Docosahexanoic acid (DHA) is one of the essential omega-3 fatty acids for humans for the development of brain tissue and the nervous system. Tiger prawn have a larger body shape with a relatively much more expensive price than vaname shrimp. All physical criteria and market prices are not necessarily a parameter that DHA levels in tiger prawn are higher than vaname shrimp. This study aims to determine the levels of DHA contained in vaname shrimp and tiger prawn. The gas chromatography conditions used were programmed temperature with an initial column temperature of 130°C, a temperature increase of 2°C/minute to 230°C (held for 20 minutes), using hydrogen as a carrier gas with a flow rate of 10 mL/minute. The method was linear with a correlation coefficient of 0.9993, within a concentration range of 312.5 - 3125 ppm. The limit of detection (LoD) and limit of quantitation (LoQ) of DHA were 97.35 ppm and 324.5 ppm, respectively. DHA retention time was at 54.058 minutes. The results showed that the fat content in vaname shrimp was 28.7 ± 1.07 mg/g and tiger prawn was 29 ± 1.64 mg/g. DHA levels in vaname shrimp amounted to 0.62 ± 0.005 mg/g and tiger prawn amounted to 0.275 ± 0.01 mg/g.