On-line desalting and carbohydrate analysis for immobilized enzyme hydrolysis of waste cellulosic biomass by column-switching high-performance liquid chromatography.
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 0-2,000 microg 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 microg 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.
Cheng
Cheanyeh
C
Department of Chemistry, Chung Yuan Christian University, 200 Chung Pei Road, Chungli 32023, ROC Taiwan. chengce@cycu.edu.tw
Chen
Chi-Sung
CS
Hsieh
Pei-Hsin
PH
D018533:Biomass; D002138:Calibration; D002241:Carbohydrates / Q000032:analysis*; D002482:Cellulose / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D016018:Least-Squares Analysis; D010209:Paper; D035261:Plant Components, Aerial / Q000378:metabolism; D012015:Reference Standards; D012037:Refuse Disposal; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D002241:Carbohydrates; D004800:Enzymes, Immobilized; D002482:Cellulose