Objectives: The aim of this research was to determine changes of the skin caused by long-term use of antiseptics and to investigate the impact of emollient preparations on skin treated with antiseptic by objective measurements of skin parameters. Subjects and methods: This research was conducted in 2022 at the University of Split School of Medicine in Croatia and it included 12 healthy subjects (N=12). In this randomized controlled trial, using Finn's chambers, antiseptic was applied to the subjects' volar side of the forearm during three weeks. Changes in skin parameters were monitored and compared to those of intact skin. In addition to that, the subjects used an emollient preparation on only one forearm in order to examine its effect on parameters of skin treated with antiseptic. Skin changes were monitored by measuring transepidermal water loss, skin hydration and erythema with MPA6 system probes (Courage+Khazaka, Germany). Results: Statistically significant changes were found in the values of transepidermal water loss and skin hydration after the exposure to an antiseptic compared to intact skin. However, there was no significant change in the values of erythema after the application of the antiseptic. The use of emollient preparation had a significant effect on skin hydration, but no significant changes were found in values of TEWL and erythema. Conclusions: Repeated application of antiseptic led to an increase in TEWL and a decrease in the skin hydration values, which indicates that it caused damage to the skin. The positive influence of the emollient preparation on skin hydration points to quicker healing of the skin barrier damaged by antiseptic.