Relevance. Milk contains a large number of biologically active compounds (including antioxidants) and is deservedly considered one of the main food products both in its original form and after technological processing.   Materials and methods. In this work, the measurements of the total amount of antioxidants (TOA) in milk samples of cows (from the «Ladozhskoye» breeding farm) at the peak of lactation were carried out by the amperometric method, and milk parameters during morning and evening milking were assessed by classical methods.   Results. As a result of the study, new data of TOA were obtained for 38 milk samples of Black-and-White breed cows of 5 groups depending on the lactation period: 1) 151–165 days; 2) 166–180 days; 3) 181–210 days; 4) 211–225 days; 5) 226–240 days, respectively. Moderate and strong correlations were obtained between TOA data and single milk yields (morning and evening milking), which are not only fundamentally new, but also important for assessing the total antioxidant activity in certain periods of lactation. Since the level of antioxidants significantly affects the shelf life, nutritional value, and many other parameters of milk, the authors suggest using the TOA level as an additional parameter in the overall assessment of milk quality. The data obtained are of great importance in assessing the physiological-biochemical status, including the state of the system of antioxidant defense of the body of cows.