The imminent use of hydrogen as an energy vector establishes the need for sustainable production technologies based on renewable resources. Starch is an abundant renewable resource suitable for bio-hydrogen generation. It was hypothesised that starch hydrolysates from a large (250 mL) hydrothermal reactor could support bioH 2 fermentation without inhibition by toxic byproducts.Starch was hydrolysed at high concentrations (40-200 g.L -1 ) in hot compressed water (HCW) with CO 2 at 30 bar in a 250 mL reactor, the largest so far for polysaccharide hydrolysis, at 180-235 °C, 15 min. Hydrolysates were detoxified with activated carbon (AC) and tested in biohydrogen fermentations. The maximum yield of glucose was 548 g.kg starch -1 carbon at 200 °C. 5-hydroxymethyl furfural, the main fermentation inhibitor, was removed by AC to support 70% more hydrogen production than the untreated hydrolysates. The potential utilization of starch hydrolysates from HCW treatment for upscaled fermentations is promising.