Grape seeds are a rich source of phenolic substances, which are known for their potent antioxidants and antimicrobial properties. This study aims to estimate the total phenolic content and antioxidant activity in the aqueous extract of red and white grape seeds. Additionally, the effect of adding red and white grape seeds in the form of powder or aqueous extract as preservative films by incorporating carboxymethyl cellulose (CMC) on the quality characteristics of beef burgers during freezing storage at -18±1°C for 90 days. The results showed that grape seeds contain phenolic substances at a high rate in addition to high antioxidant activity in both varieties. Free phenolic contents and the antioxidant activity were 557.83 - 411.70 mg GAE/100g and 66.37 - 72.71% in the aqueous extract of white and red grape seeds, respectively. Addition of grape seeds led to improving the characteristics of the beef burger during freezing storage. The values of pH, peroxide, and TBA decreased compared to the control sample. The same treatments caused an improvement in the microbial quality of beef burger; the total counts of bacteria, fungi, and yeasts were significantly decreased in the treated samples during storage compared with the control sample. In general, addition of grape seeds powder or their extracts as a natural source of antioxidants and antimicrobials instead of the synthetic antioxidants such as butylated hydroxytoluene (BHT) can improve the quality characteristics of beef burger and increase its acceptability among the consumers.