Alkyl Chain Length Governs Structure, Conformation and Antimicrobial Activity in Poly(alkylene biguanide)

Poly(hexamethylene biguanide) (PHMB) is a polycationic antimicrobial polymer exhibiting broad-spectrum activity against bacteria, fungi, and viruses, and is widely used in medical settings for infection prevention and control. However, the relationship between chemical structure and antimicrobial activity remains unclear. In this study, we synthesised and characterised a series of polymeric biguanides with systematically varied alkyl chain lengths to examine the effects of structural variation on physicochemical properties and antimicrobial activity. H NMR spectroscopy and FTIR confirmed successful polymerisation. Solubility measurements revealed a progressive decrease in aqueous solubility with increasing alkyl chain length, consistent with increased hydrophobicity. Dynamic light scattering indicated reversible folding and unfolding of polymer chains in aqueous solution, with stabilisation at higher concentrations. Diffusion-ordered spectroscopy was used to calculate hydrodynamic diameters and polydispersity indices. Antimicrobial assays against Staphylococcus aureus and Pseudomonas aeruginosa showed that polymers containing heptamethylene and octamethylene chains exhibited the highest antibacterial activity, whereas tetramethylene- and pentamethylene-containing polymers showed greater fungicidal activity against Candida albicans. Highly hydrophobic polymers showed increased aggregation, resulting in reduced antimicrobial efficacy. Overall, these results indicate that both charge density and alkyl chain length are key determinants of antimicrobial activity. This polymeric biguanide series provides a platform for further investigation of structure–activity relationships and mechanisms of action against pathogenic microorganisms and their biofilms.