High-oil maize is those having more than 50% oil content compared with normal maize， and the oil content in embryo accounts for 85% out of the oil in kernel. In order to analyze the development of kernel， embryonic proteome at 15 d （G15）， 25 d （G25） and 35 d （G35） after pollination of high-oil maize inbred line GY220 was isolated by two-dimensional electrophoresis technology， and the isolated proteins were identified by tandem mass spectrometry， followed by analyzing the functions of differentially expressed proteins （DEPs）. As a result， a total of 41 DEPs were identified in three developmental stages. There are 18 up-regulated DEPs in the comparison of G25 with G15， but the down-regulated DEPs was 19. There are 18 up-regulated DEPs in the comparison of G35 with G15， but the down-regulated DEPs was 25. There are 20 up-regulated DEPs in the comparison of G35 with G15， but the down-regulated DEPs was 22. To reveal the functions of the 41 DEPs， GO annotation analysis and KEGG enrichment analysis was performed. The results showed that DEPs were significantly enriched in small molecule metabolism， REDOX and carbohydrate metabolism. Based on the differential protein expression abundance and functional annotation， Zm00001d041962 encoding a glycerol-3-phosphate dehydrogenase and Zm00001d035037 encoding a fructokinase were highly abundant at the stages of 15 d and 25 d， but sharply decreased at the stage of 35 d. Real-time PCR found that the expression of these two genes were accumulated with the embryonic development of GY220， and their orthologs in soybean， peanut and rape shared highly similarity in protein sequences. This study laid a foundation for further improving grain quality and mining functional genes of embryonic development in maize.