Thermal pasteurization is still one of the most effective methods for inactivating undesirable microorganisms in liquid foods. Pasteurization produces safer liquid foods with longer shelf-life. The batch thermal pasteurization process was standardized with different total heating time (fh) [(T1: Control-Un treated) (T2: 96°C, 540 s) (T3: 96°C, 720 s) (T4: 96°C, 900 s)]. Decimal reduction time ... [Show full abstract] (D96-value) of 1.5 min and Z-value of 10°C used to calculate maximum P-values. The effects of in-pack pasteurization on betalain pigment (Betacyanin and Betaxanthin), antioxidant (% of DPPH scavenging activity), CIE color (L* a* b* values), native micro flora and other physicochemical quality parameters were also evaluated during prolonged storage at ambient temperature (27-30 ± 2°C). The processing and storage of beetroot juice had a decisive impact and significant (p < 0.05) degradation in the betalain (betaxanthin and betacyanin) content, color and antioxidant activity during storage.