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The plant defensive mechanisms against biotic and abiotic stresses involve the production of betaines and their structural analogs, the quaternary ammonium compounds (QACs). Numerous analytical methods have been employed for the detection and quantification of betaines in vegetable sources, including various types of chromatography and nuclear magnetic resonance. However, the breakthrough in the quantitative analysis of betaines has occurred with the advent of the mass spectrometric methodologies that allowed their specific quantification at very low concentration and with minimal sample manipulations. Indeed, betaines and QACs are characterized by the presence of a quaternary ammonium group that confers particular sensitivity and fragmentation features useful for their detection at low levels and selective quantification by mass spectrometry. This chapter focuses on the analytical methods for determination of betaines and QACs with particular attention to the high-performance liquid chromatography–electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS). Moreover, the quantitative distribution of betaines and QACs in fruits is also reported along with new knowledge about Nε-trimethyllysine, a less-known betaine recently found to be ubiquitous in the plant world.

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