Sampling, dilution, and loading device-coupled high-performance liquid chromatography method for successive on-line analyses of major carbohydrate products in immobilized cellulase hydrolysate of paper cellulose.
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.
Cheng
Cheanyeh
C
Department of Chemistry, Chung Yuan Christian University, Chungli, Taiwan, Republic of China. chengce@cycu.edu.tw
Chang
Kuo-Chung
KC
D002241:Carbohydrates / Q000032:analysis* / Q000737:chemistry; D002480:Cellulase / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D009862:Online Systems
D002241:Carbohydrates; D004800:Enzymes, Immobilized; D002482:Cellulose; D002480:Cellulase