Euphausia superba has huge biological reserves and much phosphatidylcholine (PC) which is the important phospholipids of growth. However,the research of E.superba is mainly focused on oil and protein,not on PC′s molecules. The study uses Bligh Dyer method assisted with ultrasonic to extract phospholipids from E.superba,optimizing the extraction process by Box-Benhnken design method,and adopted PC molecular shotgun method to identify the PC′s molecules in E.superba by taking choline (m/z 184) as specific pieces. The optimum extraction conditions of phospholipids in E.superba were: ultrasonic time of 21.0 min,ultrasonic temperature of 30 ℃,and solvent dosage of 3.1 mL. Under these conditions,the average actual extraction rate of phospholipids was (7.59±0.03)%. Then,we identified that E.superba contained 36 kinds of phosphatidylcholine molecules by using shotgun method,and the contents of EPA/DHA accounted for 47.75%(269.82 μg·g-1).It is showed that the tissues of E.superba are rich in EPA and DHA,while the omega-3 fatty acid has high utilization efficiency when it exists as phospholipids. The study provides references for the development of nutrition and health care products using E.superba.