<div><p><em>We report herein synthesis, characterization and antimicrobial activity of four Shiff bases derived from hexamethylenediamine. Substitution of a nitro group on each aromatic ring in ortho, meta or para positions of N,N'-bis(phenylmethyl)hexane-1,6-diimineallowed to have a homogeneous series of positional isomers. These four symmetric diimine Schiff bases were characterized by conventional spectrometry methods (NMR, IR, MS), then tested against Gram-positive and Gram-negative bacterial strains. Among them, compounds <strong>1b</strong>, <strong>1c</strong>, <strong>1d</strong> were found to be active against bacterial strain Staphylococcus aureus CIP with MIC value of 375</em><em>μ</em><em>g/ ml, 187.5</em><em>μ</em><em>g /ml and 375</em><em>μ</em><em>g /ml respectively. Candida Albicans fungal strain showed resistance to all synthesized Schiff base compounds, butin the other hand, Candida glabrata has been sensitive to all compounds with MIC of 1500</em><em>μ</em><em>g/ ml and one more time except <strong>1a</strong>.</em></p></div>
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