The preparation of polyelectrolytes based on crosslinked poly(vinyl alcohol) (PVA) and chitosan (CS) was considered as a feasible alternative to develop highly functionalised, cost-effective and eco-friendly membranes for proton exchange fuel cell technologies. CS/PVA-based membranes were combined with sulfosuccinic acid (SSA) as crosslinking and sulfonating agent, and glycerol (GL) to promote flexibility and favour their manageability. The chemical structure, the thermo-oxidative behaviour, the ethanol uptake, the electric, the proton conductivity, and the performance in direct ethanol fuel cell (DEFC) were assessed. In general, all the CS/PVA-based polyelectrolytes showed a synergetic increase of thermo-oxidative stability, appropriate absorption and diffusion of ethanol and good proton conductivity, suitable for the typical service conditions of fuel cells. The GL in the membranes reacted with SSA, reduced the ethanol absorption, the diffusion coefficient and the proton conductivity, but acted as a plasticiser that increased the ductile manageability of the polyelectrolytes to be mounted on the membrane-electrode assembly (MEA). Higher presence of CS and higher proportion of GL in the polyelectrolyte, improved the material performance in the DEFC. In particular, the crosslinked polyelectrolyte 40CS/PVA/SSA/20GL with a 40%wt. of CS referred to PVA, and a 20%wt. of GL referred to CS, showed a suitable behaviour in the DEFC test, with a maximum value of power density of 746 mW.cm(-2).