The invention discloses a multi-sulfo surfactant as well as a preparation method and application thereof. The method comprises the following steps: 1) dissolving dipentaerythritol in a first solvent,adding triethylamine, stirring and heating to 50-60 DEG C, dropwise adding perfluorovaleryl fluoride while stirring, continuously heating to 75-85 DEG C, stirring and reacting for 4-6 hours, washing the product, and performing extraction to obtain a faint yellow solid; 2) dissolving the faint yellow solid in a second solvent, adding sodium hydroxide after the faint yellow solid is completely dissolved, stirring and reacting at 25-35 DEG C for 0.5-1 hour, adding a sulfonate solution, heating to 75-85 DEG C, carrying out heat preservation for a period of time, and carrying out suction filtrationon a reaction product to obtain a solid, namely a product; and 3) adding a catalyst and a third solvent into the obtained solid product, dropwise adding diisocyanate, reacting for 3-5 hours at the reaction temperature of 40-60 DEG C, and drying in vacuum to obtain the multi-sulfo surfactant. The surfactant solves the problem that the surface activity is not high enough due to short fluorocarbon fragments, and the surfactant is used as a chemical oil displacement agent to improve the recovery ratio of crude oil in tertiary oil recovery.