In this study, lipid classes, fatty acid composition, and molecular species of betaine lipid of the green algae Halimeda incrassata Lamx., collected from Truong Sa archipelago, have been identified. Five lipid classes including Pol, ST, FFA, TG, HW have detected on TLC chromatography. In the total lipid, 21 fatty acids have been indicated, in which, 4 fatty acids belonging to the PUFAs group with the valuable bioactivities and the high content are ALA, GLA, AA, EPA. From Halimeda incrassata, this is the first time 17 betaine lipid molecular species have been identified by HPLC‐HRMS technique. Interestingly, all of these molecular species belong to the diacylglyceryl‐N,N,N‐trimethylhomoserine group, which is the characteristic composition of the non‐flower plant.