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The reaction of allyl isothiocyanate (AITC) with hydroxyl/water and β-cyclodextrin (β-CD) in different acidic-alkaline media has been investigated by ultraviolet spectrometry. The kinetic parameters of the reaction were measured. It was found that after AITC translating into thiourea, the absorption peak shifted from 240 to 226 nm and the molar absorptivity increased about 16 times. The reaction can be seen as a pseudo first order reaction because the concentration of hydroxyl was constant. β-CD can inhibit the reaction of AITC with hydroxyl/water, i.e. the hydrolysis of AITC. The formation constant (Ka) and thermodynamic parameters of the complex reaction were calculated. Ka decreased with the increase of temperature. The experimental results indicated that the inclusive process was an exothermic and enthalpy-driven process accompanied with a negative entropic contribution.