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The present study was conducted to examine various biological activities of a methanol extract of <em>Agave attenuata</em> leaves. GC-MS analysis of the <em>n</em>-hexane fraction from the extract revealed the presence of 31 compounds, with mono-2-ethylhexyl phthalate (11.37%), 1,2-benzenedicarboxylic acid (6.33%), <em>n</em>-docosane (6.30%) and eicosane (6.02%) as the major components. The leaves contained appreciable levels of total phenolic contents (10.541–39.35 GAE, mg/100 g) and total flavonoid contents (43.35–304.8 CE, mg/100 g). The extract and some of its fractions showed moderate antimicrobial effects. Leaves extract and fractions also exhibited a good antioxidant potential when measured by DPPH radical scavenging activity and inhibition of lipid peroxidation assays. The hemolytic effect of the plant was found to be in a range of 1.01%–2.64%.<strong> </strong>From the present study it is concluded that this plant could be used as a source of natural antioxidants and functional food nutraceutical applications.