The critically endangered Brazilian guitarfish faces significant threats from environmental contamination. Assessing the impacts of such stressor is paramount from a conservational perspective. This study investigated the concentrations, distribution and accumulation patterns of organic contaminants in pregnant Brazilian guitarfish Pseudobatos horkelii. Blood, gill, gonad, liver, and muscle concentrations of polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), polybrominated diphenyl ethers, fecal sterols, and synthetic hormones used as human contraceptives were assessed. Synthetic hormones, especially D-norgestrel, showed the highest concentrations, mainly in the liver. Together with the results of fecal sterols, this finding suggests that guitarfish are exposed to sewage discharge. OCPs, especially hexachlorobenzene, mirex, endosulfans, and drins, showed considerably high concentrations, indicating the relevance of agricultural inputs. PCBs presented significant concentrations in the muscle, indicating long-term exposure, in contrast with other analytes that were primarily concentrated in the liver. These results have conservational implications, since contaminants analyzed herein have endocrine disruptive effects.