In this study, we developed a liver tissue metabolomic method based on gas chromatography-mass spectrometry (GC-MS) to evaluate the effect of acute paraquat poisoning on rats. The acute paraquat poisoning group rats were given 36 mg/kg of paraquat by intragastric administration, control group were given saline by intragastric administration. Liver tissue samples were collected from the rats from the acute paraquat poisoning group and control group. Pattern recognition analysis, including both principal component analysis (PCA) and partial least squares-discriminate analysis (PLS-DA) revealed that acute paraquat poisoning induced liver tissue metabolic perturbations. Compared to the control group, the level of (Z,Z)-9,12-octadecadienoic acid, D-gluconic acid, oleic acid, 4,7,10,13,16,19-docosahexaenoic acid, 5,8,11,14,17-eicosapentaenoic acid, d-glucose, phthalic acid in liver tissue of acute paraquat poisoning group increased, while the level of d-mannose, heptadecanoic acid, D-glucuronic acid decreased. In conclusion, liver tissue metabolomic method based on GC-MS may be useful to elucidate acute paraquat poisoning through the exploration of biomarkers. According to the pathological changes of liver tissue at difference dosage, paraquat is hepatotoxic.