The seeds of Vaccaria hispanica (Mill.) Rauschert plant which spreads naturally in all regions of Turkey, have potential use in food and pharmaceutical industries. However, this plant, which is not well known, is not yet cultivated and used industrially. For sustainable industrial production, it is important to cultivate the seeds to obtain standard characteristic components such as superior starch granules, triterpenic saponins and cyclopeptides. The main objective of this study is to determine the content of the target components of the seeds obtained after adaptation to cultivation conditions among Vaccaria hispanica genotypes grown in nature. For this purpose, Vaccaria hispanica genotypes were collected, cultivated and analyzed regarding certain physical and chemical properties of seeds. The starch yield, segetalin A and saponin content of Vaccaria hispanica seeds were found in the range of 23.67-49.59 g/100 g dry seed, 0.17-0.65 g/100 g dry seed and 0.15-1.14 g/100 g dry seed, respectively. The thousand-grain weight, bulk density, moisture content, protein content, total phenolic and total fat content, and fatty acid composition were also determined. The seeds of Vaccaria hispanica genotypes were classified according to the analyzed parameters. All physical and chemical characteristics of the seeds subjected to principal component analyses and hierarchical cluster analyses which revealed that the total similarity values of the samples were determined to be 45.58% F1 and F2, 27.68% F1 and 17.90% F2; the samples were collected into three main groups.