The invention discloses a membrane-free direct alcohol fuel cell in an acid solution and a preparation method of the membrane-free direct alcohol fuel cell. The membrane-free direct alcohol fuel cell comprises a gas diffusion electrode, an anode strip and electrolyte, wherein the gas diffusion electrode is formed by hot pressing a catalyst layer, a stainless steel wire and a water resistible and breathable layer which are sequentially stacked, the water resistible and breathable layer is directly contacted with air, a plastic gridded clapboard is arranged between the gas diffusion electrode and the anode strip, rubber gaskets are arranged between the plastic gridded clapboard and the anode strip as well as between the plastic gridded clapboard and the catalyst layer, the catalyst layer is manufactured by a Pd-Co-Cu nanocomposite supported by a multiwalled carbon nanotube, the anode strip is formed by taking Nafion solution as a binder, and coating the stainless steel wire by Pt/C partials, and the electrolyte is a sulfuric acid solution containing alcohol. The membrane-free direct alcohol fuel cell can stably discharge in the acid solution, an ion exchange membrane is not used, the membrane-free direct alcohol fuel cell is simple in structure, and the cost of the membrane-free direct alcohol fuel cell is greatly reduced.