The research related by evaluate the fatty acid composition of durian seed oil by Gas Chromatography combined with Mass Spectrometry (GCMS) and analysis stereospecific numbering of Fatty Acid Triglycerides (FAG). The extraction was performed by maceration process using nhexane solvent. Faster Adoption and Manufacturing (FAME) from durian seed was synthesized using concentrated sulfuric acid as acid catalyst and analyzed using GC-MS and Fourier Transform Infra-Red (FTIR) spectroscopy. The results obtained of methyl ester synthesis were 10- octadecadienoic acid methyl ester, octadecanoic acid methyl ester, arachidonic acid methyl ester. Stereo specific numbering of FAG was analyzed by liquid gas chromatography Flame Ionization Detector (FID). The results showed that the major component of fatty acid triglyceride was C50-POP 17.5878%; C54-SLS 14,5857% and C50-PSP 12,7259%. The position of triglyceride seed oil of saturated fatty acid durians is generally on sn-1 and sn-3 whereas unsaturated fatty acids in sn-2.