Abstract
The present research stated that 1,4-Bis(imidazolylazo)benzene (BIAB) has been prepared and spectrally characterized by the reaction of 1,4-diaminobenzene diazonium chloride salt with imidazole in an alkaline ethanol solution. Differing spectral techniques tested the compound preparation (BIAB) for elemental analysis 1H-NMR, UV-Vis, Infrared FT-IR, XRD, FE-SEM, thermal analysis (TGA-DTA) and C.H.N elemental analyses. The BIAB (biological ligand) was tested in-vitro for the various techniques for Staphylococcus aureus (Gram-positive) and Escherichia coli (gram-negative). The ligand BIAB's anti-fungal activity is also checked against two separate Aspergillus Niger and Candida albicans fungal species. Many of the examined fungal species were slightly active involved. The acute toxicity studies lethal doses (LD50) were calculated based on the Miller and Tainter determinations of LD50 in mice (Estimated Probity Units), findings of which indicate a 1020.23 mg/kg acute toxicity of LD50 when the mice were given intraperitoneally. The DPPH assay is used to assess a substance's ability to scavenge free radicals. The half maximal inhibitory concentration (IC50) is 224.17 µg/ml.

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