A series of 4-amino-1-alkyl pyridinium bromide salts (1–10) were synthesized and antimicrobial activity was evaluated by disc diffusion as well as broth macro-dilution method. 4-Amino-1-hexadecylpyridinium bromide (10) which showed good inhibition against Candida albicans and C. glabrata also displayed good germ tube inhibition against C. albicans. Compound 8 shows very low haemolytic activity against human red blood cells. Compound 10 exhibits excellent antibacterial activity against Escherichia coli and Staphylococcus aureus. Inhibition of phospholipase enzyme secretion was also observed against compound 8 to find the possible mode of action.