Aims: to assess patient caregiver's knowledge and practice toward safe handling chemotherapy, development of suggesting guidelines for patient caregivers toward safe handling chemotherapy. Methods: a study was conducted in oncology unit at Assiut University hospitals. To accomplish the purpose of this study data were collected from all patient caregiver in oncology a convenience sample of all available patient caregivers about (60) male and female working in oncology department who were participated in this study, who are currently assigned and provide care for adult patient with chemotherapy, in period from preparation until 72 hour post administration, it consist of (20) nurses, (20) worker and (20) patient family member. Tool utilize for data collection were an Arabic structured interview questionnaire sheet for patient caregivers and observation checklist for patient caregivers. Results: the result implies that there was lack of nurse's knowledge about chemotherapy with percentage (95%).In addition workers had low knowledge about chemotherapy with percentage (50%, 85%, 65%and 45% respectively).Family member had low knowledge regarding knowledge about cancer and chemotherapy at high percentage of score with percentage (85%, 100%and 80 respectively). And there is lack of practice related to there are some equipment are not applicable (use ducted bio-safety cabinet, a long sleeved gown, safety spectacles or goggles, spill kit).Conclusions: recommendation, further research studies are needed to focus on more training and education for patient caregivers toward safe handling chemotherapy and health monitoring for health care worker to prevent or reduce risk of exposure.