The dependence of electronic absorption spectra antimicrobial property on the substituent position was investigated using three Schiff bases derived from salicylaldehyde and isomeric aminobenzoic acids in three solvents of different polarities. The absorption maxima in all three solvents exhibited dependence on the position of substituent with the absorption maxima undergoing a red shift as solvent polarity increased. The in vitro antibacterial activity of the compounds against some clinically important bacteria namely Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Staphylococcus aureus (ATCC 25923), Enterococcus feacalis (ATCC 29212) was determined in N,N"-dimethylformamide and 1,4-dioxane using the agar dilution method. The results revealed that the ortho and meta substituted Schiff bases exhibited better antimicrobial activity in the non-polar solvent.