BRINE SHRIMPS LETHALITY ASSAY OF THE EXTRACT PRODUCED BY ENDOPHYTIC FUNGI ISOLATED FROM THE ROOT OF Lowsonia inermis

Abstract

Abstract Endophytic fungi produced a secondary metabolites similar to their host plant; therefore it can be used as a source of producing active metabolites that can be used in drug discoveries. This work was aimed to isolate cytotoxic compounds from Endophytic fungi hosted in the root (tissue) of lawsonia inermis plant. The root of this plant was collected, prepared and screened for endophytic fungi. Aspergillus Niger was isolated, fermented on malt broth, extracted with ethyl acetate and partionioned using separating funnel. The crude ethyl acetate extract was tested for in vivo Brine shrimp lethality Assay of the extract and pure isolates. Cytotoxicity of the extract was evaluated in term of Lethal Concentation at 50% confident level (LC50). Ten shrimps were added into three replicates of each concentration 1000, 100, 10μg/ml respectively. After 24hours the number surviving shrimp were counted and LC50 was determine. The bioassay from the ethyl acecate fraction (HR-ASN) of the metabolite was obtained using column chromatography, which lead to the isolation of three pure compounds HR66, HR94, HR102. The result showed that the crude extract HR-ASN have LC50 of 3.15μg/ml, and that of the isolated compounds HR66, HR94, HR102 have the LC50 values of 4.094 μg/ml, 5.27 μg/ml, 2.297 μg/ml. HR102 revealed more potent activity oftoxicity with LC50 value of 2.295μg/ml. Therefore all the tested sample are toxic. It indicated that bioactive components are present and could be the source of pharmacological effect.

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