Surfactants, which are widely used in skin care products and cleansers, can cause skin irritation. The skin irritation potential of surfactants is fundamentally determined by their molecular structure and is directly related to their microscopic aggregation structure and specific interactions with the skin. The microscopic origin of the irritation of the surfactants remains unknown. In this work, irritation properties of four surfactant solutions were measured, and their microscopic aggregation behavior was systematically analyzed. The results indicate that the surfactants self-assembled in aqueous solution to form aggregates with different morphologies, where the head groups of surfactants were closer to each other. Furthermore, surfactants that can form larger and more stable aggregate structures in aqueous solutions will exhibit less irritation. These findings hold significant implications for the design and expanded applications of mild surfactants.