Abstract: At present, starch and gluten protein is usually produced by rear road flour at home and abroad for the consideration of cost, although the yield is high, the quality is poor. Based on this status quo, this study compared and analyzed the quality and rheological properties of front, middle, and rear road flour in wheat flour making process by using farinograph, extensometer and rheometer, so as to explore the specific differences among the three basic powders. Among them, the farinograph can reflect the kneading resistance of the dough, and intuitively reflect the stability time and weakening degree of the dough. It can be used to evaluate the gluten strength of dough and determine the quality and application of wheat flour. The extensometer can directly reflect the stretching resistance and elongation of the dough. Therefore, it can be used to evaluate the elasticity of the dough and the strength of the protein in the dough. The rotary rheometer can provide the viscoelastic properties of the polymer network with different oscillation frequencies. Therefore, it can be used to reflect the elasticity and viscosity of dough and gluten. The rapid viscosity analyzer is an effective tool for analyzing the α-amylase activity and starch gelatinization characteristics of cereals and their products. It is beneficial for the industry to select specific flour mill streams mixing, so as to improve the quality of rear road flour. Previous studies have reported that ash content, protein content, dry gluten, wet gluten, falling value, gelatinization temperature, water absorption, and elongation of rear road flour in almost every wheat are higher than the front road flour. The whiteness, gluten index, peak viscosity, minimum viscosity, attenuation value, final viscosity, retrogradation value, peak time, stretching resistance, and maximum stretching resistance gradually decrease from the front to the rear flour, all of which showed obvious differences and regularities. The formation time of rear road flour dough is higher than that of front road flour, and there is no uniform regularity in the change of stability time, weakening degree and flour quality index. The content of damaged starch and water-soluble matter both decreased first and then increased. The rear road flour dough has high gas production capacity, high fermentation stability, and poor air retention. The middle flour dough has the highest expansion and the best air retention. Conclusion:There are same regular changes in the physicochemical properties of front, middle, and rear road flour during the same process of different wheat flour milling and rheological changes of the formed dough except for the stabilization time, weakening degree, and flour quality index. This study not only provides a reference for the system powder blending in the later stage, but also provides a theoretical guidance for improving the quality of starch and gluten protein produced by industry and the selection of raw flour.