Determination of Protein Oxidation in Aquaculture Feed
The aim of this research was to develop reliable, easy-to-perform and cheap method for measuring protein oxidation in complex samples such as aquaculture feed within various protein sources. For that purpose modified 2,4-dinitrophenylhydrazyne (DNPH)-based method for quantification of protein carbonyls was employed, while the method modification was consisted of using different solutions for the extraction, time of protein extraction and concentration of trichloracetic acid (TCA) for protein precipitation. It was found that extraction during night, higher TCA concentration and the use of 0.5 M KCl extraction solution resulted in the highest protein amount measured by Lowry method and 280 nm protein estimation. On the other hand, the lowest protein yield was obtained by using distillated water for the extraction. Furthermore, the lowest amount of protein carbonyls was in the case when extraction was performed with distilled water (DW), while the highest content of protein carbonyls were reached with 0.15 M KCl and 0.5 M KCl extraction solutions. It was observed that the amount of carbonyls compounds were increasing during storage under accelerated conditions, and in comparison to the original (unmodified) method, the modified method for measuring protein oxidation resulted in the higher amount of carbonyls during the all points of storage.