Abstract
Most analytical methods in metabolomics are based on one of two strategies. The first strategy is aimed at specifically analysing a limited number of known metabolites or compound classes. Alternatively, an unbiased approach can be used for profiling as many features as possible in a given metabolome without prior knowledge of the identity of these features. Using high-resolution mass spectrometry with instruments capable of measuring m/z ratios with sufficiently low mass measurement uncertainties and simultaneous high scan speeds, it is possible to combine these two strategies, allowing unbiased profiling of biological samples and targeted analysis of specific compounds at the same time without compromises. Such high mass accuracy and mass resolving power reduces the number of candidate metabolites occupying the same retention time and m/z ratio space to a minimum. In this study, we demonstrate how targeted analysis of phospholipids as well as unbiased profiling is achievable using a benchtop orbitrap instrument after high-speed reversed-phase chromatography. The ability to apply both strategies in one experiment is an important step forward in comprehensive analysis of the metabolome. Copyright © 2009 John Wiley & Sons, Ltd.
Highlights
Metabolomics and lipidomics are relatively new scientific disciplines, currently driven by the performance of the analytical instrumentation used
Highresolution orbitrap mass spectrometry is particular interesting for hyphenated LC/MS applications using ultraperformance liquid chromatography (UPLC) instruments, where sub-2 mm particle columns generate chromatography peaks with peak widths of only a few seconds, requiring at least 2 scans per second to obtain a sufficient number of data points across the peak for quantitation
The aim of the present study is to evaluate a secondgeneration, benchtop orbitrap mass spectrometry system for application to high-throughput metabolic profiling
Summary
Metabolomics and lipidomics are relatively new scientific disciplines, currently driven by the performance of the analytical instrumentation used. Some scientists argue that this method is of limited use when data will be integrated with other omics platforms such as proteomics or transcriptomics,[7] where unidentified analytes obstruct the formation and integration of biochemical networks For these researchers, it is useful to only collect data on known compounds as they can be interpreted in a biochemical sense.[8] As the specific, targeted analysis of known compounds demands a different method (e.g., tandem mass spectrometry, MS/MS) than global metabolic profiling (usually full scan MS spectra), the merger of the two. Ion chromatograms with a sufficiently high degree of accuracy, so that overlapping isobaric signals from salt adducts and lipids containing longer unsaturated fatty acids can be readily separated Such applications typically require 5 ppm or less mass measurement accuracy.[5]. We are describing the analysis of human plasma samples, with the goal of achieving simultaneous unbiased fingerprinting as well the targeted analysis of large numbers of metabolites within a single run, without compromising the analytical quality of the two strategies used
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