Brown macroalgae represent a sustainable and abundant source of lipids with acknowledged functional and health benefits. Nonetheless, macroalgae lipidome has been poorly unraveled due to lipids complex structural and chemical diversity. In this study, a comprehensive lipidomic analysis was performed in four macroalgae: Saccharina latissima, Fucus vesiculosus, Fucus serratus and the invasive Sargassum muticum, using HILIC-C30RP-HRMS. Neutral lipids (tri-, di-glycerides) comprised 72–82% of total lipids (TL) with a highly unsaturation profile (27–49% depending on species). The polar lipidome comprised glycolipids, phospholipids, betaine lipids and sphingolipids with varied content among macroalgae. S. latissima displayed the greatest level of glycolipids (23% of TL), by contrast with the dominance of long-chain polyunsaturated betaine lipids (10–18% of TL) in the other species, particularly in S. muticum. Phospholipids and sphingolipids were detected in low abundance (<1.7% of TL). This study elevated the potential of brown macroalgae as an emerging reservoir of bioactive lipids with nutritional relevance.