1. Expanding the Use of Dynamic Electrostatic Repulsion Reversed-Phase Chromatography: An Effective Elution Mode for Peptides Control and Analysis
- Author
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Michele Bassan, Marco Macis, Giulia Mazzoccanti, Simone Manetto, Antonio Ricci, Antonia Iazzetti, Walter Cabri, Francesco Gasparrini, and G. Mazzoccanti, S. Manetto, M. Bassan, M. Macis, A. Iazzetti, W. Cabri, A.Ricci, F. Gasparrini
- Subjects
Settore CHIM/01 - CHIMICA ANALITICA ,basic analytes ,Resolution (mass spectrometry) ,Static Electricity ,Pharmaceutical Science ,Organic chemistry ,Peptide ,01 natural sciences ,Article ,Analytical Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,QD241-441 ,Phase (matter) ,Drug Discovery ,Trifluoroacetic acid ,Physical and Theoretical Chemistry ,high peak capacity ,basic analyte ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,Chromatography, Reverse-Phase ,Chromatography ,Elution ,dynamic electrostatic repulsive reversed-phase ,peptide pharmaceuticals ,peptide pharmaceutical ,010401 analytical chemistry ,Reversed-phase chromatography ,Electrostatics ,0104 chemical sciences ,Isoelectric point ,chemistry ,Chemistry (miscellaneous) ,Molecular Medicine ,Peptides - Abstract
Bioactive peptides are increasingly used in clinical practice. Reversed-phase chromatography using formic or trifluoroacetic acid in the mobile phase is the most widely used technique for their analytical control. However, sometimes it does not prove sufficient to solve challenging chromatographic problems. In the search for alternative elution modes, the dynamic electrostatic repulsion reversed-phase was evaluated to separate eight probe peptides characterised by different molecular weights and isoelectric points. This technique, which involves TBAHSO4 in the mobile phase, provided the lowest asymmetry and peak width at half height values and the highest in peak capacity (about 200 for a gradient of 30 min) and resolution concerning the classic reversed-phase. All analyses were performed using cutting-edge columns developed for peptide separation, and the comparison of the chromatograms obtained shows how the dynamic electrostatic repulsion reversed-phase is an attractive alternative to the classic reversed-phase.
- Published
- 2021