Objective: To establish the chemical fingerprint of various secondary metabolites of Zanthoxylum rhetsa (Roxb.) DC, a medicinally important plant. Methods: Preliminary phytochemical screening for various secondary metabolites was carried out. HPTLC profiles of various individual secondary metabolites were done and profiles were developed for authentication. Result: The ethanolic extract of the fruit showed the presence of 8 Glycosides, 10 Flavonoids, 6 Essential Oils, 5 Anthraquinones, 9 bitter principles, 7 Coumarins and 8 Terpenoids. Conclusions: The development of such fingerprint for the fruits of Zanthoxylum rhetsa (Roxb.) DC is useful in differentiating the species from the adulterant and also act as biomarker for this plant in the Pharmaceutical industry.