The invention discloses a preparation method of a duel-alkylphenol polyamine emulsifier, prepared emulsified asphalt and application thereof. The duel-alkylphenol polyamine emulsifier preparation method comprises the following steps of: (1) adding alkylphenol, polyethylene polyamine, a solvent and a catalyst into a reaction container to form a reaction system, heating, and dripping an aldehyde to react so as to obtain a midbody; (2) cooling the midbody to 0-25 DEG C, adding the alkylphenol and the polyethylene polyamine, heating again, dripping the aldehyde, after the reaction is completed, and distilling out the solvent, thereby obtaining a target product. As biological enzyme is adopted as the catalyst for synthesizing the emulsifier, the preparation method disclosed by the invention is high in reaction activity and environmentally friendly, and meawhile the prepared duel-alkylphenol polyamine emulsifier is good in adaptability with aggregates, and the water damage resistance of the aggregates can be improved. The raw materials used in the preparation process of the preparation method for the duel-alkylphenol polyamine emulsifier disclosed by the invention are all easy to obtain, no rigorous preparation conditions are needed in the process of synthesizing the emulsifier, the heating temperature is not very high, the yield is relatively high, and the synthesis process is relatively simple to operate.