The invention discloses a preparation method of an ASP flooding crude oil demulsifier, which comprises the following steps: (1) mixing tert-butyl phenol and triethylene tetramine, heating until complete dissolution, and dropwisely adding a formaldehyde solution to generate an initiator A; (2) putting the initiator A and a catalyst into a reaction kettle, sealing, vacuumizing, introducing epoxypropane, heating, and controlling the pressure to obtain an intermediate product B; (3) putting the intermediate product B and a catalyst into a reaction kettle, sealing, vacuumizing, introducing epoxypropane, heating, and controlling the pressure to obtain polyether C; and (4) adding polyether C into the reaction container, dissolving polyether C, adding choline chloride, stirring, heating to 50-75 DEG C, completely dissolving, dropwise adding TDI, and finishing dropwise adding when the wire drawing length is 3-4cm, thereby obtaining the ASP flooding crude oil demulsifier. The demulsifier obtained by the process has good lipophilicity, can significantly improve the use efficiency of a chemical agent, and has high surface activity, high demulsification speed and good demulsification effect.