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Acid-base-driven matrix-assisted mass spectrometry for targeted metabolomics.

ABSTRACT: The ability to charge huge biomolecules without breaking them apart has made matrix-assisted laser desorption/ionization (MALDI) mass spectrometry an indispensable tool for biomolecular analysis. Conventional, empirically selected matrices produce abundant matrix ion clusters in the low-mass region (<500 Da), hampering the application of MALDI-MS to metabolomics. An ionization mode of MAILD, a rational protocol for matrix selection based on Brønsted-Lowry acid-base theory and its application to metabolomics, biological screening/profiling/imaging, and clinical diagnostics is illustrated. Numerous metabolites, covering important metabolic pathways (Krebs' cycle, fatty acid and glucosinolate biosynthesis), were detected in extracts, biofluids, and/or in biological tissues (Arabidopsis thaliana, Drosophila melanogaster, Acyrthosiphon pisum, and human blood). This approach moves matrix selection from "black art" to rational design and sets a paradigm for small-molecule analysis via MALDI-MS.

SUBMITTER: Shroff R

PROVIDER: S-EPMC2695405 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The lack of effective screening strategies for high-grade serous carcinoma (HGSC), a subtype of ovarian cancer (OC) responsible for 70-80% of OC related deaths, emphasizes the need for new diagnostic markers and a better understanding of disease pathogenesis. Capillary electrophoresis (CE) coupled with high-resolution mass spectrometry (HRMS) offers high selectivity and sensitivity for ionic compounds, thereby enhancing biomarker discovery. Recent advances in CE-MS include small, chip-based CE systems coupled with nanoelectrospray ionization (nanoESI) to provide rapid, high-resolution analysis of biological specimens. Here, we describe the development of a targeted microchip (µ) CE-HRMS method, with an acquisition time of only 3 min and sample injection volume of 4nL, to analyze 40 target metabolites in serum samples from a triple-mutant (TKO) mouse model of HGSC. Extracted ion electropherograms showed sharp, baseline resolved peak shapes, even for structural isomers such as leucine and isoleucine. All calibration curves of the analytes maintained good linearity with an average R2 of 0.994, while detection limits were in the nM range. Thirty metabolites were detected in mouse serum with recoveries ranging from 78 to 120%, indicating minimal ionization suppression and good accuracy. We applied the µCE-HRMS method to biweekly-collected serum samples from TKO and TKO control mice. A time-resolved analysis revealed characteristic temporal trends for amino acids, nucleosides, and amino acid derivatives. These metabolic alterations are indicative of altered nucleotide biosynthesis and amino acid metabolism in HGSC development and progression. A comparison of the µCE-HRMS dataset with non-targeted ultra-high performance liquid chromatography (UHPLC)-MS results showed identical temporal trends for the five metabolites detected with both platforms, indicating the µCE-HRMS method performed satisfactorily in terms of capturing metabolic reprogramming due to HGSC progression while reducing the total data collection time three-fold.

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Acid-base-driven matrix-assisted mass spectrometry for targeted metabolomics.

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