The invention discloses an omega-vinylsulphonyl disulfide compound as well as a preparation method and an application thereof. The compound has a structure with a structural formula I described in the specification, wherein n is an integer of 4-18. The compound can form a highly-ordered self-assembled film on the surfaces of noble metals such as gold and silver, wherein a vinylsulphonyl group on the surface of the self-assembled film can respectively react with biological/chemical molecules that contain sulfydryl, amino and hydroxyl under a certain condition for molecular coupling, biological active material preparation, biochemical sensor chip modification, and the like. The self-assembled film prepared by the compound has the advantages of hydrolysis resistance, alkali resistance, and response spectrum resistance, and the like.