Four chimpanzee subspecies (Mammalia Hominidae) are commonly recognised: the Western Chimpanzee, P. troglodytes verus (Schwarz, 1934), the Nigeria-Cameroon Chimpanzee, P. troglodytes ellioti, the Central Chimpanzee, P. troglodytes troglodytes (Blumenbach, 1799), and the Eastern Chimpanzee, P. troglodytes schweinfurthii (Giglioli, 1872). Recent studies on mitochondrial DNA show the incorporation of P. troglodytes schweinfurthii in P. troglodytes troglodytes, suggesting the existence of only two subspecies: P. troglodytes troglodytes in Central and Eastern Africa and P. troglodytes verus-P. troglodytes ellioti in West Africa. The aim of the present study is twofold: first, to identify the correct subspecies of two chimpanzee samples collected in a Biopark structure in Carini (Sicily, Italy), and second, to verify whether there was a kinship relationship between the two samples through techniques such as DNA barcoding and microsatellite analysis. DNA was extracted from apes' buccal swabs, the cytochrome oxidase subunit 1 (COI) gene was amplified using universal primers, then purified and injected into capillary electrophoresis Genetic Analyzer ABI 3130 for sequencing. The sequence was searched on the NCBI Blast database. In addiction, the microsatellite analysis was performed on the same machine for parentage detection among samples, and data were analyzed with GenMapper software. Our results show that both samples were P. troglodytes troglodytes, while the analysis of the microsatellite results in an unclear relationship between two chimpanzee samples.