The purpose of our research was to evaluate the trypsin inhibitors content in five soybean genotypes obtained at IGPhPP. TI determination was performed according to the standard method, based on the ability of legume flour extracts to inhibit trypsin activity against the chromogenic substrate N-benzoyl-DL-arginine p-nitroanilide (BAPA), which was performed at MSU. As a result of the evaluation of the five soybean genotypes: Genap 54, L.2, Z3M10200, Onika and Z1M9250, where Genap 54 and L.2 which belong to the medium vegetation group, recorded a trypsin inhibitors content ranging from 12.31 mg/g, respectively 12.85 mg/g. These values indicate a moderate content of trypsin inhibitors. On the other hand, the genotypes Z3M10200, Onika and Z1M9250 belong to the semi-early vegetation group and recorded a higher content of trypsin inhibitors. More precisely, the values reported for these varieties were 14.06 mg/g, 15.06 mg/g, and 15.60 mg/g, respectively. These results are important from the perspective of using soybeans in food and the food industry. Although trypsin inhibitors can have an anti-nutritional effect by interfering with digestion activity, the moderate concentrations of inhibitors in Genap 54 and L.2 genotypes indicate that these varieties can be consumed properly and in reasonable amounts as part of a soy food. However, genotypes Z3M10200, Onika, and Z1M9250, which exhibited higher trypsin inhibitor contents, may require greater attention in terms of processing and consumption. By employing appropriate processing techniques such as soaking, boiling, or fermentation, the trypsin inhibitor content can be reduced, resulting in soy products that are more easily digestible and have increased nutritional value. In conclusion, our study has shown that the analyzed soybean genotypes exhibited variable trypsin inhibitor contents, and this information can be valuable in the development of processing strategies and techniques.