On-line desalting and carbohydrate analysis for immobilized enzyme hydrolysis of waste cellulosic biomass by column-switching high-performance liquid chromatography

Abstract

An innovative green column-switching high-performance liquid chromatographic (HPLC) technique was developed by coupling traditional and Pb 2+ ion-exclusion columns to study enzyme hydrolysis components of waste cellulosic biomass. Pure water was used as the mobile phase to separate neutral polar analytes in high salt content solution. The column-switching HPLC-RI was connected on-line to the immobilized enzyme reactor for successive on-line desalting and simultaneous analysis of six carbohydrates (cellobiose, glucose, xylose, galactose, mannose, and arabinose) in the hydrolysate of waste paper and waste tree branch by incorporating the heart-cut and the elution-time-difference techniques. Six internal standard calibration curves in the linear concentration range of 0–2000 μg mL −1 were prepared. Xylitol was used as the internal standard to give excellent linear correlation coefficients (0.9984–0.9999). The limits of detection and quantification for cellobiose, glucose, xylose, galactose, mannose, and arabinose varied between 0.12–4.88 and 0.40–16.3 μg mL −1, respectively, with an accuracy of 90–102% and a precision of 0.1–7.8%. Cellulose and hemicellulose contents were higher in waste paper than in waste tree branch.