Sampling, Dilution, and Loading Device-coupled HighPerformance Liquid Chromatography Method for Successive On-line Analyses of Major Carbohydrate Products in Immobilized Cellulase Hydrolysate of Paper Cellulose

Abstract

A device with direct sampling, dilution, and sample loading has been utilized to couple a vertically hanging immobilized cellulase bioreactor with a high-performance liquid chromatography system to make a successful on-line analysis of the 3 major carbohydrate products: glucose, xylose, and cellobiose in the cellulase hydrolysate. The coupled analysis system for successive on-line monitoring of the enzymatic hydrolysis of paper cellulose was stable over a period of 96 h. A comparative study involving 4 different quantification methods for the 3 carbohydrates was conducted. The internal standard calibration method with on-line dilution was observed to be the best procedure in terms of accuracy, precision, limit of detection, and systematic error reduction. With this method, the analysis for glucose had 99.9% accuracy, RSD below 2.4%, and a 2.1 ppm detection limit. Matrix matching was found important in the preparation of standard calibration curves with refractive index detection.