The inclusion phenomena of β-Cyclodextrin and Alizarin complexone was investigated by 1H NMR, 2D ROESY, UV-Visible Spectroscopy, Fluorescence Spectroscopy, HRMS and SEM study. The change in chemical shift values in 1H NMR study established the possibility of interactions between the corresponding protons of β-Cyclodextrin and AC molecule, which was further confirmed by the appearance of cross peaks in 2D ROESY spectrum. The Job plot method gave the host-guest stoitiometric ratio to be 1 : 1. The association constant and thermodynamic parameters like ΔH 0 , ΔS 0 and ΔG 0 were calculated from UV-Visible spectroscopy. The experimental results showed, the fluorescence intensity of AC enhanced with increasing concentration of β-Cyclodextrin. The plot of 1/(F-F 0 ) against 1/[β-Cyclodextrin] gave a straight line indicating 1 : 1 stoichiometric ratio of the inclusion complex formed. In this inclusion process, the ΔG < 0 value describes the process to be a spontaneous one.