This study investigates hydrogen production from biomass feedstocks (glucose, starch, lignin, and cellulose) using a 100 mL h-type proton exchange membrane electrolysis cell. Biomass electrolysis is a promising process for hydrogen production, despite its low technology readiness level, due to its advantages such as lower-temperature conditions, minimal energy consumption, potential for high-volume hydrogen production, and smaller carbon footprint compared to thermochemical processes. A Lewis acid (FeCl₃) was used as a charge carrier and redox medium to aid in biomass depolymerisation/oxidation. A comprehensive analysis was conducted, measuring hydrogen and CO₂ emission volumes and performing electrochemical analysis (linear sweep voltammetry and chronoamperometry) to understand the process. The highest hydrogen volume was 12.1 mL, produced by FeCl₃-mediated electrolysis of glucose at ambient temperature, which was up to two times higher than starch, lignin, and cellulose at 1.20 V. Glucose also showed a maximum power-to-hydrogen yield ratio of 30.99 kWh/kg. The results showed that hydrogen can be produced from biomass feedstock at ambient temperature using FeCl₃, with higher yield and lower electricity consumption compared to water electrolysis. The study also found that the hydrogen content of the feedstock is a key factor in hydrogen production. The maximum hydrogen yields varied from 6.3 to 12.1 mL, with glucose showing the highest yield. The power-to-hydrogen yield ratio at ambient conditions ranged from 30.99 to 33.83 kWh/kg, which is more energy-efficient than conventional water electrolysis (55–58 kWh/kg). CO₂ emissions were also investigated, showing low emissions due to a maximum carbon consumption rate of around 13%. The study concludes that biomass electrolysis using FeCl₃ is a promising method for hydrogen production, with potential for cost-effective and sustainable hydrogen generation. The results suggest that biomass electrolysis can be operated at ambient temperature, with a higher yield and lower electricity consumption compared to water electrolysis. The study also highlights the importance of feedstock selection based on hydrogen content and biomass structure. The findings contribute to the development of low-carbon-emission hydrogen production from biomass resources.This study investigates hydrogen production from biomass feedstocks (glucose, starch, lignin, and cellulose) using a 100 mL h-type proton exchange membrane electrolysis cell. Biomass electrolysis is a promising process for hydrogen production, despite its low technology readiness level, due to its advantages such as lower-temperature conditions, minimal energy consumption, potential for high-volume hydrogen production, and smaller carbon footprint compared to thermochemical processes. A Lewis acid (FeCl₃) was used as a charge carrier and redox medium to aid in biomass depolymerisation/oxidation. A comprehensive analysis was conducted, measuring hydrogen and CO₂ emission volumes and performing electrochemical analysis (linear sweep voltammetry and chronoamperometry) to understand the process. The highest hydrogen volume was 12.1 mL, produced by FeCl₃-mediated electrolysis of glucose at ambient temperature, which was up to two times higher than starch, lignin, and cellulose at 1.20 V. Glucose also showed a maximum power-to-hydrogen yield ratio of 30.99 kWh/kg. The results showed that hydrogen can be produced from biomass feedstock at ambient temperature using FeCl₃, with higher yield and lower electricity consumption compared to water electrolysis. The study also found that the hydrogen content of the feedstock is a key factor in hydrogen production. The maximum hydrogen yields varied from 6.3 to 12.1 mL, with glucose showing the highest yield. The power-to-hydrogen yield ratio at ambient conditions ranged from 30.99 to 33.83 kWh/kg, which is more energy-efficient than conventional water electrolysis (55–58 kWh/kg). CO₂ emissions were also investigated, showing low emissions due to a maximum carbon consumption rate of around 13%. The study concludes that biomass electrolysis using FeCl₃ is a promising method for hydrogen production, with potential for cost-effective and sustainable hydrogen generation. The results suggest that biomass electrolysis can be operated at ambient temperature, with a higher yield and lower electricity consumption compared to water electrolysis. The study also highlights the importance of feedstock selection based on hydrogen content and biomass structure. The findings contribute to the development of low-carbon-emission hydrogen production from biomass resources.