Effect of Cyclodextrin Complexation on the Separation of Insulin Analogues in Capillary Electrophoresis Using Dynamically Coated Capillaries
6 affiliations
- 1Department of Physical Chemistry, “George Emil Palade” University of Medicine, Pharmacy, Science, and Technology of Targu Mures, 540142 Târgu Mureş, Romania
- 2Doctoral School of Medicine and Pharmacy, “George Emil Palade” University of Medicine, Pharmacy, Science, and Technology of Targu Mures, 540142 Târgu Mureş, Romania
- 3Department of Pharmaceutical Industry and Biotechnology, “George Emil Palade” University of Medicine, Pharmacy, Science, and Technology of Targu Mures, 540142 Târgu Mureş, Romania
- 4Department of Pharmaceutical Chemistry, Semmelweis University, Hőgyes E. u. 9, 1092 Budapest, Hungary
- 5Center for Pharmacology and Drug Research & Development, Semmelweis University, 1092 Budapest, Hungary
- 6Sz-imfidum Ltd., 525401 Lunga, Romania
Abstract
The simultaneous separation of therapeutic insulin analogues by capillary electrophoresis (CE) remains challenging because of their high structural similarity and their tendency to adsorb onto the capillary surface. This study investigated the applicability of cyclodextrin (CD) derivatives as selectivity-modifying additives for the CE separation of human insulin and five therapeutic insulin analogues using five-layer poly(diallyldimethylammonium chloride)/poly(sodium 4-styrenesulfonate) (PDADMAC/PSS)2.5 and four-layer (PDADMAC/PSS)2 successive multiple ionic-polymer layer (SMIL)-coated capillaries under acidic and alkaline conditions, respectively. A broad range of CD derivatives was systematically evaluated under optimized background electrolyte conditions. Under acidic conditions, neutral and predominantly uncharged CD derivatives improved separation selectivity, with carboxyethyl-β-CD providing the most favorable discrimination within the critical insulin lispro–human insulin–insulin aspart migration group. In contrast, highly charged anionic CD derivatives disrupted the PDADMAC-terminated coating and could not be applied. The PSS-terminated coating remained stable under alkaline conditions, enabling the evaluation of both neutral and anionic CD derivatives. Under these conditions, the migration order of the insulin species was completely reversed, while several CD derivatives further modulated the relative spacing of neighboring peaks. The use of organic modifiers produced system-dependent effects on separation selectivity and peak shape. These findings demonstrate that the appropriate selection of CD derivatives and SMIL coating strategy provides an effective approach for modulating selectivity in the CE separation of structurally related therapeutic peptides.
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BibTeX
@article{kis2026effect,
title = {{Effect of Cyclodextrin Complexation on the Separation of Insulin Analogues in Capillary Electrophoresis Using Dynamically Coated Capillaries}},
author = {Csenge Kis and Emese Sipos and Gergő Tóth and Zoltán-István Szabó},
journal = {Molecules (Basel, Switzerland)},
year = {2026},
month = oct,
doi = {10.3390/molecules31193578},
url = {https://europepmc.org/article/PMC/PMC13667192},
note = {CC BY},
}
RIS
TY - JOUR
TI - Effect of Cyclodextrin Complexation on the Separation of Insulin Analogues in Capillary Electrophoresis Using Dynamically Coated Capillaries
AU - Csenge Kis
AU - Emese Sipos
AU - Gergő Tóth
AU - Zoltán-István Szabó
PY - 2026
DA - 2026/10/01
JO - Molecules (Basel, Switzerland)
DO - 10.3390/molecules31193578
UR - https://europepmc.org/article/PMC/PMC13667192
AB - The simultaneous separation of therapeutic insulin analogues by capillary electrophoresis (CE) remains challenging because of their high structural similarity and their tendency to adsorb onto the capillary surface. This study investigated the applicability of cyclodextrin (CD) derivatives as selectivity-modifying additives for the CE separation of human insulin and five therapeutic insulin analogues using five-layer poly(diallyldimethylammonium chloride)/poly(sodium 4-styrenesulfonate) (PDADMAC/PSS)2.5 and four-layer (PDADMAC/PSS)2 successive multiple ionic-polymer layer (SMIL)-coated capillaries under acidic and alkaline conditions, respectively. A broad range of CD derivatives was systematically evaluated under optimized background electrolyte conditions. Under acidic conditions, neutral and predominantly uncharged CD derivatives improved separation selectivity, with carboxyethyl-β-CD providing the most favorable discrimination within the critical insulin lispro–human insulin–insulin aspart migration group. In contrast, highly charged anionic CD derivatives disrupted the PDADMAC-terminated coating and could not be applied. The PSS-terminated coating remained stable under alkaline conditions, enabling the evaluation of both neutral and anionic CD derivatives. Under these conditions, the migration order of the insulin species was completely reversed, while several CD derivatives further modulated the relative spacing of neighboring peaks. The use of organic modifiers produced system-dependent effects on separation selectivity and peak shape. These findings demonstrate that the appropriate selection of CD derivatives and SMIL coating strategy provides an effective approach for modulating selectivity in the CE separation of structurally related therapeutic peptides.
SN - 1420-3049
ER -