Cationic Phenylpyridine Labeling for Improved Oligosaccharide and Glycan Analysis by Capillary…

Abstract

Capillary electrophoresis with laser-induced fluorescence detection (CE/LIF) is a powerful technique for analyzing glycans. However, it requires derivatization with a fluorescent compound to enhance both electrophoretic mobility and detection sensitivity simultaneously. The widely used labeling reagent 8-aminopyrene-1,3,6-trisulfonic acid (APTS) suffers from several limitations, including relatively low labeling efficiency and poor compatibility with mass spectrometry (MS), highlighting the need for improved alternatives. In this work, we designed and synthesized a novel cationic fluorescent label based on a phenylpyridine scaffold incorporating hydrazide functionality for efficient oligosaccharide and glycan derivatization via hydrazone formation. To fully exploit its fluorescence characteristics, we also optimized the capillary electrophoresis system by modifying the LED-induced fluorescence (LED-IF) detector through systematic evaluation of excitation LEDs and emission filters, resulting in substantially improved detection sensitivity. The optimized labeling, separation, and detection conditions enabled sensitive analysis of released glycans from various glycoproteins with detection limits in the nanomolar concentration range. The combination of this new phenylpyridine-based fluorescent label with the enhanced CE/LED-IF detection platform significantly improves the analytical performance of CE for glycan analysis and provides a more sensitive, robust, and versatile approach for comprehensive oligosaccharide and glycan characterization.

Choosing the best capillary coatings based on mixtures of cationic and neutral double-chained surfactants for protein separation by capillary electrophoresis
Chirapha Prakobdi, Laurent Leclercq, Phoonthawee Saetear +1
Analytica Chimica Acta24 Oct 2026Peer-reviewedCC BY 4.0Full textAnalytical chemistryHAL
A Mixed-Micelle Capillary Electrophoresis Method for Fast Quality Screening of Antimalarial Drugs
Hailekiros Gebretsadik Kidanemariam, Erwin Adams, Ann Van Schepdael
Molecules28 Oct 2026Peer-reviewedCC BY 4.0Analytical chemistryCrossref
Vancomycin-Stabilized Copper Nanoclusters: Isolation by Solid-Phase Extraction and Application in Chiral Capillary Electrophoresis
Ekaterina A. Kolobova, Andrei I. Demenshin, Yulia P. Zimarina +1
Nanomaterials26 Oct 2026Peer-reviewedCC BY 4.0Physical chemistryCrossref
Clindamycin Phosphate-Mediated Enantioseparation of Amino Acids by Capillary Electrophoresis: Molecular Modeling and NMR Insights into the Chiral Recognition Mechanism
Mohammed A. A. Khalid, Abdalla A. Elbashir, Munirah A. Alfarhan +1
Molecules1 Oct 2026Peer-reviewedCC BY 4.0Full textPhysical chemistryCrossrefEurope PMC
Effect of Cyclodextrin Complexation on the Separation of Insulin Analogues in Capillary Electrophoresis Using Dynamically Coated Capillaries
Csenge Kis, Emese Sipos, Gergő Tóth +1
Molecules (Basel, Switzerland)1 Oct 2026Peer-reviewedCC BYFull textGeneral chemistryEurope PMCCrossref

Cite

BibTeX RIS

BibTeX

@article{dusa2026cationic,
  title = {{Cationic Phenylpyridine Labeling for Improved Oligosaccharide and Glycan Analysis by Capillary Electrophoresis with LED-Induced Fluorescence Detection}},
  author = {Filip Dusa and Michal Gregus and Richard Cmelik and Hubert Vesely and Hana Pizova and Pavel Bobal and Jana Lavicka},
  journal = {ACS Omega},
  year = {2026},
  month = nov,
  doi = {10.1021/acsomega.6c08725},
  url = {https://pubs.acs.org/acsodf/article/doi/10.1021/acsomega.6c08725/5466275/Cationic-Phenylpyridine-Labeling-for-Improved},
  note = {CC BY 4.0},
}

RIS

TY  - JOUR
TI  - Cationic Phenylpyridine Labeling for Improved Oligosaccharide and Glycan Analysis by Capillary Electrophoresis with LED-Induced Fluorescence Detection
AU  - Filip Dusa
AU  - Michal Gregus
AU  - Richard Cmelik
AU  - Hubert Vesely
AU  - Hana Pizova
AU  - Pavel Bobal
AU  - Jana Lavicka
PY  - 2026
DA  - 2026/11/08
JO  - ACS Omega
DO  - 10.1021/acsomega.6c08725
UR  - https://pubs.acs.org/acsodf/article/doi/10.1021/acsomega.6c08725/5466275/Cationic-Phenylpyridine-Labeling-for-Improved
AB  - Capillary electrophoresis with laser-induced fluorescence detection (CE/LIF) is a powerful technique for analyzing glycans. However, it requires derivatization with a fluorescent compound to enhance both electrophoretic mobility and detection sensitivity simultaneously. The widely used labeling reagent 8-aminopyrene-1,3,6-trisulfonic acid (APTS) suffers from several limitations, including relatively low labeling efficiency and poor compatibility with mass spectrometry (MS), highlighting the need for improved alternatives. In this work, we designed and synthesized a novel cationic fluorescent label based on a phenylpyridine scaffold incorporating hydrazide functionality for efficient oligosaccharide and glycan derivatization via hydrazone formation. To fully exploit its fluorescence characteristics, we also optimized the capillary electrophoresis system by modifying the LED-induced fluorescence (LED-IF) detector through systematic evaluation of excitation LEDs and emission filters, resulting in substantially improved detection sensitivity. The optimized labeling, separation, and detection conditions enabled sensitive analysis of released glycans from various glycoproteins with detection limits in the nanomolar concentration range. The combination of this new phenylpyridine-based fluorescent label with the enhanced CE/LED-IF detection platform significantly improves the analytical performance of CE for glycan analysis and provides a more sensitive, robust, and versatile approach for comprehensive oligosaccharide and glycan characterization.
SN  - 2470-1343
ER  -

Abstract

Related

Cite