PHYTOCHEMICAL, ANTIOXIDANT AND ANTISNAKE VENOM STUDIES OF THE METHANOL EXTRACT OF THE ROOT BARK OF PHYLLANTHUS MUELLERIANUS (KUNTZE) EXCELL
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ABSTRACT
Phyllanthus muellerianus is distributed in secondary forests from Guinea-Bissau, Mali, West Cameroons and other parts of tropical Africa. It is a shrub or climber deciduous and occasionally arborescent. The root bark of this plant was extracted by maceration using methanol. The preliminary phytochemical screening of the methanol extract indicates the presence of terpenoids, flavonoids, tannins, saponins, alkaloids and carbohydrates. The methanol extract was then dissolved in water and partitioned with nhexane, ethyl acetate and n-butanol. The partitioned n-butanol fraction was subjected to column chromatography, gel filtration chromatography and then preparative thin layer chromatography to yield a brown pentasaccharide PA - α-D-Glcp-(2 6)-[ α-D-Fruf(1 2)]α-D-Fruf-(1 4)β-D-Glcp-(2 2)β-D-Fruf and sucrose PX - α-D-Glcp-(1 2)α-D-Fruf which is whitish in colour. The structures of the compounds were resolved using UV, IR, 1D- and 2D-NMR spectral results. n-Butanol was also added to the dry methanol extract. The n-butanol fraction of the dry crude extract was also subjected to gel filtration chromatography and preparative thin layer chromatography to give a brown amino alcohol glycoside CP1 – (2S, 3S, 4S, 5R)-2-((E)-3,10-diaminoicos-5-enyloxy)- tetrahydro-6-(hydroxymethyl)-2H-pyran-3,4,5-triol. The LD50 and LD99 in mice of the methanol extract and the venom of Naja nigricollis were determined to be 400mg/kg and 9.55mg/kg using Lorke’s method and probit analysis respectively. Different doses of the methanol extract and the LD99 of the venom were used for in vitro and in vivo detoxification studies in mice. The extract exhibited 67% protection against lethality at an administered dose of 20mg/kg, 83% at 40mg/kg and 33% at 80mg/kg of the extract in vitro while there was 50% protection against lethality at an administered dose of 10mg/kg in vivo. The isolated pentasaccharide exhibited a free radical scavenging activity comparable to that of vitamin C at 95% confidence level using DPPH as the viii standard free radical and methanol as solvent. Conclusively, the root bark of the plant is rich in saccharides and the ethnomedicinal claim as to the antivenin properties of the root bark of Phyllanthus muellerianus has been scientifically validated.
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