PHYTOCHEMICAL STUDIES AND EFFECT OF METHANOL LEAF EXTRACT OF LEPTADENIA HASTATA(PERS.) DECNE (ASCLEPIADACEAE) ON ACETIC ACID INDUCED WRITHES IN MICE AND VENOM OF ECHIS OCELLATUS
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ABSTRACT
Leptadeniahastata (Pers.) Decne (Asclepiadaceae) is widely African plant species used in traditional medicine as an anti-venom, antihypertensive, anti-diabetic, analgesic and also used for catarrh and skin disease. The powdered leaves of L. hastatawas extracted with methanol using maceration method and the resulting crude methanol extract (ME) was suspended in water and fractionated into hexane (HF), chloroform (CF), ethylacetate (EF) and n-Butanol (BF). Preliminary phytochemical screening of ME revealed the presence of carbohydrates, alkaloids, flavonoids, anthraquinones, saponins, tannins, glycosides, steroids and triterpenes while (HF) revealed the presence of steroids and triterpenes nucleus. HF was step-wisely eluted in a silica gel packed column to afford ten fractions H1-H10 and fraction H4 was selected for further purification using Preparative Thin Layer Chromatography, with the solvent system of hexane: ethyl acetate (8.5 : 2.5). This resulted in the isolation of a whitish crystalline compound M1 (βSitosterol). The structure of the compound was elucidated using chemical test and spectroscopic techniques (IR, 1D & 2D-NMR) and by comparison with reference spectral data. The LD50 of the methanol extract (ME) and the LD99 of the Echisocellatus venom were estimated to be greater than 5000 mg/kg and 2.2 mg/kg respectively. Anti-venom studies suggest that ME possess significant anti-venom activity against Echisocellatusvenom both In vitro and In vivo. Analgesic effect was evaluated using acetic acid induced writhing in mice. The results showed that ME (150 mg/kg) decreased writhing response with 84.6 % inhibition. However, the extract when compared to control showed statistically significant difference at (p˂0.05)
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