The invention provides a weeding composition containing glufosinate-ammonium and fluoroglycofen-ethyl, and a preparation method of the composition, and belongs to the technical field of pesticides. The dosage form of the composition is a micro-emulsion, the composition comprises glufosinate-ammonium, fluoroglycofen-ethyl, a cosolvent, a first surfactant and a second surfactant, the first surfactant is alkyl glycoside, and the second surfactant is polyoxyethylene phosphonate or polyoxyethylene phosphonate salt. The weeding composition prepared in the invention has the characteristics of fast weeding speed, good weeding effect, low production cost and environmental protection, and has an obvious better weeding effect than single doses of glufosinate-ammonium and fluoroglycofen-ethyl. The preparation method has the advantages of simplicity, convenience in operation, and high feasibility; and the above product has stable quality, still has good quality stability after being stored for a long term, has obvious market values, and is suitable for large-scale promotion application.