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Capillary Zone Electrophoresis of 8-Aminopyrene-1,3,6-trisulfonic Acid Labeled Carbohydrates with Online Electrokinetic Sample Cleanup. | LitMetric

Capillary Zone Electrophoresis of 8-Aminopyrene-1,3,6-trisulfonic Acid Labeled Carbohydrates with Online Electrokinetic Sample Cleanup.

Anal Chem

Translational Glycomics Group, Research Institute of Biomolecular and Chemical Engineering, University of Pannonia, H-8200 Veszprem, Hungary.

Published: November 2023

AI Article Synopsis

  • Capillary electrophoresis is a technique used to separate complex carbohydrates, but sugars need to be tagged with a charged fluorophore for analysis, which often requires additional purification steps that can be labor-intensive and costly.
  • A new online electrokinetic sample cleanup process using electroosmotic flow was proposed, enabling separation without the need for extra purification step, thus reducing the risk of sample loss.
  • The study demonstrated that using a specific running buffer (caproic acid-Tris at pH 8.1) effectively separates components of interest from excess labeling reagents when analyzing human serum samples, resulting in excellent separation performance.

Article Abstract

Capillary electrophoresis is one of the frequently used separation techniques for the analysis of complex carbohydrates. Since sugars lack chromophore or fluorophore groups, their capillary electrophoresis analysis usually requires tagging by a charged fluorophore. To speed up the derivatization reaction, a large excess of the labeling reagent is typically used; therefore, a purification step is necessary prior to CE analysis using the industry standard low-pH gel-buffer system. In addition to representing an extra sample preparation step with the associated labor and cost, the purification process also holds the risk of losing some of the sample components. In this paper we introduce an online electrokinetic sample cleanup process with electroosmotic flow (EOF)-assisted separation in a bare fused silica capillary using alkaline pH background electrolyte and normal polarity separation voltage. 8-Aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled maltooligosaccharides were analyzed first to understand the complex effect of the downstream EOF and the counter current electromigration of the sample components including the labeling dye. The use of 150 mM caproic acid-253 mM Tris (pH 8.1) running buffer facilitated the entrance of the sample components of interest into the separation capillary, while the excess labeling reagent was excluded and, therefore, did not interfere with the detection. The alkaline caproic acid-Tris running buffer was then applied to the N-glycome analysis of human serum samples, showing excellent separation performance, and more importantly, the extra sample purification step was not required.

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Source
http://dx.doi.org/10.1021/acs.analchem.3c03714DOI Listing

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