Chemotherapeutic drugs improve cancer outcomes, but unsafe handling exposes nurses, patients, and healthcare environments to avoidable risks. This study assessed knowledge and practice of safe handling and administration of chemotherapeutic drugs among nurses in a tertiary health institution in Benin City, Edo State, Nigeria. A descriptive cross-sectional design was used, and all 278 registered nurses working in units involved in chemotherapy preparation, handling, transportation, storage, or administration were studied using a structured questionnaire; data were analysed with frequencies, percentages, mean values, and chi-square tests at a 0.05 significance level. Findings showed that most respondents had fair knowledge of chemotherapeutic drugs, while safe handling practice remained low despite formal training among many respondents; 76.62 percent identified elimination of cancer cells as the main aim of chemotherapy, 74.10 percent identified intravenous administration as a common route, 35.25 percent always used appropriate personal protective equipment, and 34.89 percent reported that they were well informed about spill cleanup and disposal. Sociodemographic characteristics were not significantly associated with level of knowledge, but several respondent characteristics were significantly associated with safe handling practice. The study recommends mandatory continuing education, current written protocols, reliable personal protective equipment supply, and periodic audit of chemotherapy safety procedures in oncology and related units.