Objective In Tanzania, seminars and training on the preparation and administration of injectable artesunate were given to health care providers (HCP) during its introduction in 2013. Published evidence on knowledge of its preparation among HCP in public health facilities is scarce. The study determined level of knowledge of health care providers (HCP) in public health facilities on the preparation of injectable artesunate for severe malaria. Results Most (82.5%) of HCP had low knowledge on preparation of injectable artesunate; 78.8% of HCP did not know what to do if the mixture do not mix-up correctly, 73.7% did not know when to discard the preparation if not used and half (50.1%) of them knew how to correctly calculate the dose based on body weight.