Docosahexaenoic acid (DHA) is an important ω-3 long chain polyunsaturated fatty acid, which plays important roles in brain development and preventing cardiovascular and cerebrovascular diseases. Phospholipid DHA (PL-DHA) is a form of DHA that has higher bioavailability, which is considered as a potential DHA molecules compared with triglyceride DHA and ethyl ester DHA. This review aims to summarize the recent progress of phospholipid DHA from its effects on human health and preparation methods. Firstly we will introduce the biological activities of DHA throughout human life, after which the absorption process of different structural DHA will be discussed systematically. Then, various preparation methods of PL-DHA, including formulation innovation, extraction, bio-enrichment and enzymatic catalysis, will be summarized in detail. Finally, the future challenges and prospects of PL-DHA will also be discussed.