In this study, the chemical constituents and antioxidant property of holy basil (Ocimum sanctum Linn.) field-grown plant parts (leaves, stems, and inflorescence) were compared with those of respective callus cultures induced from each explant in in vitro. The callus cultures were successfully initiated on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) (1 mg/L) combined with different concentrations (0.1-0.5 mg/L) of kinetin as plant growth regulators. The distribution of phenolic compounds in these extracts was analyzed using reverse phase high-performance liquid chromatography with reference standards. Interestingly, rosmarinic acid (RA) was found to be the predominant phenolic acid in all callus extracts in comparison with field-grown plant parts. In this study, the antioxidant activity of the extracts was evaluated with six different in vitro antioxidant-testing systems. Their activities of scavenging superoxide anion radicals, 1,1-diphenyl-2-picrylhydrazyl radicals (DPPH), hydroxyl radicals, hydrogen peroxide, chelating ferrous iron, and ferric ion reducing potential were assessed. The antioxidant activity was increased in all testing systems with increasing amounts of extract. However, at the same concentration, the callus extracts exhibited higher antioxidant activity in all of the testing systems than the extract obtained from field-grown plant parts. The data obtained from this study suggested the possibility of the isolation of a high content of RA from in vitro callus cultures rather than field-grown plant organs of holy basil.