Non-animal photosafety screening for complex cosmetic ingredients with photochemical and photobiochemical assessment tools

•New assay strategy was proposed for photosafety of complex cosmetic ingredients.•The maximum UV/VIS absorption value of 1.0 was adopted as a tentative criterion.•The ROS assay at 50μg/mL showed a high sensitivity to predict photoreactivity.•The 3T3 NRU PT would be useful as a follow-up assay to predict phototoxic risk. Previously, a non-animal screening approach was proposed for evaluating photosafety of cosmetic ingredients by means of in vitro photochemical and photobiochemical assays; however, complex cosmetic ingredients, such as plant extracts and polymers, could not be evaluated because their molecular weight is often poorly defined and so their molar concentration cannot be calculated. The aim of the present investigation was to establish a photosafety screen for complex cosmetic ingredients by using appropriately modified in vitro photosafety assays. Twenty plant extracts were selected as model materials on the basis of photosafety information, and their phototoxic potentials were assessed by means of ultraviolet (UV)/visible light (VIS) spectral analysis, reactive oxygen species (ROS)/micellar ROS (mROS) assays, and 3T3 neutral red uptake phototoxicity testing (3T3 NRU PT). The maximum UV/VIS absorption value was employed as a judgment factor for evaluating photoexcitability of samples, and the value of 1.0 was adopted as a tentative criterion for photosafety identification. The ROS/mROS assays were conducted at 50μg/mL, and no false negative prediction was obtained. Furthermore, the ROS/mROS assays at 50μg/mL had a similar predictive capacity to the ROS/mROS assays in the previous study. A systematic tiered approach for simple and rapid non-animal photosafety evaluation of complex cosmetic ingredients can be constructed using these modified in vitro photochemical assays.