eng Oriental Scientific Publishing Company Biomedical and Pharmacology Journal 0974-6242 2022-06-30 15 2 1115 1126 10.13005/bpj/2447 44523 article Maysaa A.M. Alhar 1 Horticulture Department, Faculty of Agriculture, University of Kufa, Iraq. This study was conducted to create a new reagent from Anil-Azo compounds and study their biological impacts on two types of bacteria. For the first step, four reagents had been created by reaction of p-phenyl diamine in acidic form with 2-formyl-4-methyl phenol in neutral medium to create reagent {1}, which used to produce reagent {2} by reacting it with amino benzothiazole over four hours in the presence of glacial CH₃COOH. The reagent {1} was also used to form reagent {3} by reacting it with amino imidazole over two hours. Finally, reagent {3} had been generated by reacting reagent {1} with naphthyl amine (0.2 mol) over four hours in the presence of glacial CH₃COOH. The UV-visible spectrum was showed that a new ligand was created between 190-600 nm in reagent {2}, {3} and {4} while reagent {1} was appeared in 519-600 nm area. FTIR spectrum showed that many new coordinate bonds had been formed in different locations. Also, the chromatographic separation study showed that reagent {4} was separated faster than other reagents. Study of compounds stability showed that all reagents were stable in methanol, ethanol, DMSO and DMF. Study of chemical-physical peripteries showed that percentage of reagents’ yield ranged between 80-70%. The assessment of the formulated reagents against various kinds of bacteria was carried out using a medium (agar) via numerous processes. Microbial inhibition was tested at three concentrations: 30, 50 and 70 micrograms, with a blank solvent (DMSO), for bacteria Staphylococcus aureus, E. coli and Streptococcus pneumonia with an incubation period of 24 hours at 37℃. The results of biological impacts showed that reagent {2} showed more inhibition Staphylococcus aureus and Streptococcus pneumonia. https://biomedpharmajournal.org/vol15no2/a-study-of-the-spectral-chromatographic-and-solubility-characteristics-of-new-analytical-reagents-from-anil-azo-and-their-biological-effects-on-bacteria/ Analytical Antimicrobial Bacteria  Reagent