© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd Purpose: This study aimed to clarify the influence of different wiping methods on cleaning agent residue in hair follicles when using skin cleaning agents that are “removable-by-wiping.”. Methods: A total of 18 male volunteers were recruited and a cleaning agent containing 10% fluorescein sodium salt (cleaning agent) was used in this study. After gentle washing, the cleaning agent was removed via the washing method (control) or three distinct wiping methods (experiment). Hairs were obtained from the measurement area. The fluorescence intensities of the residual cleaning agent on the hair root and hair bulb were then evaluated after normalizing for autofluorescence from the hair. Fluorescence intensity was used to estimate the amount of cleaning agent residue. Results: No significant differences were found among cleaning agent removal methods (hair root: P = 0.67, hair bulb: P = 0.62) with respect to the amount of residual cleaning agent on hair. Conclusion: Cleaning agent residual index did not differ according to removal method. Cleaning agent remained in the hair follicle (hair roots, hair bulbs) despite removal from the skin by washing or wiping. Further studies are required to apply “removable-by-wiping” cleaning agents for people with vulnerable skin.