Comprehensive Analytical Strategy for Biomarker Identification based on Liquid Chromatography Coupled to Mass Spectrometry and New Candidate Confirmation Tools

Citations Over TimeTop 11% of 2009 papers

Abstract

A comprehensive analytical LC-MS(/MS) platform for low weight biomarkers molecule in biological fluids is described. Two complementary retention mechanisms were used in HPLC by optimizing the chromatographic conditions for a reversed-phase column and a hydrophilic interaction chromatography column. LC separation was coupled to mass spectrometry by using an electrospray ionization operating in positive polarity mode. This strategy enables us to correctly retain and separate hydrophobic as well as polar analytes. For that purpose artificial model study samples were generated with a mixture of 38 well characterized compounds likely to be present in biofluids. The set of compounds was used as a standard aqueous mixture or was spiked into urine at different concentration levels to investigate the capability of the LC-MS(/MS) platform to detect variations across biological samples. Unsupervised data analysis by principal component analysis was performed and followed by principal component variable grouping to find correlated variables. This tool allows us to distinguish three main groups whose variables belong to (a) background ions (found in all type of samples), (b) ions distinguishing urine samples from aqueous standard and blank samples, (c) ions related to the spiked compounds. Interpretation of these groups allows us to identify and eliminate isotopes, adducts, fragments, etc. and to generate a reduced list of m/z candidates. This list is then submitted to the prototype MZSearcher software tool which simultaneously searches several lists of potential metabolites extracted from metabolomics databases (e.g., KEGG, HMDB, etc) to propose biomarker candidates. Structural confirmation of these candidates was done off-line by fraction collection followed by nanoelectrospray infusion to provide high quality MS/MS data for spectral database queries.

Related Papers

• → Analytical Characterization of the Electrospray Ion Source in the Nanoflow Regime(2008)90 cited
• → Suppression of Protein Structural Perturbations in Native Electrospray Ionization during the Final Evaporation Stages Revealed by Molecular Dynamics Simulations(2021)11 cited
• → Electro-kinetic assisted electrospray ionization for enhanced complex sample analysis(2016)10 cited
• → Pneumatically assisted electrospray-ion mobility spectrometry for quantitative analysis of intact proteins(2012)7 cited
• → Chiral recognition and enantiomer assays of N-(3,5-dinitrobenzoyl)amino acid derivatives using electrospray ionization–mass spectrometry(2009)4 cited