The industry production of juice accumulates large quantities of by-products such as seeds. Passion fruit seeds represent a valuable source of fatty acids and bioactive compounds. As a potential raw material for food production, it is important to assess the biocompatibility of new products to confirm their ingestion is secure for the consumers. Thus, the objective of this work was to evaluate the toxicity of yellow passion fruit (Passiflora edulis f flavicarpa) seed oil obtained by different extraction methods: cold pressing (CPO), ultrasound (USO) with ethanol at 40°C, Soxhlet (SOXO) with hexane at 65°C, and Accelerated Solvent Extraction (ASEO) with hexane at 40°C. The extracted oils were tested regarding their cytotoxicity by the MTT assay (the 3-(4,5-dimethylthiazole-2-il)-2,5-diphenyltetrazolium bromide) for 24h using HCT116 (human colorectal carcinoma), HT29 (human colorectal adenocarcinoma), HeLa (human cervical adenocarcinoma), and L-929 (mouse fibroblast) cell lines. The irritation potential by the Hen's Egg Test Chorioallantoic Membrane (HET-CAM). As results, cytotoxicity was not observed for all cell lines tested at maximum oil concentration of 100 mg/mL. The irritation potential test did not register inflammatory nor vascular effects (vasoconstriction, coagulation, and hemorrhage), once the scores corresponded to 0.0 (CPO, USO, and SOXO) and 0.72 (ASEO). Consequently, the oils evaluated can be classified as non-irritating substances, despite the application of different extraction methods. These results indicate that the purification of oils for solvent extraction was effective qualifying their application in food products. According to the tests performed, passion fruit seed oil is non-cytotoxic and non-irritating, therefore, considered biocompatible in the evaluated concentrations. seed oil represents a potential alternative for sustainable food production, representing a vegetable oil obtained through the conversion of agro-industrial waste into quality raw material, adding value to local agriculture and contributing to improve human health.