With the development of the times, there are more and more researches on 4-Methylumbelliferone (4-MU). Now the problem of 4-MU research is that the water solubility of 4-MU is poor, leading to its low bioavailability. It is necessary to improve the water solubility of 4-MU by β-Cyclodextrin (β-CD) inclusion complex prepared by computer hydrogen spectroscopy technology. In this paper, the optimum preparation process was selected by an orthogonal test. 4-Methylumbelliferone-β-Cyclodextrin inclusion complex (MCIC) was characterized by computer hydrogen spectroscopy using ultraviolet spectrum, infrared spectrum, X-ray diffraction and nuclear magnetic resonance hydrogen spectrum. And its solubility was determined. The results showed that MCIC was confirmed by ultraviolet spectrum, infrared spectrum, X-ray diffraction and hydrogen magnetic resonance spectroscopy. The solubility test showed that the water solubility of 4-MU increased by nearly 20 times after the inclusion complex was formed. Therefore, it can be concluded that the preparation process of MCIC based on computer hydrogen spectrum technology is simple, cheap and stable. The solubility and bioavailability of 4-MU were significantly improved by the preparation of the inclusion complex.