Purpose: Prevalence of skin cancer is rapidly increasing. There is a need for non-invasive biomarkers to assess efficacy of prevention strategies aiming at reduction of exposure to ultraviolet radiation (UVR). Recently, stratum corneum (SC) biomarkers were applied in various inflammatory skin diseases. Here, we explore their suitability as candidate biomarkers for UVR. Material and methods: Twelve volunteers were exposed to a UVB-dose of 0.72 SED, three times a week, during three weeks. As candidate biomarkers, cis-isomers of urocanic acid (cUCA) and 25 immunological mediators were measured in the SC. Results: Eight immunological markers significantly changed from baseline. Of them, IL-1RA/IL-1α and a placental growth factor (PIGF) showed gradual changes during UVR-exposure (p < 0.01 for linear trend). cUCA increased sharply already after the first exposure, however, reached a plateau in the second week. Conclusions: SC represents a promising, non-invasive alternative to skin biopsy in detecting UVR-induced changes. cUCA is the marker of choice for assessment of single UVR-exposure; however, it is less suitable for cumulative UVR-dose. Immunological markers including IL-1RA/IL-1α and PIGF showed gradual changes, and therefore are convenient for monitoring chronic UVR-exposure. These candidate biomarkers might facilitate assessment of the efficacy of preventive measures in the workplace and general population.