Background: Alcohol-based hand rub (ABHR) is widely used for hand disinfection in the health care sector. ABHR is, however, known to cause discomfort when applied on damaged skin emphasizing the unmet need for alternative and better tolerated types of disinfectants. Active chlorine hand disinfectants (ACHDs) are potential new candidates; however, the effect on the skin barrier function compared to ABHR remains to be assessed. Materials and methods: In Study A, the forearm skin of healthy adults was repeatedly exposed to ACHD and ABHR. Skin barrier function was assessed by measurement of transepidermal water loss, electrical conductance, pH, and erythema at baseline and at follow-up after 2 days, and subjective discomfort was likewise assessed. Study B was performed in the same way; however, in order to induce an experimental irritant contact dermatitis, sodium lauryl sulfate patch tests were applied to forearms before exposure to ACHD and ABHR. Results: In both studies, the skin barrier function was unaffected after repetitive exposure to ACHD and ABHR, and with no significant differences between the products. Subjective discomfort was reported as sporadic or very mild in relation to both products. Conclusion: Our results illustrate that use of ACHD does not affect the skin barrier function negatively, neither in intact skin nor in skin with experimentally induced contact dermatitis. Future studies should include real-life evaluation of skin barrier function and subjective discomfort following ACHD use in individuals with and without hand eczema.