Vol. 7, Issue 2, Part B (2025)

Snakebite envenomation remains a significant cause of morbidity and mortality in tropical and subtropical regions, with conventional antivenom therapy facing limitations such as cost, hypersensitivity reactions, and limited effectiveness against local tissue damage. This study aimed to isolate and characterize a bioactive antivenom compound from the root-bark extract of Catunaregam nilotica, a medicinal plant widely used in African ethnomedicine for the treatment of snakebite. Dried root-bark of Catunaregam nilotica was extracted using absolute methanol and fractionated via successive solvent partitioning into n-hexane, ethyl acetate, and butanol fractions. The n-hexane fraction, which exhibited the highest inhibitory activity against phospholipase A₂ (PLA₂) activity was subjected to column chromatography to yield column chromatographic fractions (CPFs). These were screened in vitro for their inhibitory activity against snake venom metalloproteinases (SVMPs) and PLA₂ enzymes. The most potent chromatographic pooled fraction was characterized by gas chromatography-mass spectrometry (GC-MS), Fourier-transform infrared spectroscopy (FTIR), and UV-Visible (UV-Vis) spectroscopy techniques. Among the CPF18-b demonstrated the most potent inhibitory activity, significantly (p<0.05) reduced SVMP and PLA₂ activity to (1.53 ±0.20 ) and (2.99 ± 0.08, respectively. GC-MS analysis of CPF18-b revealed a predominant single peak corresponding to P-Menth-1-en-8-ol as the possible monoterpenoid compound. FTIR spectra exhibited prominent absorption bands indicating hydroxyl (-OH) stretching at ~3372 cm^-1 and C=C stretching at ~ 1630 cm^-1 while UV-Vis spectral analysis showed a characteristic absorption maximum at 210.5 nm supporting the presence of an unsaturated alcohol functional group which is consistent with a conjugated P-Menth-1-en-8-ol structure. This study provides the first evidence of linking p-Menth-1-en-8-ol to the antivenom efficacy of Catunaregam nilotica, thereby supporting its traditional use in the treatment of snakebite.