Breastfeeding support from healthcare professionals influences a mother’s decision to initiate and maintain exclusive breastfeeding. Objectives: To assess the level of knowledge on exclusive breastfeeding (EBF) among healthcare professionals in a ljalaa maternity hospital in Tripoli –Libya. Materials and methods: Across-sectional study was performed between March to April 2021. Study subjects included health professionals working in the Aljalaa maternity hospital. A questionnaire was used to collect data. SPSS 22 package program was used for statistical analysis. Results: A total of 118 health care professionals participated in this study. The majority of the participants52.5% was nurses and midwives, 86.4% were females, with mean age 37.43±9.37 and 82.2% were resident with more than ten years of experience. Health care professionals had adequate knowledge about BF initiation, duration and complementary feed. However, 78.8%% of the providers didn’t know about the frequency of breastfeeding (BF). The majority of the participants knew about breast milk sustainabilitybut,81.4% didn’t know about cracked nipple management. Health care professionals have good information about BF contraindications, the majority75.4% of the providers answered diarrhea is an indication to stop breastfeeding, and 65.3% answered small breasts affect breastfeeding. The mean score of the professional’s healthcare knowledge was 54.36 ±13. Conclusion: The level of breastfeeding knowledge of health care professionals was fair with deficits in the important key areas. Immediate training and continuing medical education for health workers will help improve mother and newborn health.