On-line cut-off technique and organic modifier addition aided signal enhancement for trace analysis of carbohydrates in cellulase hydrolysate by ion exclusion chromatography-electrospray ionization mass spectrometry.
Paper cellulose has been hydrolyzed with calcium alginate immobilized cellulase to produce carbohydrate products and the three trace sugars, galactose, arabinose, and mannose in the cellulase hydrolysate have been analyzed by HPIEC/ESI-MS. Applying the on-line cut-off technique to the HPIEC/ESI-MS can cut the high concentration glucose off to eliminate its interference on the peaks of minor sugars and enhance their signals from 1.1- to 1.6-fold. However, the on-line post column addition of 15% ethanol to the eluate can increase the signal of the three trace sugars, galactose, arabinose, and mannose up to 17-, 23-, and 11-fold, respectively, and make the corresponding detection limits as 0.04, 0.04, and 0.03 ppm. The accuracies of the quantitative analysis for the three trace sugars with the signal enhanced HPIEC/ESI-MS by the two enhancement methods were larger than 95%. The precisions of the analytical results were also greatly improved by the assistance of the two techniques and were less than 6.5%. The quantitative analysis of the three trace sugars was performed with the internal standard method and the internal standard (IS) was sorbitol. Overall, the signal enhancement of HPIEC/ESI-MS and quantification of the three trace sugars by the on-line cut-off technique and organic modifier addition was successful.
Cheng
Cheanyeh
C
Department of Chemistry, Chung Yuan Christian University, Chungli, Taiwan, Republic of China. chengce@cycu.edu.tw
Tsai
Hsiang-Rong
HR
Chang
Kuo-Chung
KC
D001089:Arabinose / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis*; D002480:Cellulase / Q000378:metabolism*; D002482:Cellulose / Q000737:chemistry / Q000378:metabolism; D002850:Chromatography, Gel / Q000379:methods*; D004800:Enzymes, Immobilized; D005690:Galactose / Q000032:analysis; D008358:Mannose / Q000032:analysis; D009862:Online Systems; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D002241:Carbohydrates; D004800:Enzymes, Immobilized; D002482:Cellulose; D001089:Arabinose; D002480:Cellulase; D008358:Mannose; D005690:Galactose