Introduction: Healthcare workers often experience skin dryness and irritation from performing hand hygiene frequently. Low acceptability and tolerability of a formulation are barriers to hand hygiene compliance, though little research has been conducted on what specific types of formulation have higher acceptability than others. Objective To compare the acceptability and tolerability of an ethanol-based ABHR gel with superfatting agents to the isopropanol-based formulations (a rub and a gel formulation) currently used by healthcare workers at the University of Geneva Hospitals, Geneva, Switzerland. Methods Forty-two participants were randomized to two sequences, testing the isopropanol-based formulation that they are using currently (Hopirub® or Hopigel®), and the ethanol-based formulation containing superfatting agents (Saniswiss Sanitizer Hands H1). Participants tested each of the formulations over 7–10 day work shifts, after which skin condition was assessed and feedback was collected. Results H1 scored significantly better than the control formulations for skin dryness (P = 0.0209), and participants felt less discomfort in their hands when using that formulation (P = 0.0448). H1 caused less skin dryness than Hopirub®/Hopigel® (P = 0.021. Though overall preference was quite polarized, 21 participants preferred H1 intervention formulation and 17 preferred the Hopirub®/Hopigel® formulation that they normally used in their care activities. Conclusion We observed a difference in acceptability and strongly polarized preferences among the participants' reactions to the formulations tested. These results indicate that giving healthcare workers a choice between different high-quality products is important to ensure maximum acceptability.