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The objective of this research was to standardize the Green synthesis of silver nanoparticles by C.roseus and to evaluate them for their potential against Mycobacteral tuberculosis. Phytochemical study realized the presence flavonoids, saponins, alkaloids, cardiac glycosides, steroids and tannins. Colloidal silver nanoparicle was synthesized by one step green reduction at room temperature and characterized by UV and TEM. An effort made to find out the capping agent revealed that the TLC fraction with 0.7 Rf value showed the reduction of silver ions. GCMS analysis of TLC fraction showed the presence of nine different phytochemical which includes pentadecane and piperdine carboxylic acid as major constituent. The nanoparticle diameter is in the range of 38- 52 nm and stable for 30 days under room temperature. The bactericidal activity of synthesized nanoparticle was  24 ± 0.04 and 22 ± 0.62 mm respectively against M.smegmatis and M.tuberculosis. The in vitro hemolytic assessment of silver nanoparticles was found to be safe at maximum of 1-5 µg/mL and toxic between 15- 50 µg/ml. Further this data encourages that theses biologically synthesized nanoparticles were found to be less toxic and more effective against mycobacterial strains.

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