The quality of potatoes intended for consumption or food processing depends primarily on their variety and chemical composition, which can be modified by growing conditions, place of cultivation, type of soil, amount of rainfall, as well as fertilization and plant protection products. This study aims to investigate the content of biologically active compounds in new varieties of potatoes with purple and yellow flesh, harvested in different cultivation locations. The content of the basic chemical composition and biologically active compounds in the raw material depended primarily on their variety, and to a lesser extent on the place of cultivation. Higher levels of kynurenic acid (KYNA), phenolic acids, and amino acids were found in potatoes of the new variety with purple-fleshed Provita compared to potatoes with yellow-fleshed Ismena. Potatoes with purple-flesh were characterized by over 40% lower amount of total glycoalcaloids (TGA). Most of the determined biologically active compounds, i.e. anthocyanins, phenolic acids, KYNA, and TGA varied depending on the place of origin of the potatoes, at the level of 40–66%. The biggest differences were found in the content of flavonols and amino acids, in the case of these substances the differences between potatoes from different places of cultivation ranged from 88% to 99%.