Natura Croatica Issue number: Natura Croatica, Vol. 17 No.4

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

Authors: Miroslava Beklova, Vojtech Adam, Petr Mares, Ladislav Zeman, Petr Dolezal, Jiri Pikula, Libuse Trnkova, Rene Kizek

Authors and affiliation

  • 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; 3Department 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 Republi

Abstract

Metallothioneins (MT) play a key role in maintaining the homeostasis of essential metals and in protecting of cells against metal toxicity as well as cell oxidative damaging. The aim of this work is to propose a new approach for processing a biological sample for analysis of thiols including metallothioneins. Moreover, the proposed procedure is tested on quantification of MT and total thiol content in blood serum of pig (Sus scrofa domestica), which has not been previously been performed. The blood serum (10 ml) was collected and transferred to 0.2 M phosphate buffer (990 ml). The sample (100 × diluted) was placed in a thermomixer, where heat denaturation of most of the proteins proceeded. The processed blood serum sample was electrochemically measured to determine total * Corresponding author: e-mail kizek@sci.muni.cz, phone: +420-5-4513-3350, fax: +420-5-4521-2044 Croatian Natural History Museum, Demetrova 1, Zagreb, Croatia 294 Beklova, M. et al.: Determination of low molecular mass thiols and metallothionein in blood of pig content of thiols (cysteine, glutathione, metallothionein and other low molecular thermostable thiols) and content of MT. The average level of the thiols and MT were estimated as 165 ± 20 mM and 5.2 ± 0.6 mM, respectively.

Keywords

electrochemical method, differential pulse voltammetry, Brdicka reaction, protein, pig