Abstract: In this paper,corn gluten meal was used as experimental material. High Fischer ratio peptide (HFRP) derived from corn protein was prepared by complex enzymatic hydrolysis (alkaline protease enzyme,flavourzyme). Further purification and structural identification were carried out by different molecular,charge and hydrophobicity. And intestinal absorption and resistance to gastrointestinal digestion were explored. Results showed that HFRP with the F value of 26.29 was obtained by complex enzymatic hydrolysis using corn gluten. Twelve components were separated and collected by different molecular,charge and hydrophobicity. By building the Caco-2 cells absorption model to study the absorption status of different characteristics components,four short peptides of high absorption with mass/charge ratio of 203.1409,229.2980,226.9517,284.2966 which were LA,IP,PL and HQ had identified by mass spectrometry. By studying the gastrointestinal digestion of different characteristics components,two short peptides of resistance to digestion with mass/charge error of 226.9517,284.2966 which were PL and HQ were identified by mass spectrometry. PL and HQ were small molecular weight,high degree of hydrophobicity,positive charge,preferential absorption and resistant digestion. This study could provide an experimental basis for the development and utilization of HFRP derived from corn protein.