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Phytochemicals (PCs) are gaining popularity due to their antioxidant effects and potential protection against infection, cardiovascular disease, and cellular metabolic activity. These PCs must be retained as much as possible during extraction. This research focused on the extraction of PC from Psidium guajava Linn. leaves due to higher antioxidant potential. Solvent extraction (SE), microwave-assisted extraction (MAE), and ultrasound-assisted extraction (UAE) using distilled water (DW) or 60% (v/v) ethanol/water (ET) were used for the extraction of PC. ET shows higher total phenolic (TPC) and total flavonoid content (TFC) as well as higher antioxidant activity than DW. Phytochemical screening demonstrated that all of the screening showed positive results in all extraction methods, except glycoside. There were no significant differences (p > 0.05) in TPC and TFC during MAE/ET, SE/ET, and UAE/ET. Antioxidant analysis shows that MAE and SE resulted in high (p < 0.05) DPPH and FRAP values for ET and DW, respectively. MAE/ET showed the highest inhibitory activity (IC₅₀ = 16.67 µg/mL). HPLC and TLC analysis reveal the fingerprint of morin, which might function as an anticancer agent with other bioactives. Increasing the extract content increased the inhibitory activity of SW480 cells via MTT assay. In conclusion, MAE/ET is the most efficient among the extraction techniques in terms of anti-cytotoxicity effects.