Growing antimicrobial resistance represents serious public health threat. With the increasing presence of online/e-pharmacies in India, public access to medicines has increased. Easy access coupled with lack of adequate and authentic drug information can undermine rational antibiotic use. The present study aimed at exploring the availability of antibiotics for online sale and comparing the e-pharmacies on key characteristics influencing safe and rational use of antibiotics. A cross-sectional study was conducted over a period of 3 months using the websites of 10 popular e-pharmacies in India. Availability of antibiotics for online sale was assessed in context of the National List of Essential Medicines, 2022 and WHO-AWaRe (Access, Watch, Reserve) categories. Three separate questionnaires were developed to extract and compare relevant information from e-pharmacies pertaining to their safety and authenticity (in light of the expected standards laid down in draft e-pharmacy rules, 2018), availability of drug product and consumer awareness information for promoting prudent antibiotic use. Data were summarized using descriptive statistics. Out of the 17 antibiotics studied; antibiotics belonging to all WHO-AWaRe categories i.e. Access (83.33%), Watch (88.33%) and Reserve (78%) groups were available through e-pharmacies. Wide variation exists among the e-pharmacies regarding compliance to studied parameters under three questionnaires. Increased access to Watch and Reserve groups of antibiotics can translate into antibiotic misuse. It is therefore crucial that the regulatory gaps concerning e-pharmacies are addressed urgently and consumers are educated regarding safe and rational use of antibiotics.