Meibomian gland dysfunction (MGD) is a common ocular surface disease, characterized by terminal duct obstruction and (or) qualitative and quantitative changes in the glandular secretion. Meibomian lipids are a mixture consisting of various lipids, and they form the lipid layer of tear film and play important roles in preventing the evaporation and maintaining the stability of tear film. Patients with MGD may have different compositions of meibomian lipids, which could be an important indicator for diagnosis of MGD. The methods to analyze the composition of meibomian lipids mainly include chromatography and spectrum analysis. Because the pathogenic factors of MGD are still unclear, the main treatment is to relieve the symptoms. It can contribute to diagnose and treat MGD if we can find some convenient and effective methods to analyze the quality and quantity of meibomian lipids.