This study aimed to investigate the antioxidant potential and bioactive compounds of Pandanus odorifer flower. The phytochemical investigation includes both qualitative and quantitative experiments. In vitro antioxidant studies were conducted using DPPH, ABTS and hydroxyl methods. The bioactive compounds present in the ethanol extract were identified using GC-MS analysis. The MTT assay was carried out on HT-29 colon cancer cells to investigate the cytotoxic ability of the ethanolic extract. The qualitative phytochemical analysis demonstrated the presence of flavonoids, phenolic compounds, tannins, saponins, steroids, carboxylic acids and cardiac glycosides. The determination of flavonoid showed 400 ± 0.110 mg/g and 250±0.090 mg/g of quercetin equivalent in ethanol and water extracts. Similarly, the determination of phenol exhibited 108±0.210 mg/g and 87±0.173 mg/g gallic acid equivalent in ethanol and water extracts. The total antioxidant capacity revealed 1.6 ± 0.093 mg/g and 1.3± 0.056 mg/g equivalent of ascorbic acid in ethanol and water extracts. The DPPH assay showed IC50 values of 35.06 µg/mL in ethanol and 44.82 µg/mL in water extract. Likewise, the ABTS assay revealed an IC50 value of 42.20 µg/mL and 47.45 µg/mL in ethanol and water extract. The hydroxyl radical assay showed IC50 value of 23.95 µg/mL in ethanol and 36.56 µg/mL in water extract. 22 bioactive compounds were identified using GC-MS analysis based on their similarity index. The ethanolic extract showed greater cytotoxic activity with the IC50 value of 28.08 µg/mL on HT-29 colon cancer cells. This study concludes that the Pandanus odorifer flower extracts exhibited significant antioxidant and cytotoxic property.