Comparative high-speed profiling of carboxylic acid metabolite levels by differential isotope-coded MALDI mass spectrometry.

This present work describes the development of a novel high throughput comparative matrix-assisted laser desorption ionization (MALDI) mass spectrometry profiling technique for endogenous compounds using a new isotope-coded label for relative quantitation of carboxylic acids. The key new aspect of this technique was a differential label, 3-hydroxymethyl-1-ethylpyrididinium iodide (HMEP), designed specifically for simultaneous quantitative MALDI analysis of two physiological states. The HMEP-d(0) and HMEP-d(5) variants of the label were applied to profiling endogenous fatty acid levels during a fish oil intervention study, using the metabolite profile of every individual person in the study as their own personal analytical reference standard. Initially, analytical figures of merit such as sensitivity, linear dynamic range, limit of quantitation, and precision were determined from the comparative quantitation experiments. Importantly, the permanently charged HMEP mass tag not only increased the ionization efficiency of the studied fatty acids but also ensured that the solution phase properties of the analytes became more similar, allowing the use of CHCA as a single MALDI matrix compound for the entire range of analytes. The label exhibited a further very unique feature; it provided complete suppression of MALDI matrix-related ions. The MALDI assay was able to generate the data much faster than conventional gas chromatography (GC) methods for fatty acids. It is shown in this study that analyzing a single sample took less than 10 s with analytical results of comparable quality to those obtained by GC.