Increasing consumer awareness regarding the health benefits of different nutrients in food has led to the requirement of assessing the effect of food processing approaches on the quality attributes. The present work focuses on understanding the effects of ultrasound (US) processing, mild heat pasteurization (65˚C for 15 min), thermal pasteurization (80˚C for 15 min) and their combination on physicochemical, microbiological properties and nutritional quality of pineapple juice through 60 days of storage at room temperature. Ultrasound treatment showed significantly lower browning degree. Ultrasound followed by ultrasound combined with mild heat pasteurization (UMP) treatments was effective in retaining the total phenolic content of pineapple juice as compared to the thermal treatment or the untreated juice sample at room temperature during 60 days of storage. Thermal pasteurization (TP) followed by ultrasound combined with mild heat pasteurization (UMP) and ultrasound (US) treatment, in increasing order, was found to be effective in delaying microbial growth in pineapple juice. This study demonstrates that ultrasound combined with mild heat pasteurization treatments could be able to effectively inactivate the microorganisms and pectin methylesterase in pineapple juice whilst preserving relatively high amount of phenols.