Objective To investigate the metabolite features of chronic pancreatitis in rats in vitro by high resolution magic angle spinning nuclear magnetic resonance spectroscopy. Methods A total of 30 Spraque-Dawley rats were randomized into experimental group (n =20) and control group (n = 10). All the animals in experimental group were intravenously injected with 8 mg/kg body weight DBTC, and the animals in the control group received same amount of solvent. Pancreatic tissues were examined by histology and Van Gieson staining. Metabolic changes of chronic pancreatitis in vitro in rats were studied by high resolution magic angle spinning nuclear magnetic resonance spectroscopy. Results 60 days after DBTC application, the pancreatic tissue was characterized by an extended interstitial fibrosis with infiltrating mononuclear cells. Compared with the control group, the signal intensities of phosphocholine (Pc) and glycerophosphocholine (Gpc), taurine (Tau), lactate (Lac) of chronic pancreatitis group increased. Oppositely, the signal intensities of betine (Bet), glutamic acid ( Glu ), alanine (Ala), ileucine (He), leucine ( Leu ) and valine (Val) decreased. The signal intensities of acetic acid (Ace) and choline (Cho) were not changed. Conclusions There were obvious metabolic features of chronic pancreatitis in rats, and it is helpful for the application of magnetic resonance spectroscopy in chronic pancreatitis in human studies.