There are various types of cytotoxic agents used in the treatment of chemotherapy, and each agent can cause undesirable side effects in both patients and nurses, who are also skilled in dealing with these antineoplastic agents. Twenty true or false, closed-ended questionnaires were utilised to assess and evaluate the knowledge of nurses regarding the safety of chemotherapy. A total of 107 nurses participated in the assessment and evaluation, with only 59.22% receiving correct answers overall. The majority of respondents, 73.8% (79 out of 107) demonstrated moderate knowledge with 10 to 16 correct responses out of 20 questions. The majority of their knowledge regarding the safe handling of chemotherapy came from Continuous Nursing/Medical Education (CNE/CME) and the internet respectively. In general, the results demonstrated that nurses have insufficient knowledge of cytotoxic agents, as evidenced by the fact that the average percentage of correct responses was below 60%. Therefore, nurses must receive extensive additional education about anticancer agents in nursing school and through in-hospital continuing education. It is essential for nurses to participate in a structured educational programme, such as a Post Basic or Advanced Diploma in Oncology Care, in order to obtain proper updates and certification. In addition, the study suggested that updating the knowledge of safe handling among nurses with supported guidelines and policies will aid in enforcing safety implementation within clinical settings.