The aim of this study was to evaluate winter wheat grain qualitative and yield parameters after the application of different organic and mineral fertilizers at two sites in the Czech Republic (S1 – Lukavec, S2 – Prague-Suchdol). For the purpose of this paper grain yield (GY, t ha−1), thousand kernel weight (TKW, g), bulk density (BD, g l−1), grain crude protein content (CP, %), and falling number (FN, s) were determined with regard to the CSN 461100-2 (2001) standard for food wheat. Significantly highest values of GY and BD at both sites and of CP at S2 were observed after the use of nitrogen in mineral form. At S1, significantly highest values of FN were obtained after the use of mineral fertilizers in combination with straw. The required limit of BD for food wheat was met for all fertilization treatments, while that of CP only at S2 by treatments with mineral nitrogen fertilizers. At S2, a strong correlation between all evaluated traits was registered. Low correlation was found at S1 between TKW and other evaluated traits.