Flying fish roe filament has a high protein content that can be explored as an alternative nutrition food. Unfortunately, those roe filaments still have a fishy odor caused by volatile compounds and fatty acid derivatives. This study aims to eliminate the fishy odor by removing the fat content using the defatting method. The research design used in this study was a simple Randomized Block Design (RDB) consisting of four treatments (control, 1 h, 3 h, and 5 h). The method used in this study for determining amino acids is a descriptive method using HPLC (HighPerformance Liquid Chromatography). The best treatment for extraction time was 5 h with the results of moisture content 10.5 %, ash content 7.5 %, protein content 47.925 %, fat content 8.65 %, total volatile base (TVB) 4.939 %, and yield 86.63 %. TVB value proved that the treatment could reduce the fatty acid content, so the protein concentrate becomes not fishy (from 9.64 to 4.939 %). The best treatment showed the highest content of lysine (149.75 mg 100 g-1) than another essential amino acid.