Original scientific paper

Uporaba voltmetrije za određivanje tiola i metalotioneina male molekularne mase u krvi svinje (Sus scrofa domestica)

Miroslava Beklova ; Department of Veterinary Ecology and Environmental Protection, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences, Palackeho 1-3, CZ-612 42 Brno, Czech Republic
Vojtech Adam ; Department of Chemistry and Biochemistry; Department of Animal Nutrition and Forage Production, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic
Petr Mares ; Department of Animal Nutrition and Forage Production, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic
Ladislav Zeman ; Department of Animal Nutrition and Forage Production, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic
Petr Dolezal ; Department of Animal Nutrition and Forage Production, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic
Jiri Pikula ; Department of Veterinary Ecology and Environmental Protection, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences, Palackeho 1-3, CZ-612 42 Brno, Czech Republic
Libuse Trnkova ; Department Chemistry, Faculty of Science, Masaryk University, Kotlarska 2, CZ-611 37 Brno
Rene Kizek ; Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic

Abstract

Metalotioneini (MT) igraju ključnu ulogu u održavanju homeostaze esencijalnih metala i u zaštiti od toksičnih metala te od oštećivanja stanice oksidacijom. Cilj ovog rada je predložiti novi pristup obradi bioloških uzoraka za analizu tiola, uključujući metalotioneine. Osim toga, predloženi postupak se testira pri kvantifikaciji MT i ukupnog sadržaja tiola u krvnom serumu svinje (Sus scrofa domestica), što se dosad nije radilo. Prikupljan je krvni serum (10 ml) te prebacivan u 0.2 M fosfatni pufer (990 ml). Uzorak (100 × razrijeđen) je stavljen u termomikser gdje se nastavila toplinska denaturacija većine proteina. Obrađeni uzorak seruma je elektrokemijski izmjeren da bi se odredio ukupni sadržaj tiola (cistein, glutation, metalotionein i drugi termostabilni tioli male molekularne mase) te sadržaj MT. Prosječna razina tiola i MT su procijenjeni na 165 ± 20 mM, odnosno 5.2 ± 0.6 mM.

Keywords

elektrokemijska metoda; voltmetrija diferencijalnim pulsom; Brdicka-reakcija; protein; svinja

Hrčak ID:

34030

URI

https://hrcak.srce.hr/34030

Publication date:

31.12.2008.

Article data in other languages: english

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