Quantification of Se-Methylselenocysteine and Its γ-Glutamyl Derivative from Naturally Se-Enriched Green Bean (Phaseolus vulgaris vulgaris) After HPLC-ESI-TOF-MS and Orbitrap MS n -Based Identification

Orthogonal liquid chromatographic (ion exchange, reversed phase, and ion pairing) and mass spectrometric [electrospray ionization (ESI)-TOF-MS, ESI-Orbitrap MS, and inductively coupled plasma mass spectrometry (ICP-MS)] methods were addressed to identify and quantify selenium species from a naturally Se-enriched green bean (Phaseolus vulgaris vulgaris) sample after proteolytic digestion. While selenomethionine (10.1 mg/kg as Se) and selenate (9.5 mg/kg as Se) could be quantified in a straightforward way by anion exchange LC-ICP-MS technique, a multistep purification protocol was required to identify Se-methylselenocysteine and gamma-glutamyl-Se-methylselenocysteine in an unambiguous way prior to quantification by using either in-source fragmentation (LC-ESI-TOF-MS) or collision-induced dissociation (LC-ESI-Orbitrap MS). Finally, Se-methylselenocysteine (2.6 mg/kg as Se) and gamma-glutamyl-Se-methylselenocysteine (1.2 mg/kg as Se) could contribute to the overall selenium recovery of 72 %. This sample is the first of the Faboideae subfamily and Phaseolus ssp. to be speciated to such an extent for selenium including gamma-glutamyl-Se-methylselenocysteine, a highly potential selenium species, which makes this bean material an ideal candidate for functional food purposes.