In juice processing, ultrasound treatment has been tested as a potential alternative to conventional thermal methods to inactivate microorganisms and to enhance the nutritional status of juice. In this study, the impact of pasteurization and high-power ultrasound treatment on the quality of red grape juice was investigated in terms of the content of bioactive compounds such as phenolic compounds and l-ascorbic acid as well as regarding the microbiological and physicochemical properties. The grape juice was subjected to pasteurization (80 °C, 2 min) as well as to ultrasound treatment with an amplitude of 50 and 70% for 5 and 10 min. The results indicated the same level of total phenolic content for pasteurized and sonicated samples for 10 min with an amplitude of 70%, while the highest level of l-ascorbic acid was recorded for sonicated samples with an amplitude of 70% for 10 min. pH of sonicated samples decreased with amplitude and treatment time while total soluble solids and titratable acidity increased with amplitude and time. Moreover, the results indicated the usefulness of juice sonication to enhance the inactivation of microorganisms. Thus, the high-power ultrasound treatment might represent a viable technique to replace the conventional thermal treatment in grape juice processing.