A next-generation sequencing approach for the detection of mixed species in canned tuna

•A next-generation sequencing method for canned tuna is presented.•The method detects different tuna species in mixtures.•Amplification of mtDNA markers cytb and control region.•Qualitative, and to some extent, semi-quantitative results.•Different bioinformatic analyses lead to similar results. Tuna cans are relevant seafood products for which mixtures of different tuna species are not allowed according to European regulations. In order to support the prevention of food fraud and mislabelling, a next-generation sequencing methodology based on mitochondrial cytochrome b and control region markers has been tested. Analyses of defined mixtures of DNA, fresh tissue and canned tissue revealed a qualitative and, to some extent, semiquantitative identification of tuna species. While the choice of the bioinformatic pipeline had no influence in the results (p = 0.71), quantitative differences occurred depending on the treatment of the sample, marker, species, and mixture (p