Make-up accessories may be subjected to bacterial contamination due to their contact with many surfaces, including the client's skin. Disinfection prevents the spread of infection, but the capacity to eliminate microorganisms varies among disinfection methods. Little is known about disinfection in beauty salons; therefore, we aimed to compare the effectiveness of disinfectants for reusable make-up applicators. We tested reusable make-up applicators (natural brush, synthetic brush, and sponge) with disinfectants (bactericidal soap, 70% ethanol, and Hydro Sept solution). The number of microorganisms present before and after disinfection using the plate method was assessed. In comparison to water, bactericidal soap reduced the number of bacterial colonies. There were 418 versus 83 colonies in natural brush samples, 1,162 versus nine colonies in synthetic brush samples, and 617 versus three colonies in sponge samples. Ethanol eliminated 100% of bacterial growth. Disinfection was more effective on applicators used for dry products (natural brush) than for wet products (sponge, synthetic brush). We conclude that disinfection is a complex process and depends on the type of make-up applicator being disinfected and the type of cosmetics being used. Further research on the selection of appropriate disinfection procedures for individual applicants and for those who prepare makeup is needed.