Identification and quantification of the selenium species in biological tissues is imperative, considering the need to properly understand its metabolism and its importance in various field of sciences, especially nutrition science. Although a number of studies deals with the speciation of selenium, speciation analysis is still far from being a routine task, and so far strongly depends on the type of the samples. We present a study aimed to examine speciation analysis of Se in tissues of livers, muscles, and hearts obtained from lambs, namely in liver, muscle, and heart. The studied lambs were fed with the diet enriched with an inorganic (as sodium selenate) and organic chemical form of Se (as Se-enriched yeast) compounds with simultaneous addition of fish oil (FO) and carnosic acid (CA). The first part of the work was focused on the optimization of the extraction procedure of selenium compounds from tissues. Next, hyphenated high performance liquid chromatography and inductively coupled plasma mass spectrometry (HPLC-ICP-MS) was used for the identification of five seleno-compounds-Se-methionine (SeMet), Se-cystine (SeCys2), Se-methyl-Se-cysteine (SeMetSeCys), and Se(IV) and Se(VI). Verification of the identified seleno-compounds was achieved using triple-quadrupole mass spectrometer coupled to high performance liquid chromatography (HPLC-ESI-MS/MS). The applied procedure allowed for quantitative analysis of SeMet, SeCys2, and SeMetSeCys, in biological tissues. The developed analytical protocol is feasible for speciation analysis of small molecular seleno-compounds in animals samples.
Keywords: mass spectrometry; selenium species; selenium supplementation.