite concentrations, and evaluate whether this novel approach can improve the search for biomarkers. The research program will answer important questions not addressed in current metabolomics literature including (i) how to improve data quality while maintaining the global nature of analysis (ii) whether emerging materials such as customizable nanoparticles and ionic liquids can improve metabolite coverage over classical approaches (iii) how to identify the most complementary orthogonal methods for sample preparation and develop standardized sequential extraction protocols suitable for deep mining of the metabolome even in sample-limited situations and (iv) whether biological availability of metabolite can improve the processes of biomarker discovery and data interpretation. The new analytical methods developed in this program will play a critical role to improve both targeted and untargeted metabolomics studies, and facilitate faster progress and rational design of extraction methods in this new field.