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Barleria dinteri is a medicinal plant with distribution in the Limpopo, Gauteng and Mpumalanga provinces of South Africa as well as in Botswana, Swaziland and Namibia with exclusive specific habitation on limestone-rich soil. The plant species is used by traditional healers for wound healing and treatment of some intestinal tumours, as well as to relieve joint pains and toothache. The present study was aimed at the metabolite profiling of the different tissues (branches, flowers, leaves, roots) of Barleria dinteri using GC-MS analysis. Different extracts of the plant parts samples were subjected to GC-MS analysis and detected compounds were compared for presence amongst the different tissues. The results of the study revealed that all different parts (branches, flowers, leaves and roots) of B. dinteri, possess compounds that are detectable through GC-MS analysis with most compounds detected in the aerial parts, particularly the flowers. The results of the current study could serve as a basis for the possible plant parts substitution of the roots of B. dinteri with the aerial parts and the exploration of the pharmacological properties of the flowers for sustainable uses of the plant species for medicinal purposes.