Unveiling Phytochemicals and Antioxidant, Cytotoxic, Antithrombotic, Anti-Inflammatory, Analgesic, Hypoglycemic and Antidiarrheal Activities from the Leaves Extract of <i>Zanthoxylum rhetsa</i> (Roxb.) DC

Objectives This study aimed to investigate the chemico-pharmacological properties of the leaves methanolic extract (ME) of Zanthoxylum rhetsa (Roxb.) and its various solvent fractions, focusing on its antioxidant, cytotoxic, thrombolytic, anti-inflammatory, analgesic, hypoglycaemic, and antidiarrheal effects. Methods Phytochemicals in the ME were identified and characterized using gas chromatography and mass spectrometry (GC-MS). Antioxidant activity was measured via DPPH (2, 2-diphenyl-1-picrylhydrazyl) scavenging assay and measuring total phenolic content (TPC). Anti-inflammatory, thrombolytic, and cytotoxic effects were evaluated using membrane stabilization, clot lysis, and brine shrimp bioassays, respectively. In vivo evaluations of hypoglycaemic, antidiarrheal, and analgesic activities of the ME (200, 400 and 600 mg/kg bw) were conducted in mice model. Results A total of 65 phytoconstituents were identified in the ME, where 13-docosenamide (Z) (17.78%) and phytol (10.64%) were the most abundant. The aqueous soluble fraction (AQSF) fraction exhibited the highest TPC (181.875 mg GAE/gm of extractive) and antioxidant efficacy (IC 50 = 4.78 μg/mL). The AQSF also exhibited promising antithrombotic properties, showing over 88% clot lysis capacity. Furthermore, chloroform soluble fraction (CSF) displayed 93.2% haemolysis inhibition under heat-induced conditions, while the inhibition of haemolysis was 46.8% in hypotonic conditions. All the tested three doses showed dose-dependent statistically significant (p &lt; 0.05) analgesic and antidiarrheal efficacy, where only 200 and 400 mg/kg bw doses exhibited significant hypoglycaemic effects. In acute oral toxicity test, the ME exhibited median lethal dose greater than 4000 mg/kg bw. Conclusions Z. rhetsa leaf extractives demonstrated promising antioxidant, cytotoxic, antithrombotic, anti-inflammatory, analgesic and antidiarrheal effects, supporting the traditional uses of the plant species. Further extensive investigation, particularly on the AQSF fraction, is recommended to identify the responsible lead compounds.