Marine macroalgae are emerging as sustainable and healthy food, and a source of lipids with nutritional and bioactive properties. Nonetheless, the lipidome of macroalgae is little explored compared to other photosynthetic organisms. Most of the literature is devoted to the analysis of fatty acid (FA) from macroalgae, because of they are a source of long-chain omega-3 and omega-6 polyunsaturated FA, which can be used as an alternative to fish oil. However, FA occurs mostly esterified in other lipids, such as polar lipids, that still overlooked. The data published on the study of polar lipids (phospholipids, glycolipids, betaine lipids) pinpoint their remarkable characteristics and bioactive properties which mainly remain unexplored. The lipidome of macroalgae seems to be species-specific, while some lipid classes appear to be characteristic of phyla. Lipidome plasticity of macroalgae is verified to change under abiotic and biotic factors which may modulate the functionality and add-value of the biomass. This biochemical plasticity represents an advantage to obtain ingredients for target applications and niche of markets. This review provides an overview on macroalgae lipidome characterization, addressing analysis workflow based on liquid chromatography coupled to mass spectrometry, and discussing the bioactive properties and biotechnological applications of macroalga lipids. Practical Applications: The current state of the art on marine macroalgae lipidomics is reviewed, including the lipid characterization using modern liquid chromatography-mass spectrometry, the distribution of polar lipids by macroalgae species and phylum, their bioactive properties, and potential biotechnological applications. In a time of increasing awareness of sustainably produced marine resources, the lipids from macroalgae are still underexplored. This review provides information for smart valorization of macroalgae as source of add-value lipids contributing to the development of blue bioeconomy.