An ischemic stroke occurs when the blood supply to part of the brain is interrupted or reduced, preventing brain tissue from getting oxygen and nutrients. Brain cells begin to die in minutes. Ischemic stroke can be treated by using Antiplatelet drug such as aspirin as an immediate treatment which may prevent blood clots and improve recovery after stroke. Glycerol is also used as it decreases the edema in ischemic stroke. The aim of the present investigation is to develop a novel formulation of oil in Glycerol emulsion comprising aspirin in the treatment of ischemic stroke and in vitro evaluation. The oil in glycerol emulsion was prepared by high shear homogenization and prepared emulsions were evaluated for particle size, drug content, in vitro drug release and stability. Based on log P values of aspirin, olive oil is selected as oil phase and tween 80 was used as emulsifying agent. Total 10 formulations F1to F10 were prepared and among these, in F1-F6 phase separation was observed within 24 hrs whereas F8 to F10 were stable for one week and later creaming was observed and it is re-dispersible. The particle size was determined by optical microscopy and the average particle size for F8, F9 and F10 were 6.04, 3.38 and 7.8 µm respectively. Drug content was determined as 68±3.95%,108±5.816%, 103±4.61% respectively in compositions F8, F9 and F10. Rate of drug release followed first order kinetics and an immediate burst release effect was observed within 15 minutes. It is concluded that olive oil and glycerol emulsions containing aspirin is prepared successfully and evaluated. It is a promising formulation approach for co administration of aspirin and glycerol in the treatment of ischemic stroke. The formulations were physically and chemically stable. In vivo application of this formulation will be done in suitable animal models in future.