Evaluation of solid-phase extraction sorbents for purification of oligosaccharides and glycans derivatized by positively charged labels followed by capillary electrophoretic analysis

0580317 - ÚIACH 2025 RIV US eng J - Journal Article
Smolková, Denisa - Moravcová, Dana - Lavická, Jana
Evaluation of solid-phase extraction sorbents for purification of oligosaccharides and glycans derivatized by positively charged labels followed by capillary electrophoretic analysis.
Journal of Separation Science. (2024), s. 1-11. ISSN 1615-9306. E-ISSN 1615-9314
R&D Projects: GA ČR(CZ) GA22-00236S
Institutional support: RVO:68081715
Keywords : sample preparation * protein glycosylation * n-glycans * fluorescence * capillary electrophoresis * solid-phase extraction
OECD category: Analytical chemistry
Impact factor: 3.1, year: 2022
Method of publishing: Open access
https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/jssc.202300705

The sample preparation including labeling and clean-up represents a key analytical step in the analysis of oligosaccharides and glycans by either chromatographic or electrophoretic separation methods. Although the majority of labeling has been performed by neutral and/or negatively charged tags, the introduction of a positive charge into the saccharide molecule can significantly improve the analysis, especially with mass spectrometry detection. In this work, we present the evaluation of five solid-phase extraction sorbents differing in extraction chemistry for the clean-up and concentration of positively labeled maltooligosaccharides from the reaction mixtures. Maltooligosaccharides containing four to seven glucose units were labeled by cationic tags (2-aminoethyl)trimethylammonium chloride and (carboxymethyl)trimethylammonium chloride hydrazide and the extraction conditions were optimized followed by electrophoretic analysis with conductivity detection. The effects of the solid-phase extraction sorbent chemistry, extraction conditions, and sample composition are discussed. All tested sorbents were capable of cleaning up maltooligosaccharides from the reaction mixtures to some extent after optimization of the solid-phase extraction procedure (51.9%-98.9% recovery). The best-rated amide-based sorbent was used to process the sample of N-linked glycans enzymatically released from ribonuclease B.
Permanent Link: https://hdl.handle.net/11104/0349082