Further investigation of material properties during the extrusion process is essential to achieve precise control of the quality of the extrudate. Whole oat flour was used to produce low moisture puffed samples by a twin-screw extruder. X-ray diffraction (XRD), Scanning electron microscopy (SEM), infrared spectroscopy (FTIR), thermal analysis, and rheological experiments were used to deeply characterize changes in the structure and cross-linking of oats in different extrusion zones. Results indicated that the melting region was the main region that changed oat starch, including the major transformation of oat starch crystal morphology and the significant decrease of enthalpy representing the starch pasting peak in the differential scanning calorimeter (DSC) pattern (p < 0.05). Moreover, the unstable structure of the protein increased in the barrel and then decreased significantly (p < 0.05) after being extruded through the die head. The viscosity of oats increased in the cooking zone but decreased after the melting zone. A transformation occurred from elastic-dominant behavior to viscoelastic-dominant behavior for oats in the melting zone and after being extruded. This study provides further theoretical support for the research of the change of materials during extrusion and the development of oat-based food.
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