Objective To establish an experimental method for determining the glycemic index (GI) in honey-containing herbal preparations in mice. Methods With reference to the WS / T 652-2019 standard, the experimental method for mouse testing was optimised. Samples comprising fructose / glucose syrups at varying ratios were employed to validate the feasibility of the preliminary conditions. The optimised method was applied to the determination of the GI of honey and the traditional Chinese medicinal preparation containing honey-Guhan Nutritive Essence. Results Under the optimized conditions, the GI values of fructose / glucose syrup were closest to the theoretical values, and the correlation with the fructose ratio was highly linear (R²= 0. 9959). The GI values of the four honeys with different fructose / glucose ratios were 60. 3, 54. 3, 56. 5, and 59. 3, respectively, which were lower than those of fructose / glucose syrups with the same ratios. The GI values of the honey-containing herbal preparation Guhan Nutritive Sample ranged from 45. 2 to 59. 6, and decreased with decreasing honey GI. Conclusions This optimized mouse assay is expected to provide an effective tool for determining the GI of honey-containing herbal preparations and for screening honey raw materials.