Characterization and comparison of mixed-mode and reversed-phase columns; interaction abilities and applicability for peptide separation

Kadlecová, Zuzana; Kozlík, Petr; Tesařová, Eva; Gilár, Martin; Kalíková, Květa

doi:10.1016/j.chroma.2021.462182

Cited by 17 publications

(11 citation statements)

References 53 publications

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“…The structure of the 1.7 μm BEH particles also uses novel final protection processes, improving the shape of the peak for basic analytes, which increases the sensitivity of LC-MS analysis and simplifies sample preparation [34]. On the other hand, the 5 μm Luna Omega PS C18 column contains different ligands that give it a broad polar and non-polar retention [35]. However, the exact composition of the stationary phase of this column is not available as it is patented information.…”

Section: Optimization Of the Hplc-apci-qqq-ms/ms Methodsmentioning

confidence: 99%

Development of a QuEChERS method for simultaneous analysis of 3-Monochloropropane-1,2-diol monoesters and Glycidyl esters in edible oils and margarine by LC-APCI-MS/MS

Custodio-Mendoza

Sendón

Quirós

et al. 2023

Analytica Chimica Acta

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Section: Optimization Of the Hplc-apci-qqq-ms/ms Methodsmentioning

confidence: 99%

Development of a QuEChERS method for simultaneous analysis of 3-Monochloropropane-1,2-diol monoesters and Glycidyl esters in edible oils and margarine by LC-APCI-MS/MS

Custodio-Mendoza

Sendón

Quirós

et al. 2023

Analytica Chimica Acta

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“…This particle chemistry was introduced by Waters in 2010 under a trademark CSH (Charged Surface Hybrid) . CSH particles are covered by a low amount of ionizable pyridyl groups before they are functionalized by ligands for RPLC . At a pH < 6, the pyridyl ligands are positively charged .…”

Section: Rplc Methods For Bottom-up Proteomic Analysesmentioning

confidence: 99%

“…281 CSH particles are covered by a low amount of ionizable pyridyl groups before they are functionalized by ligands for RPLC. 282 At a pH < 6, the pyridyl ligands are positively charged. 283 Therefore, positively charged peptides are repelled from the proximity of residual silanols by electrostatic repulsion under acidic pH, improving their peak shape and loadability in the low-ionicstrength mobile phase.…”

Section: Journal Of Proteome Researchmentioning

confidence: 99%

Reversed-Phase Liquid Chromatography of Peptides for Bottom-Up Proteomics: A Tutorial

et al. 2022

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The performance of the current bottom-up liquid chromatography hyphenated with mass spectrometry (LC-MS) analyses has undoubtedly been fueled by spectacular progress in mass spectrometry. It is thus not surprising that the MS instrument attracts the most attention during LC-MS method development, whereas optimizing conditions for peptide separation using reversed-phase liquid chromatography (RPLC) remains somewhat in its shadow. Consequently, the wisdom of the fundaments of chromatography is slowly vanishing from some laboratories. However, the full potential of advanced MS instruments cannot be achieved without highly efficient RPLC. This is impossible to attain without understanding fundamental processes in the chromatographic system and the properties of peptides important for their chromatographic behavior. We wrote this tutorial intending to give practitioners an overview of critical aspects of peptide separation using RPLC to facilitate setting the LC parameters so that they can leverage the full capabilities of their MS instruments. After briefly introducing the gradient separation of peptides, we discuss their properties that affect the quality of LC-MS chromatograms the most. Next, we address the in-column and extra-column broadening. The last section is devoted to key parameters of LC-MS methods. We also extracted trends in practice from recent bottom-up proteomics studies and correlated them with the current knowledge on peptide RPLC separation.

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“…Kadlecová et al [ 103 ] compared two columns with positively charged ionic ligands (XSelect CSH C18, containing pyridyl group, and Atlantis PREMIER BEH C18 AX, containing a quaternary alkylamine) for the separation of 14 peptides, especially ten dipeptides, three pentapeptides and one octapeptide. The analytes have been separated in gradient conditions using ammonium formate 5 mM (pH 3) and ACN.…”

Section: Purification Techniques (Downstream Processing)mentioning

confidence: 99%

Downstream Processing of Therapeutic Peptides by Means of Preparative Liquid Chromatography

et al. 2021

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The market of biomolecules with therapeutic scopes, including peptides, is continuously expanding. The interest towards this class of pharmaceuticals is stimulated by the broad range of bioactivities that peptides can trigger in the human body. The main production methods to obtain peptides are enzymatic hydrolysis, microbial fermentation, recombinant approach and, especially, chemical synthesis. None of these methods, however, produce exclusively the target product. Other species represent impurities that, for safety and pharmaceutical quality reasons, must be removed. The remarkable production volumes of peptide mixtures have generated a strong interest towards the purification procedures, particularly due to their relevant impact on the manufacturing costs. The purification method of choice is mainly preparative liquid chromatography, because of its flexibility, which allows one to choose case-by-case the experimental conditions that most suitably fit that particular purification problem. Different modes of chromatography that can cover almost every separation case are reviewed in this article. Additionally, an outlook to a very recent continuous chromatographic process (namely Multicolumn Countercurrent Solvent Gradient Purification, MCSGP) and future perspectives regarding purification strategies will be considered at the end of this review.

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Characterization and comparison of mixed-mode and reversed-phase columns; interaction abilities and applicability for peptide separation

Cited by 17 publications

References 53 publications

Development of a QuEChERS method for simultaneous analysis of 3-Monochloropropane-1,2-diol monoesters and Glycidyl esters in edible oils and margarine by LC-APCI-MS/MS

Development of a QuEChERS method for simultaneous analysis of 3-Monochloropropane-1,2-diol monoesters and Glycidyl esters in edible oils and margarine by LC-APCI-MS/MS

Reversed-Phase Liquid Chromatography of Peptides for Bottom-Up Proteomics: A Tutorial

Downstream Processing of Therapeutic Peptides by Means of Preparative Liquid Chromatography

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