Hexadecylpyridinium bromide is a cationic surfactant capable of bactericidal activity and has been used for ion exchange. As a bactericidal agent, it reduces the surface tension between pathogenic membranes and their environments. This interaction disrupts membrane integrity and osmoregulation, leading to cell lysis.

Hexadecylpyridinium bromide has also been shown to quickly and efficiently remove metal ions from solutions. The compound removes cations from solution through a cation/surfactant complex which precipitates from an aqueous phase into an organic phase and can subsequently be extracted. Hexadecylpyridinium bromide is an effective ion exchange agent for extracting DNA and proteins.