Despite the discovery of many plant-based drugs, research is still ingoing, in addition to using safer and more effective methods in the extraction process. Presently, Super Fluid Extraction (SFE) was applied to extract the mango seed kernels (MSK) at two operating temperatures 30 and 50 °C with studying their biological activities. MSK extract via SFE at operating temperatures 30 °C showed high yield 0.268 g compared to that at 50 °C. High-performance liquid chromatography (HPLC) analysis demonstrated high content of compounds such as gallic acid (68503.01 µg/mL), chlorogenic acid (6541.68 µg/mL), ferulic acid (2555.12 µg/mL) and ellagic acid (2479.42 µg/mL) MSK while at 50 °C the concentrations were low. Helicobacter pylori was highly inhibited by MSK extract via SFE at operating temperature 30 °C with inhibition zone 29.17 ± 0.29 mm, MIC 15.62 µg/mL and MBC 31.25 µg/mL, while at operating temperature 50 °C the inhibition zone was 24.67 ± 0.58 mm, MIC was 62.5 µg/mL and MBC was 125 µg/mL. Moreover, low hemolysis was recorded by H. pylori treated by MSK extract via SFE at operating temperature 30 °C compared to that at 50 °C. Excellent antioxidant with IC