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Abstract Phoenix dactylifera L. (date palm) is one of the most valued crops worldwide for its several industrial and nutraceutical applications. The date fruit (pericarp) has served as an essential food component in the hot arid regions owing to its nutritional value. Currently date pit, considered as a waste product, is employed as an animal feed, and more recently as coffee substitute post roasting. In contrast, pollen is another valuable date by-product used as dietary supplement for enhancing male and female fertility. In this study, a large-scale comparative metabolomics approach was performed first time, for the characterization and standardization of date palm (variety Majdool) by-products via a multiplex approach combining SPME, GCMS, UPLC-MS and NMR spectrometry. Moreover, roasting impact on date pit metabolite composition was also assessed using same analytical platforms. SPME headspace analysis revealed for the abundance of furans, pyrans, terpenes and sulfur compounds in roasted date pit, whereas pyrroles and caffeine reported in coffee were absent. GCMS revealed for fatty acids enrichment in roasted pit versus sugars abundance in roasted coffee. The results justified for the use of roasted pit as a potent antioxidant and caffeine-free coffee substitute. LCMS and NMR analyses identified many chemically diverse primary and secondary metabolites in date pit and pollen viz., phenolic acids, flavonol conjugates, fatty acids, sphingolipids, steroids and saponins. The enrichment of steroids in date pollen could be responsible for its fertility effects |