A simplified method for inorganic selenium and selenoaminoacids speciation based on HPLC-TR-HG-AFS

A simplified speciation method for the determination of selenite, selenate and three selenoaminoacids (selenocystine, selenomethylselenocysteine and selenomethionine) has being developed, based on the coupling of high performance liquid chromatography (HPLC), thermoreduction (TR), hydride generation (HG) and atomic fluorescence spectrometry (AFS). Most of the existing methods based on AFS detection employ a two step procedure to reduce selenate to selenite before HG: (i) Ultraviolet radiation followed by (ii) heating, to produce volatile hydrides of the selenium compounds. The proposed simplified method HPLC-TR-HG-AFS does not require ultraviolet radiation. Instead, KBr dissolved in a HCl solution is added during the heating step (thermoreduction), resulting in an effective hydride generation of the selenium species. Different variables (temperature, HCl and NaBH4 concentrations) have been optimized, using both univariant and multivariant experimental designs. The proposed method is therefore less complex and allows limits of detection, reproducibility and repeatability values similar or better than the existing AFS detection methods described in the literature. A Certified Reference Material (SELM-1 with certified selenomethionine content) and a Se-enriched algae sample have been successfully analyzed with the proposed method. The results were also compared to an alternative technique (GC-MS) that provided similar results.