Traditional medicine makes heavy use of the antioxidant and therapeutic qualities of the medicinal herb Boerhaavia diffusa. Using mass spectrometry, this study aims to identify and characterise the antioxidant components found in mother tincture of Boerhaavia diffusa. Subsequently, in vitro tests were conducted to validate the compounds' antioxidant ability. To find bioactive compounds, the study used mass spectrometry (MS). Roughly 6,000 compounds were found in the mass spectrometric analysis, with 99 of those molecules being particularly noteworthy for their strong antioxidant properties. The molecular weights and mass-to-charge ratios (m/z) were used for the classification of the compounds. Significant antioxidant activities were exhibited by prominent components such as ascorbic acid, alpha-tocopherol, beta-carotene, curcumin, and chlorogenic acid. Boerhaavia diffusa's ability to reduce oxidative stress is supported by its presence of flavonoids, carotenoids, and phenolic acids, which contribute to its therapeutic effect. The mother tincture's entire therapeutic efficacy is enhanced by the range of antioxidant components, which indicate a synergistic effect. Researchers found that the mother tincture of Boerhaavia diffusa contains bioactive antioxidant compounds that could help prevent or manage diseases caused by oxidative stress. To explore its therapeutic properties across different potencies and formulations for extended medicinal purposes, additional research is needed.