Epilobium angustifolium is widely used in medicine for disease treatments, as well as in the cosmetic and food industries. The aim of this research was to investigate the antioxidant, and anticholinergic properties, phenolics profile and antibacterial activities of the E. angustifolium ethanol extract. The analysis of phenolic compounds was performed with LC-MS/MS. The antioxidant capacity (radical scavenging, metal-reducing power and total antioxidant activity) was assessed by DPPH, ABTS, Cu2+-Cu+ reducing (CUPRAC), Fe3+-Fe2+ reducing and ferric thiocyanate methods. The antibacterial activity was determined by disc diffusion and MIC (Minimum inhibitory concentration) methods and the anticholinergic property was predicted by inhibition of acetylcholinesterase (AChE). The major phenolic compounds, founding in the plant extract were luteolin, fumaric acid, vanillic acid, and caffeic acid. The ethanol extract of the plant showed DPPH free radical scavenging value of 11.3%, while the ABTS radical scavenging activity was 19.4% and showed moderately metal-reducing power. Also, the extract had 39.3% inhibition on lipid peroxidation of linoleic acid emulsion and showed an inhibition effect on the AChE with IC50 values (0.14 mg mL(-1)). The ethanol extract of the plant showed antibacterial effect on Staphylococcus aureus, Escherichia coli, and Salmonella Typhimurium at different levels. These results suggested that E. angustifolium extract might be a suitable natural antioxidant in the preservation of foods by preventing the oxidation of polyunsaturated fatty acids, and might play a role in the treatment of some diseases with its antioxidant, anticholinergic, and antibacterial activity.