Abstract
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.
Highlights
Aquaculture is one of the fastest-growing industries in animal food production, due to its health benefits and as a good source of proteins for humans (Ayadi, Rosentrater & Muthukumarappan, 2012)
The presented modified DNPH-based method showed to be suitable for the determination of protein oxidation of aquaculture feed samples, which are composed of manly of 24 kDa, 29 kDa and 66 kDa protein fractions
The results indicated that a long time of protein extraction, higher trichlorоacetic acid (TCA) concentration for protein precipitation, and higher ionic strength of extraction solution (0.5 M KCl) resulted in the highest protein content and the highest amount of carbonyls (7.96 nmol/mg protein) in the aqua feed sample
Summary
Aquaculture is one of the fastest-growing industries in animal food production, due to its health benefits and as a good source of proteins for humans (Ayadi, Rosentrater & Muthukumarappan, 2012). The global production of aquaculture feed increased by 4% in 2018 with an estimated 40 million tons, and this industry accounts for more than a. Dragojlović et al, Determination of protein oxidation in aquaculture feed, Food and Feed Research, 48 (2), 175-184 half of world fisheries production for human consumption (FAO, 2012). Contemporary manufactured aquaculture feed is prepared according to nutritional requirements of type and life stage of fish species (Davis, 2015). Oxidation of proteins is still not well investigated in the food and feed chain, and most previous studies have been conducted in order to investigate the involvement of protein oxidation in several human diseases (Davis, 2015; Reeg & Grune, 2015)
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