Background: DNA barcoding depend on a piece of the cytochrome C oxidase subunit I (COI) gene in the mitochondrial genome is broadly useful in species ID and biodiversity studies. The aim of this study is to create a complete barcoding reference database of some fishes in the Mediterranean Sea fall under the family Sparidae. . Materials and methods: Mitochondrial COI barcode sequences were demonstrated from 8 species of family Sparidae in the order Perciformes, the mean length of Mitochondrial COI sequences was 650 base pairs. Results: The results of the phylogenetic tree presented that monophyly of Sparidae species. The studied species displayed clades of conspecific sequences and showed a match between the present study and the GenBank (NCBI) database. All groups clustered with high bootstrap value, that showed next to each branch and the tree was rooted based on the out group of Rhincodon typus. Conclusion: We achieve that COI sequencing can be used to recognize different fish species, and also it is used to obtain high competence of species reorganization by DNA barcoding. We underline the power of DNA barcode and its tools to identify different species from Mediterranean Sea. Results give Species ID for each species under study by using DNA barcoding.