Abstract
A fit for purpose analytical protocol was designed towards searching for low molecular weight seleno-compounds in sprouts. Complementary analytical techniques were used to collect information enabling the characterization of selenium speciation. Conceiving the overall characterization of the behavior of selenium, inductively plasma optical mass spectrometry (ICP-MS) was used to determine the total selenium content in entire sprouts as well as in selected extracts or chromatographic fractions. Then, high-performance liquid chromatography combined with ICP-MS (HPLC-ICP-MS) was used to evaluate the presence of inorganic and organic seleno-compounds, with the advantages of being very sensitive towards selenium, but limited by available selenium standard compounds. Finally, ultra-high performance liquid chromatography electrospray ionization triple quadrupole mass spectrometry (UHPLC-ESI-QqQ-MS/MS) and UHPLC-ESI-Orbitrap-MS/MS were used for the confirmation of the identity of selected compounds and identification of several unknown compounds of selenium in vegetable sprouts (sunflower, onion, radish), respectively. Cultivation of plants was designed to supplement sprouts with selenium by using solutions of selenium (IV) at the concentration of 10, 20, 40, and 60 mg/L. The applied methodology allowed to justify that vegetable sprouts metabolize inorganic selenium to a number of organic derivatives, such as seleno-methylselenocysteine (SeMetSeCys), selenomethionine (SeMet), 5′-seleno-adenosine, 2,3-DHP-selenolanthionine, Se-S conjugate of cysteine-selenoglutathione, 2,3-DHP-selenocysteine-cysteine, 2,3-DHP-selenocysteine-cysteinealanine, glutathione-2,3-DHP-selenocysteine, gamma-Glu-MetSeCys or glutamyl-glycinyl-N-2,3-DHP-selenocysteine.
Highlights
Speciation studies are commonly conducted towards chemical characterization of various objects in respect to the chemical form of the element of interest [1]
We aim to evaluate the analytical performance of the developed approach combining complementary analytical techniques used to collect information enabling the broad characterization of selenium speciation in plants
The aim of the described project was to optimize the analytical methodology towards the determination of selenium as well as identify and characterize low-molecular-weight seleno-compounds in sprouts
Summary
Speciation studies are commonly conducted towards chemical characterization of various objects in respect to the chemical form of the element of interest [1]. The use of a combined analytical approach becomes a powerful tool towards studying new selenium compounds and their metabolic pathways in plants. Selenomethionine is the most studied organic selenium compound, it would be very valuable to identify other selenium compounds present in plants, which could be effective in cancer prevention. Our work aimed to validate the analytical protocol involving mass spectrometry techniques to study selenium biotransformation by sprouts and to apply this protocol to evaluate the total content of selenium as well as to identify known, and characterize unknown, selenium compounds in selected plants of nutrition capability. The characterization of selenium compounds found in sprouts was performed by ICP-MS, HPLC-ICP-MS, ultra-high performance liquid chromatography electrospray ionization QqQ mass spectrometry (UHPLC-ESI-QqQ-MS/MS), and UHPLC-ESI-Orbitrap-MS/MS
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