Chemotherapy is one of the treatment modalities for cancer. Chemotherapeutic agents had described as carcinogenic, teratogenic, also mutagenic to staff worker so that occupational exposure to chemotherapeutic agents had a highly potential risk. Educational program is a valid tool for providing nurses with updating knowledge and practice about this drugs. Therefore, the aim of this study, examine the effect of implementing educational guidelines about chemotherapeutic drugs on nurse's knowledge and practice. Method and sample, Quasi-experimental research design was conducted in the medical department and outpatient chemotherapy unit at Mansoura Oncology Center of Mansoura University Hospital. The sample of this study composed of 35 nurse who are dealing with this drugs through preparation and administration for cancer patients. Results, indicates significant improvement into nurses knowledge and practice after implementation of educational guidelines. Conclusion, positive effect of educational guidelines about chemotherapeutic drugs on nurse's knowledge and practice.