Jia Wei San Huang Tang (JWSHT) alleviates the cold properties of the original formula, San Huang Tang, and holds promise for treating gastrointestinal ailments. The primary objective of this study was to validate the safety of the JWSHT in a rat model and position the experimental foundation for future comprehensive investigations into its pharmacological effects and safe clinical application. Guided by the "Methodology of Pharmacological Research on Traditional Chinese Medicine," JWSHT underwent oral toxicity assessments. In the acute study, 60 Kunming mice (half male, half female) were categorized into five groups, receiving gavage doses of 16.0, 12.8, 10.2, and 8.2g/kg JWSHT for 7 days. Symptoms and mortality were recorded, and LD50 was calculated. To ascertain the maximum tolerated dose, mice received 128g/kg of the product and were observed for 14 days. In the subacute rat study, 80 rats were divided into three treatment groups and one control, administered doses of 16g/kg/d, 8g/kg/d, and 4g/kg/d for 30 days. Subsequently, rats were euthanized, and diverse parameters were analyzed to evaluate JWSHT's subacute toxicity. The acute toxicity test revealed that the LD50 was greater than 5g/kg. No signs of toxicity were observed in mice when administered at the maximum dose. The results of the subacute toxicity test indicated that the hemoglobin (HGB) levels in the high-dose group and the mean platelet volume (MPV) in the low-dose group were significantly higher than those in the control group (P<0.05). The alanine aminotransferase (ALT) levels in the low-dose group were significantly lower than the control group (P<0.05). The relative spleen weight in the male high-dose group was significantly higher than that in the control group (P<0.05). Mild bleeding was observed in the kidneys of the high-dose group, while other parameters showed no significant difference compared to the control group (P>0.05). Therefore, it was concluded that under the conditions of this study, the administration of JWSHT was relatively safe.