In this study, we examined the distribution of polycyclic aromatic hydrocarbons (PAHs) in sediments from the Mahanadi River Estuary (MRE), identified sources, and evaluated the ecological toxicity. The PAHs distributions in MRE ranged from 13.1 to 685.4 ng g-1 (dry weight), with a mean value of 192.91 ± 177.56 ng g-1 (dry weight). Sediments at sites S11, S8, and S13 have the highest 3-rings, 4-rings, and 5-rings PAHs, respectively. In MRE, pyrene has a significantly higher concentration with a mean value of 30.51 ng g-1, followed by Fluoranthene (86.2 ng g-1), Chrysene (67.4 ng g-1), and Benzo(k)fluoranthene (54.2 ng g-1). Site S8 had a higher total PAH concentration than sites S11, S13, and S1. The diagnostic and principal component analysis suggests that PAHs originated from petroleum, oil, biomass, and coal combustion. Higher toxic and mutagenic equivalent quotients indicate potential aquatic toxicity and a need for continuous monitoring of MRE for PAHs pollution.