Glycerin is widely used in cosmetics and well as in pharmaceutical formulations, mainly as humectant. In vitro studies have shown glycerin to prevent crystallization of stratum corneum model lipid mixture at low room humidity. Whether this may affect the skin barrier function during repeated application of glycerin in a cream base to normal skin is not known. Therefore, the influence of a cream containing 20% glycerin was compared with its placebo cream in a bilateral, double-blind study on 17 healthy volunteers. The effect was evaluated as influence on hydration with a corneometer and on skin barrier function. Skin barrier function was assessed as permeability to water with an evaporimeter (transepidermal water loss; TEWL) and as sensitivity to an irritating surfactant by measuring the biological response (measured as TEWL and skin blood flow). Ten days treatment of normal skin with 20% glycerin significantly increased skin corneometer values, indicating an increased hydration. However, our study failed to show an influence of glycerin on human skin, in terms of TEWL and skin sensitivity to SLS-induced irritation.