Specific determination of selenoaminoacids in whole milk by 2D size-exclusion-ion-paring reversed phase high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP MS) |
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Authors: | Bierla Katarzyna Szpunar Joanna Lobinski Ryszard |
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Institution: | a Laboratoire de Chimie Analytique Bio-Inorganique et Environnement, CNRS UMR5254, Hélioparc, 2, av. Pr. Angot, 64053 Pau, France b Warsaw Technical University, Department of Analytical Chemistry, Noakowskiego 3, 00-664 Warsaw, Poland |
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Abstract: | A procedure was developed for the quantitative recovery of selenomethionine (SeMet) and selenocysteine (SeCys) from whole milk. It was based on the protein unfolding, carbamidomethylation of the aminoacid residues using iodoacetamide and proteolysis using Protease XIV. The selenoaminoacids were specifically determined by ion-paring reversed phase HPLC-ICP MS after their isolation from the post-reaction mixture by size-exclusion LC. Se(IV) present in the sample was derivatized as well and was determined along with the selenoaminoacids. The origin and identity of species were identified by the co-elution with the Se(IV), isotopically labelled selenomethionine, and with the synthetic standard of carbamidomethylated selenocysteine. The method development for SeCys was assisted by using glutathione peroxidise as the SeCys standard. SeMet, SeCys and Se(IV) were quantified by the method of standard additions. The mass balance provided a measure of the method validation. The method was applied to monitoring selenium speciation during supplementation of cows (dose-effect study) with Se-rich yeast containing feed and during milk processing. |
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Keywords: | Milk Selenium Selenium speciation |
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