Abstract: Objective:To identify and activity screen of peptides in Apostichopus japonicus oligopeptides. Methods:The peptides in Apostichopus japonicus oligopeptides and their source proteins were analyzed and identified by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). The antihypertensive peptides were screened by online database and structure-activity relationship. Results:88 small peptides in Apostichopus japonicus oligopeptides were identified,containing a large number of unpublished small bioactive peptides. The identified peptides were derived from 19 kinds of Apostichopus japonicus proteins,mainly containing collagen,putative collagen alpha-1(I)chain,2 alpha fibrillar collagen,putative glycine-rich protein and extracellular matrix protein 3,etc. 9 antihypertensive peptides were obtained by online database activity screening. Further combined structure-activity relationship analysis,the core peptides KFPPPM,WEPPTFDGGRP,NPSPPF,FDGPEGPR and RPQYPQYPS had potential antihypertensive activities and were expected to be developed as antihypertensive peptide medicine. Conclusion:This method is simple,rapid,high throughput and high efficiency for analysis and identification of multiple peptides in Apostichopus japonicus oligopeptides. It realizes the rapid identification of multiple peptides in complex matrix samples and high-efficiency activity screening. It has potential application value.