A New Derivatization Reagent for HPLC–MS Analysis of Biological Organic Acids
2017
Marquis, Bryce J. | Louks, Hayley P. | Bose, Chhanda | Wolfe, Robert R. | Singh, Sharda P.
Small molecules containing carboxylic acid functional groups are ubiquitous throughout biology, playing vital roles in biological chemistry ranging from energy metabolism to cellular signaling. This paper describes a new derivatization reagent, 4-bromo-N-methylbenzylamine, which was selected for its potential to derivatize mono-, di- and tri-carboxylic acids, such as the intermediates of the tricarboxylic acid (TCA) cycle. This derivatization procedure facilitated the use of positive electrospray ionization (ESI) tandem mass spectrometry (MS/MS) detection of derivatized species allowing for clear identification thanks to the easily recognizable isotope pattern of the incorporated bromine. A liquid chromatography (LC)-MS/MS method was developed which provided limits of detection between 0.2 and 44 μg L⁻¹ in under 6 min, depending on the analyte and total analysis time. This method was successfully applied in both in vitro and in vivo models.
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