Analytical and Bioanalytical Chemistry

, Volume 377, Issue 7–8, pp 1175–1183 | Cite as

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

  • V. StibiljEmail author
  • D. Mazej
  • I. Falnoga
Original Paper


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°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.


Selenium HG-AFS Liver Size exclusion chromatography Metallothioneins 



We would like to thank Drs. Zdenka Šlejkovec and Johannes van Elteren for their kind introduction to the HG-AFS technique. The Ministry of Education, Science and Sport of the Republic of Slovenia is thanked for financial support through the programmes P0–0531 "Biological and Geochemical Cycles" and P0–0532 "Radiochemistry and Radioecology".


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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  1. 1.Department of Environmental SciencesJožef Stefan Institute1000 LjubljanaSlovenia

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