The invention relates to a preparation method of a water-soluble ferrocene supramolecular inclusion compound, belonging to the field of preparation processes of water-soluble metal organic materials in the technical field of supramolecular materials. The preparation method comprises the following steps of: firstly crosslinking beta-cyclodextrin as a raw material with epoxy chloropropane to generate a beta-cyclodextrin polymer under a strong alkaline condition; and then carrying out homogeneous supramolecular reaction on the beta-cyclodextrin polymer as a subject molecule, ferrocene as an object molecule and glycol as a reaction solvent to generate the water-soluble ferrocene supramolecular inclusion compound. The preparation method has the advantages of easy and convenient operation, easycontrol of conditions, little environmental pollution and easy industrialization; the material not only has high water solubility, but also has stable structure without damaging the self structure ofthe ferrocene; and in addition, the water-soluble ferrocene supramolecular inclusion compound can be used in the fields of aqueous-phase electroanalytical chemistry, biological sensors, aqueous-phaseorganic synthesis, biological medicine, and the like because of good water solubility.