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Diploma thesis deals with the experimental study of transdermal transport of individual hydroxyacids and their combination with water-soluble vitamins. During this work the literature review was prepared, which was focused on the use of hydroxyacids in the cosmetic and pharmaceutical industries. At the same time the current state of use of water-soluble vitamins for topical applications was studied. Series of experiments was performed using Franz diffusion cells to simulate transdermal transport in vitro across cutaneous, synthetic and artificial Strat M ® membranes. The methodology of sample preparation and analysis of their transport properties was invented and optimized. The optimized methodology was subsequently used in a series of diffusion experiments in which the penetrated amount of hydroxyacids was analyzed by ion chromatography. The concentration of water-soluble vitamins was determined by HPLC. All examined active substances demonstrate positive penetration properties. The best results were obtained with combination of glycolic acid with pyridoxine.