Identification of closely related plant species is not a trivial task, and is often difficult on the basis of morphological or anatomical features. To differentiate among analyzed species, a chromatographic fingerprint is usually constructed. This is not easy for very complex samples, however, especially those containing substances spanning a wide polarity range. In such circumstances more than one fingerprint is constructed, e.g. one for polar and the other for apolar constituents. In this paper a new method has been used for resolution of a mixture of 17 coumarins and flavonoids and, subsequently, construction of a fingerprint of closely related species belonging to the Peucedanum genus. This is the first time multidimensional planar chromatography has been applied for such purpose. Distinctive fingerprints are produced for every analyzed plant species, without the need to construct two fingerprints and without the application of more sophisticated multidimensional methods (e.g. hyphenation of HPLC and TLC, HPLC-MS, etc).