In this study, the crude rapeseed peptide was used as the research object. After treatment with powdered activated carbon, peptide solution was adsorbed onto DA201-C macroporous adsorption resin and then desorbed by ethanol to obtain rapeseed refined peptides (RP75). The basic composition, antithrombotic activity, anti-inflammatory activity and amino acid composition of rapeseed refined peptides (RP75) were also analyzed. The results showed that the optimum process for the refining of the DA201-C macroporous adsorption resin were peptide concentration 30 mg/mL, flow rate 3 BV/h, pH 4.5 and desorption concentration of the ethanol 75%. Peptide content was 74.94%, no glucosinolate or phytic acid was detected in RP75. On the other hand, RP75 contained rich amino acids and exhibited excellent bioactivities including anti-thrombotic and anti-inflammatory activities with IC₅₀ values of 8.82 mg/mL and 0.80 mg/mL. These results indicated that RP75 could be used as a potential health promoting ingredient with functional activity.