During COVID-19 outbreak, alcohol-based hand sanitizer (ABHS) has been widely used for hand cleaning and removing pathogens including human pathogens and zoonosis pathogens. High concentration of alcohol induces dehydrated skin in the users. Therefore, the objective of this study was to investigate moisturizing property and antimicrobial activity of alcohol-based hand sanitizer formulations using coconut oil as a moisturizing agent against pathogens including Escherichia coli and Staphylococcus aureus. The properties including antimicrobial activity, stability of the ABHS, and satisfaction levels of the coconut oil with two existing formulations by WHO and the Ministry of Public Health of Thailand (MOPH) were determined. The formulation containing the coconut oil demonstrated antibacterial activity against both E. coli and S. aureus with the minimal inhibitory concentration (MIC) of 8.75% V/V as same as MOPH. However, the WHO-modified formulation has the highest antimicrobial activity with the MIC value of 2.19% V/V. The stability result of 3 ABHSs showed that the preserved had the same efficacy compared to that of the fresh formulations. Hand washing with coconut formulation produced an average score of 7.19±1.71 on color, 7.06±1.56 on appearance, 5.82±2.10 on odor, 6.77±1.68 on moisture, 6.88±1.42 on overall acceptance out of 9. However, the sensory analysis of these three formulations results showed no significant difference in all parameters. The data suggest that moisturizing agents tested in our study do not affect the efficacy of ethanol. The coconut oil formulation is another good option for people searching for an effective hand sanitizer for germs protection and moisturizing.