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This study assessed the metabolite content and bioactivities of Kakadu plum (KP) from Western Australia (WA). LC-MS/MS and UHPLC-PDA analyzed sugar, vitamin C, and ellagic acid (EA). Functional properties were evaluated by spectroscopic technique, agar well diffusion, and microplate dilution methods. WA KP exhibited higher total sugar (16.3 ± 1.0 g/100 g DW) and free ellagic acid (EA) (23.2 ± 1.7 mg/g DW), along with abundant vitamin C (25.20 ± 0.16 to 131.50 ± 0.20 mg/g DW) compared to Northern Territory KP fruits. The fruit showed strong antioxidant activities, α-glucosidase inhibition, and effectiveness against bacteria, with positive correlations to total phenolic content (TPC), vitamin C, and EA. These findings highlight WA KP’s potential for functional foods and pharmaceuticals, emphasizing the importance of TPC, vitamin C, and EA in selecting high-quality fruit.