Abstract
Phytochemicals with protein binding properties, active against snake envenomation such as flavonoids, polyphenols, saponins, tannins, terpenoids, xanthenes, quinonoids, steroids and alkaloids bind to toxic venom proteins thereby inactivating them. This research was aimed at isolating antisnake venom agents from Azadirachta indica leaf extracts using activity-guided isolation protocols. A. indica leaf was collected, authenticated and extracted using methanol followed by solvent-fractionation using hexane and ethylacetate. These fractions were further separated using column and thin-layer chromatography. The pooled chromatographic fractions were screened for the antivenom activity using Naja nigricollis venom phospholipase A2 (NVPLA2) inhibition assay and albino rat models. Polyvalent serum-based antivenin was used as standard. The column chromatography of the A. indica leaf hexane and ethylacetate extracts yielded sixty (60) and sixty nine (69) fractions. Based on similarities on the TLC profiles, fractions were pooled. Eleven (11) pooled hexane fractions (PHFs) and fourteen (14) pooled ethylacetate fractions (PEFs) were obtained. After antivenom activity screening, pooled hexane fraction 3 (PHF3) and pooled ethylacetate fraction 1B (PEF1B) from the hexane and ethylacetate fractions respectively protected envenomed albino rats from death at a dose of 20 mg/kg b.w. against 0.8 mg/kg b.w. of N. nigricollis venom. Hence, these findings suggest that the isolated active compounds can serve as leads for the development of safe, readily available and affordable plant-based antivenoms.
Highlights
Snakebite is a neglected public health problem in most of the countries in Africa
The pooled chromatographic fractions were screened for the antivenom activity using Naja nigricollis venom phospholipase A2 (NVPLA2) inhibition assay and albino rat models
These plants are reputed to be capable of neutralizing the action of snake venom, and their antivenom activity has been related to certain chemical compounds identified in those plants [27]
Summary
Rural populations are frequent victims as they go about their daily food production and animal rearing activities and as they reside in the comfort of their homes [1]. Many of these snakebite cases go unreported and do not appear in official epidemiological statistics. Production of antivenins is a painstaking, resource-intensive and time-consuming process [2]. Another important issue with the conventional antivenins is the knowledge of which species of snakes inflicted the bite. Antivenins do not neutralize the local tissue damage [4]
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