A study of low level selenium determination by hydride generation atomic fluorescence spectrometry in water soluble protein and peptide fractions

Development of a method for very low level selenium determination in water soluble protein and peptide fractions, obtained after various separation procedures, is presented. A hydride generation atomic fluorescence spectrometry (HG-AFS) detection system was optimised and the influence of Cu(II), Sb(...

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Bibliographic Details
Published in:Analytical and bioanalytical chemistry 2003-12, Vol.377 (7-8), p.1175-1183
Main Authors: Stibilj, V, Mazej, D, Falnoga, I
Format: Article
Language:English
Subjects:
Animals
Cell Fractionation
Chickens
Dietary Fats - analysis
Female
Liver - chemistry
Metallothionein - analysis
Models, Chemical
Peptides - chemistry
Proteins - chemistry
Selenium - analysis
Selenomethionine - analysis
Solubility
Spectrometry, Fluorescence - methods
Spectrophotometry, Atomic - methods
Water - chemistry
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Summary:Development of a method for very low level selenium determination in water soluble protein and peptide fractions, obtained after various separation procedures, is presented. A hydride generation atomic fluorescence spectrometry (HG-AFS) detection system was optimised and the influence of Cu(II), Sb(V), As(III) and HNO3 interferences in the measurement of Se by HG-AFS was investigated. A destruction procedure using HNO3 and H2O2 was also optimised and the average recovery of the digestion of a solution of selenomethioneine was 92 +/- 4% (n=14). Combination of this digestion with the detection system gave reliable results. Accuracy was tested by comparison with two independent methods. A very low detection limit (DL) of 0.2 ng/g of measuring solution was achieved. The whole procedure from weighing to measuring was performed in the same Teflon tube. The addition of HNO3 to the fractions before long term storage at -20 degrees C was necessary to prevent adsorption on the test tubes. Selenium was measured in water soluble protein and peptide fractions obtained after extraction, and Sephadex G-75 chromatography performed on liver samples from: i) hens exposed to As2O3, ii) hens fed with a high fat feed and iii) the certified reference material dogfish liver (CRM DOLT-2). Because of the very low DL we were able to observe the Se distribution in chromatographic fractions of samples of organisms which were not exposed to excess amounts of Se. The presence of selenium associated with metallothioneins was observed.
ISSN:1618-2642
1618-2650
DOI:10.1007/s00216-003-2182-9