Introduction: Otitis media with effusion (OME) is a common disease characterized by the retention of fluid and inflammatory by-products in the middle ear without any clinical symptom of acute infection. Objective: To evaluate the level of surfactant in patients with otitis media with effusion. Methods: The surfactant system of the middle ear was examined biochemically in cases of otitis media with effusion and compared to control group. A total of 60 patients with otitis media with effusion compared to 20 normal volunteers. Results: Biochemical analysis of the nasopharyngeal aspirate around the Eustachian tube opening in control group which obtained endoscopically through the nose revealed the presence of phospholipids constituting surfactant with phosphatidylcholine constituting 73.25% of the total phospholipids. Biochemical analysis of the effusion aspirate in cases of otitis media with effusion revealed a highly significant decrease in the total phospholipids compared to normal control group and also a significant change in the phospholipid profile. Conclusion:  significant biochemical changes in the surfactant system of the middle ear are an evident finding in cases of otitis media with effusion. This suggests a possible role for surfactant deficiency in the aetiopathogenesis of cases of otitis media with effusion.