The invention belongs to the technical field of clinical in vitro diagnostic reagents and relates to a dry chemical reagent tablet for quantitatively measuring the concentration of total cholesterol and a preparation method of the dry chemical reagent tablet. The dry chemical reagent tablet comprises four laminations which are respectively a support layer, a color developing layer, a reagent layerand a diffusion layer distributed sequentially; and reagents in the reagent layer include a buffer system, a hydrophilic colloid, peroxidase, an activator, a surfactant and a stabilizer; and reagentsin the color developing layer include a chromogen and a hydrophilic colloid. Compared with the prior art, the dry chemical reagent tablet and the preparation method thereof of the invention have theadvantages of simple operation process, high stability, high good repeatability and high accuracy of detection results, quantitative detection, wide linear range, simple and convenient preparation process, easiness in operation, low risk and low pollution. According to the dry chemical reagent tablet and the preparation method of the dry chemical reagent tablet of the invention, the reagents do not need to be prepared, and samples can be directly added dropwise; the reagent tablet is dry and is free of moisture; the diffusion layer has a filtering function, so that hemolysis, bilirubin and ascorbic acid samples exert no significant interference on measurement results.