By cluster analysis, a comparative study was conducted for chemical components of 464 flue-cured tobacco samples with grades of C3F in tobacco growth regions of Shandong Province. The results showed that the average chemical components of flue-cured tobacco samples in Shandong were almost in normal range of high quality flue-cured tobacco, which indicated that the flue-cured tobacco in Shandong had appropriate content of chemical components, and had high quality. By systematical clustering method, the tobacco regions of Shandong were divided into five sub-regions of middle sugar-middle alkaloid- middle chlorine areas, lower sugar - high alkaloid - middle chlorine areas, high sugar-middle alkaloid- middle chlorine areas, high sugar-low alkaloid-middle chlorine areas, and high sugar-middle alkaloid-low chlorine areas. By using multivariate analysis of variance and Student-Newman-Keuls multiple-range test, the results showed that the difference among the locations were more significant.