Objective To create an optimal method for establishment of a rat model of constipation-predominant irritable bowel syndrome (IBS-C), and evaluate the traditional Chinese medicine (TCM) syndrome in this generated IBS-C model. Methods One hundred and thirty-three SPF neonatal rats of 20 litters were used in this study. Using maternal separation (MS), ice water stimulation, water limitation, MS combined with ice water stimulation, and MS combined with water limitation to establish rat constipation model. The Chinese medicine Xiao-Yao powder was administered to the rat models. To observe the general behavior, body weight and daily food-intake, to measure the defecation numbers and water content of the feces within 4 hours, and the types of feces were assessed by Bristol stool scale. Colorectal distention test was used to assess the intestinal sensitivity, and histological examination using haemotoxylin staining was performed to observe the alterations of intestinal tissues. Results MS combined with ice water stimulation was the best method to induce the rat models with decreased defecation number and decreased water content of feces. There were no obvious histological alterations in the intestinal tissue. The scores by Bristol stool scale indicated that the model was an IBS-C model with visceral hypersensitivity, and the results of Xiaoyao powder administration indicated that this IBS-C model met the requirement of clinical characteristics of liver-stagnation and spleen-deficiency syndrome. Conclusions Maternal separation combined with ice water stimulation is a good method to successfully establish an IBS-C rat model with liver-stagnation and spleen-deficiency, highly simulating the clinical symptoms of this syndrome.