The objective of this work was to compare the effectiveness of three chemical families, namely, chlorogenic acids, fatty acids, and elements, for the discrimination of Arabica varieties (traditional versus modern introgressed lines) and potential terroirs within a given coffee-growing area. The experimental design included three Colombian locations in full combination with five (one traditional and four introgressed) Arabica varieties and two field replications. Chlorogenic acids, fatty acids, and elements were analyzed in coffee bean samples by HPLC, GC, and ICP-AES, respectively. Principal component analysis and discriminant analysis were carried out to compare the three methods. Although elements provided an excellent classification of the three locations studied, this chemical class was useless for Arabica variety discrimination. Chlorogenic acids gave satisfactory results, but fatty acids clearly offered the best results for the determination of both varieties and environments, with very high percentages of correct classification (79 and 90%, respectively).