Quantification of Sugar Compounds and Uronic Acids in Enzymatic Hydrolysates of Lignocellulose Using High-Performance Anion Exchange Chromatography with Pulsed Amperometric Detection

Abstract

A method was developed for simultaneous determination of seven sugar compounds and two uronic acids by high-performance anion exchange chromatography (HPAEC) with pulsed amperometric detection (PAD) using a CarboPac PA20 column under a gradient elution (mobile phase: H 2 O/200 mM NaOH/100 mM NaAc) at 0.4 mL/min. Under these conditions, D-(−)-arabinose, D-(+)-galactose, D-(+)-glucose, D-(+)-mannose, D-(+)-xylose, D-(−)-fructose, D-(+)-cellobiose, Dgalacturonic acid, and D-glucuronic acid were all separated in 46 min. The calibration curve, detection limits, reproducibility and accuracy were also determined. Furthermore, seven sugar compounds and two uronic acids were all detected in the enzymatic hydrolysates of different lignocellulosic materials treated by different pretreatment processes. Our results demonstrate that this method was feasible for sugar compounds separation without the need for derivatization and organic solvent, and it also has the advantages of having low interference, high sensitivity, low detection limit of picomol, and excellent reproducibility and accuracy.

New Method for Carbohydrates Determination in Sugarcane Bagasse by HPAEC‐RPAD Using Glassy Carbon Electrode Modified with Carbon Nanotubes and Nickel Nanoparticles

“…The results are comparable to those of other studies reported in the literature with regard to the analytical determination of carbohydrates by chromatographic methods for carbohydrates obtained from biomass samples. Compared with several current electroanalytical techniques for the assay of carbohydrates, the HPAEC‐RPAD technique bears a great advantage as a simultaneous analysis technique.…”

“…The underlying shortcoming of these methods is that they require sample derivatization and this somewhat renders the analysis quite unwieldy . As a result, HPAEC‐High Performance Anion‐Exchange Chromatography has been more frequently employed compared to GC when it comes to carbohydrate determination in biomass.…”

Determination of uronic acids in sugarcane bagasse by anion-exchange chromatography using an electrode modified with copper nanoparticles

“…11,12,22,23 In general, the sequence of elution of the matrix components is related to the pK a value. 11,12,22,23 In general, the sequence of elution of the matrix components is related to the pK a value.…”

“…6 Traditional methods for the detection of uronic acids are based on colorimetry using carbazole-sulfuric acid and meta-hidroxydiphenyl, 7,8 derivatizing the uronic acids to be detected. 10,11 However, this method involves a long run time, multiple dilutions of samples, and use of multiple pumps to run in gradient (consequently using large amounts of solvents for separation) and it requires the addition of a strong hydroxide post-column to minimize baseline dri. 9 Several methods have been used for the determination of sugars as well as uronic acids in lignocellulosic waste, including gas chromatography, high-performance liquid chromatography, and capillary electrophoresis.…”

“…The studies using working electrodes modied with copper have shown that several electroactive organic molecules can be determined with good sensitivity in order to improve the performance and reduce the undesirable amperometric poisoning. 9,22 Furthermore, the method has the advantage of being simpler, with high sensitivity and low detection limit. [11][12][13] Separation is bound to occur based on the difference between the ion exchange uronic acids and the stationary phase.…”