Introduction: Formalin, which is widely used in anatomical laboratories exceeds the permissible range level. This has raised concerns about its effect on first professional medical students. Methods: The present study aims at assessing knowledge, attitude and practices of 200 first professional medical students about the use of formalin through a structured and validated questionnaire. Results: Most of the respondents answered satisfactorily about formalin and its composition. Irritation and burning sensation in eyes, nose, throat as well as lacrimation was proposed as the most common side effect of formalin (70%). Most of the respondents believed that existing infrastructure does not fulfill the required standards and formalin should be replaced by some other non–irritant chemical. Conclusion: The study identifies knowledge gaps and perceptions of medical students regarding use of formalin which will provide a breakthrough for replacing formalin with a better non–irritant chemical.