Introduction: Ensuring the safety of products causing ocular irritation is crucial. The Bovine Corneal Opacity and Permeability (BCOP) test is an accepted alternative method for ocular irritation assessment in regulatory contexts. Objective: This study aimed to identify the main challenges and technical limitations faced by professionals when implementing alternative methods, particularly for ocular irritation assessment. Method: A qualitative research approach was employed using questionnaires (n = 22 respondents) and interviews (n = 7), followed by discourse analysis. Results: Findings reveal that 71% of alternative method practitioners encounter significant difficulties, primarily due to high validation/implementation costs (62%), driven by consumable expenses. Limited access to training (62%) and importation of reagents and equipment (43%) were also noted. Key BCOP limitations include restricted access to bovine eyeballs (50%), less precise risk classification (43%), and substantial eyeball waste (29%) due to damage. Interviews supported these conclusions, highlighting challenges such as travel to slaughterhouses, insufficient technical details in the OECD Guide, the need for additional support materials, and a lack of experienced professionals. These results underscore the need for expanded training opportunities for test implementation. The high cost of imported materials indicates greater validation expenses in Brazil than in other countries. Conclusions: Addressing these challenges requires an open innovation environment, fostering collaboration among the Triple Helix (Companies, Scientific and Technological Institutions, and Government).