The objective of the present work was to develop a solid-phase extraction-gas chromatography (SPE-GC) method to isolate and measure free fatty acids (FFA) in lipid-rich seeds for immediate use in our investigations on the non-orthodox storage behaviour of numerous tropical seeds, such as Citrus and coffee seeds, which generally contain large amounts of lipids. In order to develop a sensitive and reliable method, two SPE reference procedures (1,3) employed in food chemistry were compared using a 100/1 mixture of triolein/heptadecanoic acid. The SPE method providing the best results, together with the fatty acids (FA) methylation procedure were then further refined for decreasing triacylglycerols (TAG) contamination in the FFA fraction to achieve accurate measurement of FFA in lipid-rich seeds. The method was finally compared to the conventional thin layer chromatography (TLC) purification procedure used classically in plant physiology studies and validated in ageing coffee seeds.