ABSTRACT: This study was aimed at characterizing four Tunisian pistachio cultivations (Pistacia vera) from the Mateur (North), Nabeul (North-East), Kairouan (Middle) and Sfax (Middle-East) regions and two varieties, Mateur and Ohadi, considering fatty acid composition and main lipid class contents (polar lipids, diacylglycerols, triacylglycerols and free fatty acids) in ripe seeds. Lipid classes were separated using thin layer chromatography, and fatty acid methyl esters (FAMEs) were analyzed by gas chromatography.