Higher nitrogen fertilizer rates usually lead to an increase in photosynthetic pigments and can significantly affect not only the vegetative mass of plants, but also the content of photosynthetic pigments in plants, which is a key factor determining overall plant productivity. For this purpose, a three-factor field experiment was conducted in 2025 in the fields of the AgroITC Innovation and Research Center of the Agro Concern Group. The aim of this study was to investigate the influence of different nitrogen fertilization rates on the content of photosynthetic pigments in three winter wheat varieties (Factor A): 1) 'Chevignon', 2) 'LG Keramik', 3) 'Euforia'; Factor B—nitrogen fertilization rates: 1) 150 kg ha^-1, 2) 180 kg ha^-1; Factor C—nitrogen application time: 1) three times (BBCH 27–29; BBCH 29–30; BBCH 32–33); 2) four times (BBCH 27–29; BBCH 29–30; BBCH 32–33; BBCH 37–39). When fertilizing three times, the nitrogen rates were divided into N₆₀+N₆₀+N₃₀ and N₇₀+N₇₀+N₄₀, and when fertilizing four times, they were divided into N₅₀+N₅₀+N₂₀+N₃₀ and N₆₀+N₆₀+N₃₀+N₃₀. The photosynthetic pigments content was determined at BBCH 32-33 and BBCH 37-39 of winter wheat using the Holm–Wettstein methodology. The studies showed that the total amount of photosynthetic pigments and the mass of the aboveground part of the plants depended on the winter wheat variety and the level of nitrogen rates. The highest total amount of photosynthetic pigments was determined in the BBCH 32-33 stage in the winter wheat 'Chevignon', when the plants were fertilized with N₁₅₀, applied four times. However, the total amount of photosynthetic pigments in the BBCH 37-39 stage was the highest in 'Chevignon', when the plants were fertilized with N₁₅₀, applied three times.