Rana Baqa Muhammad1*, Tanveer Hussain1*, Muhammad Rafay1, Amjad Saeed1, Hassan Mehmood2, Aziza Huseynova3
1Institute of Forest Sciences, Faculty of Agriculture & Environment, The Islamia University of Bahawalpur, Bahawalpur-63100, Pakistan
2Institure of Agro-Industry and Environment, Faculty of Agriculture and Environment, The Islamia University of Bahawalpur-63100, Pakistan
3Department of Basic Medical, Nakhchivan State University, Baku, Azerbaijan
*Corresponding author’s email: ranbaqa02@gmail.com; dr.tanveer@iub.edu.pk
Received: 20 June 2026 / Revised: 19 September 2026 / Accepted: 07 October 2026 / Published Online: 12 October 2026
Abstract
The rising global problem of antimicrobial resistance and chronic diseases requires the discovery of novel therapeutic agents from natural sources. Medicinal plants possessing diverse bioactive compounds offer sustainable alternatives to synthetic pharmaceuticals. This study evaluated the phytochemical composition and biological activities of Portulaca oleracea L. collected from two distinct agro-climatic zones (Yazman and Kahror Pakka) in Southern Punjab, Pakistan during February 2024. The plant parts (leaves, stems and roots) were extracted through cold maceration using methanol. Proximate composition, quantitative phytochemicals (total phenolic, tannin, flavonoid and saponin contents), antioxidant activity (2,2-diphenyl-1-picrylhydrazyl, DPPH, assay), antibacterial efficacy against four pathogenic strains (Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae and Klebsiella pneumoniae), antifungal activity against four Aspergillus species and pharmacological potentials (in vitro cytotoxicity, anti-inflammatory and antipyretic) were assessed using the DPPH radical-scavenging assay (antioxidant), agar-well diffusion (antibacterial and antifungal), the MTT assay on murine 3T3 fibroblasts (cytotoxicity), zymosan-induced ROS chemiluminescence (anti-inflammatory) and an E. coli-induced pyrexia rabbit model (antipyretic). Three independent biological replicates were sampled per location and plant part (n = 3), each analyzed in analytical triplicate and the resulting plant means were subjected to two-way factorial analysis of variance (ANOVA) testing location, plant part and their interaction, followed by Tukey’s honestly significant difference (HSD) test (p ≤ 0.05). Kahror Pakka samples exhibited significantly higher crude protein (56.32-64.32%), moisture (6.49-7.21%) and ash (14.19-18.41%) contents, whereas Yazman specimens showed elevated crude fiber (20.43-56.23%) and crude fat (7.21-22.99%). Leaf extracts from Kahror Pakka demonstrated superior antioxidant activity (75.73% DPPH inhibition), total phenolic content (35.31 mg GAE g‑1), total flavonoid content (23.11 mg QE g‑1), maximum antibacterial zones of inhibition (11.57-14.57 mm) and enhanced pharmacological activities, including in vitro cytotoxicity toward 3T3 fibroblasts (59.40%), anti-inflammatory (61.40%) and antipyretic (34.83%) effects. Pearson correlation analysis revealed highly significant positive relationships (p ≤ 0.001) between secondary-metabolite contents (total phenolics, flavonoids, tannins and saponins) and the antioxidant, antimicrobial and pharmacological activities. Because only two locations were compared and location is inseparable from the accompanying edaphic and climatic differences, these site contrasts are descriptive and cannot attribute the observed variation to any single agro-climatic factor; within that constraint, leaves consistently showed the highest activity of the three organs at both sites. Portulaca oleracea from Kahror Pakka therefore warrants further investigation as a candidate source of antioxidant and antimicrobial constituents.
Keywords: Phytochemicals, Antioxidant, Antimicrobial, Pharmacological, Purslane, Methanol, Therapeutics
