The content of berberine hydrochloride (BH) in compound berberine tablets (CBTs) is subject to strict requirements. Its content is usually measured based on chemical analysis. In this paper, the fluorescence spectral imaging method was used to study the relative content of BH from a physics perspective. By comparing the relative fluorescence intensity of self-made CBTs with different mass percentages of BH, a linear positive relationship was observed between the BH content and the relative fluorescence intensity, and accordingly the quality of CBTs of different brands was evaluated. The results indicate that the fluorescence spectral imaging method can be a simple, fast and nondestructive semi-quantitative analysis method to determine the content of BH in CBTs, and this method has great potential in the quality control of CBTs.