Confocal Raman mapping (CRM) is a powerful, label free, non‐destructive tool, enabling molecular characterization of human skin with applications in the dermo‐cosmetic field. Coupling CRM to multivariate analysis can be used to monitor the penetration and permeation of active cosmetic ingredients (ACI) after topical application. It is presently illustrated how multivariate curve resolution alternating least squares (MCR‐ALS) can be applied to detect and semi‐quantitatively describe the diffusion profile of Delipidol, a commercially available slimming ACI, from Raman spectral maps. Although the analysis outcome can be critically dependent on the a priori selection of the number of regression components, it is demonstrated that profiling of the kinetics of diffusion into the skin can be established with or without additionnal spectral equality constraints in the multivariate analysis, with similar results. Ultimately, MCR‐ALS, applied without spectral equality contraints, specifically identifies the ACI as one of main spectral components enabling to investigate its distribution and penetration into the stratum corneum and underlying epidermis layers.