OBJECTIVE: To investigate the effects of Qushi huayu formula on serum free fatty acid profile in non-alcoholic fatty liver disease (NAFLD) model rats. METHODS: A total of 30 SD male rats were divided into normal group, model group, Qushi huayu formula group, with 10 rats in each group according to random number table. Model group and Qushi huayu formula group were given high-lipid diet to establish NAFLD model; normal group was given common diet for 8 weeks. Since 5th week, Qushi huayu formula group was given Qushi huayu formula liquid 0.1 mL/kg intragastrically; normal group and model group were given constant volume of normal saline intragastrically for consecutive 4 weeks. 8 weeks later, the rats were sacrificed and liver tissue was collected. Histopathological changes of liver were observed by HE staining. Serum samples were collected and determined by GC/MS using menthyl salicylate as internal standard. The content of free fatty acid in serum was calculated by standard curve method. RESULTS: Compared with normal group, hepatocytes of rats in model group were necrotic, and fatty degeneration and vacuolar degeneration of hepatocytes appeared. The serum contents of saturated fatty acids as lauric acid, myristic acid, pentadecanoic acid, palmitic acid, heptadecanoic acid, stearic acid, arachidic acid, heneicosanic acid and docosanoic acid were increased significantly (P<0.05); the contents of unsaturated fatty acids as cis-10-heptadecenoic acid, oleic acid, α-linolenic acid, γ-linolenic acid, erucic acid, linoleic acid, cis-11-eicosenoic acid, arachidonic acid, cis-11, 14, 17-eicosatrienoic acid, cis-4, 7, 10, 13,16,19-docosahexenoic acid (DHA) were decreased significantly (P<0.05 or P<0.01). Compared with model group, histopathological changes of liver was relieved significantly in Qushi huayu formula group; serum contents of above saturated fatty acids were decreased significantly, while the contents of above unsaturated fatty acids were increased significantly (P<0.05). CONCLUSIONS: Qushi huayu formula can significantly improve histopathological changes of liver in high-lipid diet-induced NAFLD model rats, and can significantly improve the abnormal changes in the serum contents of saturated fatty acids and unsaturated fatty acids so that free fatty acid profile return to normal. It is suggested to supplement ω-3 polyunsaturated fatty acids or using medicine like Qushi huayu formula to intervene in fatty acid metabolism so as to effectively prevent and treat NAFLD.