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Background: Propolis is a bee (Apis mellifera) product of plant origin with varied chemical composition depending on the ecology of the botanical origin. It has been reported in literature to possess various therapeutic effects both traditionally, clinical trial, and animal study., Objectives: In the present study bioactive principle in methanol extract of Nigerian bee (A. mellifera) propolis was determined by gas chromatography-mass spectrometry (GC/MS) study., Materials and Methods: The methanol extract of Nigerian bee (A. mellifera) propolis was characterized for its chemical composition by preliminary phytochemicals screening and GC/MS analysis using standard procedures and methods., Results: Phytochemical screening revealed the presence of flavonoids, saponins, alkaloids, tannins, cardiac glycosides, anthraquinones phlobatannins, and steroids while GC/MS chromatogram revealed nineteen peaks representing 60 different chemical compounds. The first compounds identified with less retention time (RT) (13.33s) were methyl tetradecanoate, tridecanoic acid, methyl ester, decanoic acid, methyl ester while squalene, all-trans-squalene, 2,6,10-dodecatrien-1-ol, 3,7,11-trimethyl-, (E,E)- and farnesol isomer a took longest RT (23.647s) to identify. Methyl 14-methylpentadecanoate, hexadecanoic acid methyl ester, methyl isoheptadecanoate, and methyl tridecanoate were the most concentrated constituent as revealed by there peak height (26.01%) while eicosanoic acid was the least concentrated (peak height 0.81%) constituent of Nigerian bee propolis., Conclusion: The presence of these chemical principles is an indication that methanol extract of Nigeria bee propolis, if properly screened could yield a drug of pharmaceutical importance.