While there is at least one standardized test available for evaluating the antioxidant capacity of cosmetic formulations, the authors proposed a new in vivo method to determine kinetics of squalene (SQ) photo-oxidation products (squalene monohydroperoxide, SQmOOH) as a reliable method with which to evaluate antioxidant capacity of a cosmetic formulation. Topical antioxidant formulation was applied on the foreheads of 30 volunteers. Sebum samples were collected before application of topical antioxidant and after one hour, three hours and five hours from the application of topical antioxidant. One half of the sebutape was irradiated and SQmOOH/SQ ratios in the skin lipid were analyzed using HPLC method. These results showed protective action of the topical antioxidant formulation against skin lipid peroxidation with a significant reduction of the quantity of SQmOOH (P<0.0001). Determination of the kinetics of squalene peroxidation is a reliable in vivo method in the evaluation of antioxidant capacity of cosmetic formulations.