Mahkota dewa (Phaleria macrocarpa) has been used as medicinal plant. Garlic, lime, and salt have also been used as flavoring agent for some food products. This research was aimed to utilize mahkota dewa fruit extract combination with garlic, lime, and salt in marinade seasoning as natural preservative agent toward fresh red nila tilapia fish (Oreochormis niloticus). There are 3 extraction methods (maceration and two different types boiling) and 6 different concentrations of extracts (15%, 20%, 25%, 30%, 35%, and 40% of mahkota dewa) with combination of seasoning (10% garlic, 10% lime, and 4% salt) in marinade seasoning were used. The observed parameter included well diffusion method that resulted the best extraction method of mahkota dewa fruit and concentration of extract used in marinade seasoning, where maceration method with 15% and 35% concentration of extract were chosen, which contain alkaloids, tannin, saponin, glycoside, flavonoid, and triterpenoid qualitatively, categorized as low toxic. Moreover, the application to red tilapia fish by microbiology test, descriptive analysis, hedonic test, and challenge test were also conducted. The addition of marinade seasoning with 35% concentration mahkota dewa to fish could preserve the fish until 3 days in refrigerator storage and was accepted in terms of descriptive observation based on SNI, which was proven by challenge test that marinade seasoning have high effectivity toward bacterial growth until 3 days of storage. It was also could improve the taste, aroma, and have potential in longer storage time due to lower moisture content with presence of salt for both fresh and fried fish.