Separation of sulphuric acid and glucose using steady-state recycling chromatography (SSR) was investigated experimentally and by simulations. The application is relevant for the recovery of fermentable monosaccharides from lignocellulosic hydrolysates. Choosing appropriate cut times beforehand was found to be challenging because of complex phase equilibrium behavior. An empirical isotherm model that accounts for co-operative sorption was used in simulations. SSR provided 60% higher productivity with 45% lower eluent consumption than batch when equal flow rates were used. Productivity of SSR was strongly improved without a significant increase in eluent consumption by simultaneously increasing flow rate and injection volume.