Senna italica as a medicinal plant has wide usage in traditional medicine, particularly by communities in different districts of the Limpopo province, South Africa. The medicinal value of plants parts is dependent on the nature of phytochemicals they possess. The aim of the study was to obtain and compare the TLC and UV-Vis spectral profiles of S. italica collected from four districts in Limpopo province of South Africa. The leaves of S. italica were collected from four different districts (Capricorn, Sekhukhune, Vhembe and Waterburg districts) in the Limpopo province of South Africa. The dried, ground leaves were extracted with solvents of different polarities. Constituents of the different extracts were resolved on TLC plates using different solvent systems and the Rf values of resolved compound bands noted and compared between collection districts. The UV-Vis spectra of different extracts were obtained and wavelengths of compound peak signals noted and compared between collection districts. The obtained results showed in some instances a disparity on the TLC and UV-Vis spectral profiles of the leaf extracts of S. italica between collection districts. This was evidenced by the presence of compound bands at Rf values and/or presence of compound peak signals at wavelengths in other locations which were absent in other locations of occurrence within extracts obtained by same solvent. Thus, it could be concluded that the phytochemical composition of the leaves of S. italica is affected by geographical location of occurrence within Limpopo province (South Africa).