Application of novel analytical techniques and data analysis tools for comprehensive metabolomics analysis of complex biological mixtures
Vladimir Shulaev
University of North Texas, USA
: J Diagnos Tech Biomed Anal
Abstract
Metabolomics analysis of complex mixtures using a single separation technique is challenging due to the diversity of metabolites polarity, volatility and the large range of concentrations in biological samples. We examined two novel analytical approaches for comprehensive analysis of metabolites in various biological matrices. First approach is based on Ultra Performance Convergence Chromatography (UPC2), a chromatographic system that utilizes liquid CO2 as primary solvent to leverage the chromatographic principles and selectivity of normal phase chromatography while providing the ease-of-use of reversed-phase LC. We utilized the sub-2 µm particle supercritical chromatography for the separation of free fatty acids, neutral and polar lipids in a single lipid extract. Second approach utilizes Atmospheric pressure GC (APGC) is a ‘soft‘ chemical ionization technique that generates a spectrum conserving the molecular ion species with minimal fragmentation, which differentiates it from traditional vacuum source GC-MS based on electron ionization. We combined APGC with Ion Mobility (IM) to enhance peak capacity and improve selectivity and specificity of analysis. We demonstrated the utility of APGC-TOF-MS for metabolomics analysis of various mutant plant genotypes. Raw data were analyzed using TransOmics software that adopts an intuitive workflow approach to performing comparative metabolomics and lipidomics data analysis. The workflow starts with raw data file loading, then retention time alignment and deconvolution, followed by analysis that creates a list of features. The features are then identified with compound searches and explored using multivariate statistical methods.
Biography
Vladimir Shulaev holds dual PhDs in Biological Sciences and Plant Biology and has over 30 years research experience working at academia and industry. At UNT, he established the state of the art Metabolomics facility and heads the Center for Metabolomics and Metabolic Signaling Research and Biochemical Profiling Group. His research group develop novel analytical techniques for both targeted and non-targeted metabolomics using mass spectrometry platforms and applied metabolomics platform to systems biology, gene function elucidation, cancer development and progression, and modeling and simulation of biological networks.
