Differentiated extraction strategies for selenium speciation in hyperaccumulator and non-hyperaccumulator plants via...

The growing demand for selenium (Se)-enriched products highlights the need for accurate and accessible Se speciation analysis. In this study, a cost-effective orthogonal liquid chromatography-atomic fluorescence spectrometry (LC-AFS) method was developed for the determination of seven Se species (SeO₃ ^2-, SeO₄ ^2-, SeLan, SeMet, SeCys₂, MeSeCys, and γ-GluMeSeCys), providing an alternative to ICP-MS-based approaches. Differences in Se accumulation mechanisms between hyperaccumulator Cardamine hupingshanensis and non-hyperaccumulator wheat significantly affected extraction efficiency. The sequential aqueous-enzymatic extraction increased SeLan recovery in C. hupingshanensis from 5.06 to 143.74 μg Se g^-1, a 28.41-fold increase, while anaerobic extraction enhanced SeMet recovery in wheat from 98.73 to 112.17 μg Se g^-1. Consequently, matrix-specific strategies improved Se recoveries from 20.82% to 80.65% in C. hupingshanensis and from 34.11% to 90.02% in wheat. These findings highlight the importance of matrix-specific extraction for accurate Se speciation analysis.