A Density‐Corrected Method for Accurate Glucose Quantification During High‐Solids Enzymatic Hydrolysis of Corn

ABSTRACT Standard glucose assays in high‐solids enzymatic hydrolysis often ignore hydrolysate density, which inflates concentrations and yields. To address this critical flaw, this research develops and validates a density‐corrected calculation that incorporates the measured hydrolysate density and quantify the error associated with the common, density‐ignored approach. Theoretical modeling predicted a substantial overestimation by the uncorrected method, a finding confirmed during the 40% solids hydrolysis of extruded corn. Experimentally, the uncorrected method progressively overestimated the true glucose concentration, with the discrepancy reaching approximately 15% after 24 h. This resulted in a significantly inflated apparent glucose yield of 106.5% compared to the plausible 92.9% yield calculated with the corrected calculation. A theoretical analysis across 5%–50% solids loading shows the discrepancy grows with loading and remains robust to assumptions about minor soluble non‐starch components. The results demonstrate that density effects are a primary source of analytical error under the high‐titer conditions targeted for process intensification. Incorporating the measured hydrolysate density restores mass‐balanced concentrations and yields, enabling reliable monitoring and fair benchmarking of high‐solids starch conversion for food ingredients and bioprocess feedstocks.