Abstract: In order to investigate the physicochemical characteristics of different segments of cinnabar flue-cured tobacco leaves and to provide a reference for the precise classification of the tobacco in threshing and redrying procedures, the middle leaves were sampled from Yunnan Province, and each single leaf was evenly divided into 25 segments along the longitudinal direction after removing the petiole. The physicochemical characteristics of each segment were determined by a spectral spectrophotometer, thickness gauge, horizontal tension meter, and continuous flow analyzer. The variations of the properties from the different segments were analyzed by regression. The appropriate segmentation number and cutting ratio of the tobacco leaves were determined by cluster analysis, and their rationality was verified by chemical composition and sensory quality evaluation. The results showed that: 1) The physicochemical characteristics of cinnabar flue-cured tobacco leaf presented a differential distribution and regular change trends from the different segments and had a good linear relationship (R² > 0.818 2). 2) The principal component analysis and clustering results suggested that there were certain differences and similarities in the physicochemical characteristic indices based on leaf segments, which was suitably divided into three segments with a cutting ratio of 32.31%∶55.38%∶12.31% for the base∶the middle∶the top. 3) There were significant differences in chemical components such as total sugar, plastid pigment, total polyphenol and volatile aroma components among these segments (P < 0.05). Glutinous rice, hay-like and burnt-sweet aroma notes in the middle segment were prominent with rich aroma and smoke quantity, mild and smooth notes, less offensive odor and irritation, and better overall sensory quality. Furthermore, it could be concluded that threshing and redrying different segments of cinnabar tobacco leaves separately was more conducive to increasing the utilization value of tobacco leaves and their contribution to cigarette products.