Background: Biomedical waste (BMW) generated in healthcare settings poses serious health and environmental risks if not managed properly. Nurses play a pivotal role in biomedical waste management; therefore, adequate knowledge among staff nurses is essential for effective implementation of biomedical waste management rules. Objective: To assess the knowledge regarding biomedical waste management among staff nurses working at Noor Hospital, Warudi, Jalna, and to determine the association between knowledge levels and selected demographic variables. Methods: A quantitative descriptive research design was adopted. Thirty staff nurses were selected using a non-probability purposive sampling technique. Data were collected using a structured knowledge questionnaire during January 2025. Descriptive and inferential statistics were used for data analysis. Results: The study revealed that 90% of staff nurses had average knowledge, 6.67% had good knowledge, and 3.33% had poor knowledge regarding biomedical waste management. A statistically significant association was found between knowledge levels and years of experience as well as designation (p < 0.05). Conclusion: Although most staff nurses demonstrated an average level of knowledge, gaps remain in comprehensive understanding of biomedical waste management. Regular training programs, continuous monitoring, and strict adherence to guidelines are recommended to improve compliance and ensure safe waste handling practices.