Comparative study of ultrasonic-assisted extracts of twelve traditional medicinal plants for in-vitro assessment of anticancer activity along with phytochemical and antioxidant potential

Abstract: Background: Pakistan has 6,000 plant species and several of these are known to have medicinal properties. However, dozens of plants are still scientifically unexplored or under-explored for their medicinal and therapeutic potential. Therefore, in the present study, twelve folk medicinal plants, including Plantago major, Cissampelos pareira, Justicia adhatoda, Myrsine africana, Rubus fruticosus, Santalum album, Solanum pseudocapsicum, Fumaria indica, Moringa oleifera, Quercus infectoria, Flacourtia jangomas and Acacia karroo, were chosen. Objectives: The study aimed to assess phytochemical content, antioxidant activity and anticancer potential. Methods: The extracts were prepared in 80% methanol under ultrasonic assistance. The antioxidant activity was measured using the DPPH method and the phytochemical content was evaluated following standard protocols. The cytotoxic potential was assessed using the MTT colorimetric assay against the HeLa cell line and multivariate regression analysis identified the common group (s) of bioactive metabolites among the studied plants. Results: The extraction yield was between 6.0% and 14.6%, depending on the plant material. Phytochemical analysis revealed varying levels of phenolics, flavonoids, tannins and alkaloids. Among all tested plants, M. africana exhibited the highest DPPH free radical-scavenging activity at 68.3 ± 1.2%, just below ascorbic acid (75.9 ± 1.5%), while J. adhatoda had the lowest (42.3 ± 2.5%). The MTT assay indicated that all plant extracts, except J. adhatoda, F. indica and F. jangomas, demonstrated promising anticancer effects, with M. africana showing the most potent activity (IC₅₀ = 37.7 ± 2.4 µg/mL). The two-way orthogonal PLS multivariate regression analysis identified phenolics and flavonoids as important classes of phytochemicals present in M. africana, S. album and M. oleifera, responsible for their potent antioxidant and cytotoxic properties. Conclusion: The results reveal that among the studied plants, M. africana exhibits the most significant antioxidant and cytotoxic potential. However, further research is recommended to isolate and identify the specific antioxidant and anticancer compounds.