In Indonesia, the surge in online food Small and Medium Enterprises (SMEs) has underscored the critical need for stringent sanitation and hygiene practices. Utilizing a cross-sectional approach, the research assessed knowledge levels through a questionnaire, complemented by an evaluative tool aligned with guidelines from the Indonesian food and drug regulatory authority. From a sample of 132 SMEs spanning various Indonesian cities and regencies, it was observed that 57.6% had an adequate grasp of sanitation principles. Furthermore, a significant correlation between knowledge and its practical application emerged. Notably, there was a discernible difference in practice scores between participants with limited knowledge compared to those with a deeper understanding, evidenced by a mean difference of 10.62±3.77. This highlights a substantial inconsistency in translating knowledge into practice. Despite a reasonable level of awareness, the practical execution of sanitation and hygiene standards remains uneven among these enterprises. This gap indicates a need for focused interventions to enhance the consistent application of sanitation practices in the Indonesian online food SME industry, ensuring better compliance with established hygiene standards.