Isolation Of Anticancer Drug Taxol Producing Endophytic Fungus Alternaria Alternata By Using HPLC And LC- MS

Abstract

Abstract Introduction/Objective In the exploration of an alternative source, optimum and numerous solvents with different percentage were exploited to extract Taxol from plant palm branche. The fungal endophytes producing secondary metabolites that are effective against the human infections have fascinated many researchers across the world. Among these, the exploration for novel cancer therapeutic agents is of countless reputation due to upsurge in the number of cancer deaths, high worth of the drugs and the side effects concomitant with cancer treatments. Accordingly, in the current study, an endophytic fungal strain was isolated from its host plant palm branche and identified as Alternaria Alternata based on its morphological features and internal transcribed spacer sequence analysis. Methods Firstly, palm branche were collected for the isolation of endophytic fungi. The samples were placed on PDA plates and after 72 h, the mycelium produced were isolated and sub-cultured. Secondly, further analysis for the presence of Taxol by using chromatographic and spectrometric techniques. Illustratively, ethanol or acetone and 50% and 20% of ethanol. Then, the residues were dissolved in 5 ml of methanol and filtered. By using the internal standard, the resulting solution was subjected to the HPLC to determine the extraction efficiencies of tested solvents. Finally, filtrate was also subjected to the LC-MS – ESI using water/acetonitrile (10/90) as mobile phase to identify the authenticity of Taxol Results HPLC analysis revealed that Taxol could be quantified by UV detection using standard curve. Furthermore, the standard curve covering the concentration was linear (r2= 0.9992) and CV% ranged from 0.52 to 15.36. The peaks of fraction 5 obtained by HPLC analysis were quite similar to that of standard taxol confirming the presence of taxol. A parent ion peak of m/z 851.85 was observed in the LC–ESI–MS spectrum which was similar to paclitaxel with reported m/z of 854 [M+H] + ion. A. Alternata produced about 113.193 mg/L taxol as quantified through HPLC Conclusion we are recording for the first time the production of taxol by an endophytic fungus Alternaria Alternata. In a final word, present study results advocate that the endophytic fungus A.Alternata aids as a potential source for the production of taxol isolated from Palm branche plant