RT Journal Article
T1 Simultaneous speciation of selenoproteins and selenometabolites in plasma and serum by dual size exclusion-affinity chromatography with online isotope dilution inductively coupled plasma mass spectrometry
A1 García Sevillano, Miguel Ángel
A1 García Barrera, Tamara
A1 Gómez Ariza, José Luis
AB A method for the simultaneous speciation of selenoproteins and selenometabolites in mouse plasma has been developed based on in series two-dimensional size exclusion and affinity high-performance liquid chromatography (2D/SE-AF-HPLC), using two columns of each type, and hyphenation to inductively coupled plasma-(quadrupole) mass spectrometry (ICP-QMS). The method allows the quantitative determination of selenoprotein P (SeP), extracellular glutathione peroxidase (eGPx), selenoalbumin (SeAlb), and selenometabolites in mouse plasma using species-unspecific isotope dilution (SUID). The 2D chromatographic separation is proposed to remove typical spectral interferences in plasma from chloride and bromide on 77Se (40Ar37Cl) and 82Se (81Br1H). In addition, the approach increases chromatographic resolution allowing the separation of eGPx from Se metabolites of low molecular mass. The method is robust, reliable, and fast with a typical chromatographic runtime less than 20 min. Precision in terms of relative standard deviation (n = 5) is in the order of 4 %, and detection limits are in the range of 0.2 to 1.0 ng Se g−1. Method accuracy for determination of total protein bound to Se was assessed by analyzing human serum reference material (BCR-637) certified for total Se content, and latterly applied to mouse plasma (Mus musculus). In summary, a reliable speciation method for the analysis of eGPx, selenometabolites, SeP, and SeAlb in plasma/serum samples is proposed for the first time and is applicable to the evaluation of Se status in human in clinical studies and other mammals for environmental or toxicological assessment.
PB Springer Verlag
SN 1618-2650
YR 2014
FD 2014
LK http://hdl.handle.net/10272/14475
UL http://hdl.handle.net/10272/14475
LA eng
NO García Sevillano, M.A., García Barrera, T., Gómez Ariza, J.L.: "Simultaneous speciation of selenoproteins and selenometabolites in plasma and serum by dual size exclusion-affinity chromatography with online isotope dilution inductively coupled plasma mass spectrometry".  	Analytical and Bioanalytical Chemistry. Vol. 406, n. 11, págs. 2719–2725, (2014). DOI: 10.1007/s00216-014-7629-7
NO This work was supported by the projects CTM2012-38720-C03-01 from Ministerio de Economia y Competitividad (Spain), and P008 FQM-3554 and P009-FQM-4659 from Consejeria de Innovacion, Ciencia y Empresa (Junta de Andalucia-Spain). Miguel Angel Garcia Sevillano thanks the Ministerio de Educacion for a predoctoral scholarship. We want to thank to the group of Prof. Sanz-Medel at the University of Oviedo, especially to Dr. Fernandez-Sanchez, for introducing and training in isotope dilution analysis.
DS Repositorio Institucional de la Universidad de Huelva
RD 1 jun 2026