The 7th amendment of the EU Cosmetics Directive led to the ban of eye irritation testing for cosmeticingredients in animals, effective from March 11th 2009. Over the last 20 years, many efforts have beenmade to find reliable and relevant alternative methods. The SkinEthic™ HCE model was used to evaluatethe based on a specific 1-h treatment and a 16-h post-treatment incubation period was first assessed on a setof 102 substances. The prediction model (PM) based on a 50% viability cut-off, allowed to draw up twoclasses (Irritants and Non-Irritants), with good associated sensitivity (86.2%) and specificity (83.5%). Tocheck the robustness of the method, the evaluated set was expanded up to 435 substances. Final performancesmaintained a high level and were characterized by an overall accuracy value > 82% when usingEU or GHS classification rules. Results showed that the SkinEthic™ HCE test method is a promising in vitro eye irritancy potential of substances from a cosmetic industry portfolio. An optimized protocolin vitrotool for the prediction of eye irritancy. Optimization datasets were shared with the COLIPA Eye IrritationProject Team and ECVAM experts, and reviewed as part of an ongoing progression to enter an ECVAMprospective validation study for eye irritation.