Spectrophotometric method is widely used in the structure determination of biologic macromolecules and non-covalent interactions study for its convenience and speed.In the present paper,spectroscopy methodologies in the study of non-covalent interactions between small-molecule and biomacromolecule is comprehensively reviewed with 25 references．This review article focuses on the applications and development of common spectroscopy methodologies in the study of non-covalent interactions between small molecule and biomacromolecule,including the UV,fluorescence,CD,IR,Raman,resonance light scattering technique and SPR.The advantages and disadvantages of spectroscopy methodologies are also described.UV-Vis absorption spectrum (UV) method is widely used in the study of non-covalent interactions for its convenience and speed.The binding site number,the apparent binding constant and the interaction mode of non-covalent interactions can be obtained by fluorescence spectrum method.Circular dichroism (CD) method is effective way in the study of non-covalent interactions measure.Spectroscopy information about protein secondary structure and conformation can be acquired by infrared spectrometry (IR) method.Raman spectroscopy method is a better way to investigate the conformation change in macromolecules in solution.Non-covalent interactions can be measured by surface plasma resonance (SPR) method under the natural active condition.X-ray diffraction analysis method is better for non-covalent interactions research,but it is difficult to cultivate crystalline complex.