This paper reports a discrimination study based on the physico-chemical characteristics, fatty acids and profile of volatile compounds of the seeds from seven date palm varieties (Phoenix dactylifera L.) grown in Tunisia. Date seeds contained 10.49–14.76% moisture; 6.28–11.2% fat (on a dry weight basis); 2.67–12.85% protein; 0.91–6.06% reducing sugar; 0.61–2.98% sucrose and 0.97–1.17% ash. Gas liquid chromatography revealed that the oil fraction of the date palm seeds contained eighteen fatty acids, with oleic acid (30.77–42.50%) and lauric acid (18.51–27.48%) as the main unsaturated and saturated ones. Volatile profile showed differences among varieties. In total, forty-five compounds were identified, mainly alcohols, aldehydes and unsaturated hydrocarbons ones. This study provides evidence that the seeds of date may be a potential source of valuable nutrients with interesting functionality.