High–throughput enantioseparation of Nα–fluorenylmethoxycarbonyl proteinogenic amino acids through fast chiral chromatography on zwitterionic-teicoplanin stationary phases

Mazzoccanti, Giulia; Manetto, Simone; Ricci, Antônio; Cabri, Walter; Orlandin, Andrea; Catani, Martina; Felletti, Simona; Cavazzini, Alberto; Ye, Micheal; Ritchie, Harald; Villani, Claudio; Gasparrini, Francesco

doi:10.1016/j.chroma.2020.461235

Cited by 22 publications

(14 citation statements)

References 39 publications

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“…The particle diameters also seem to influence the enantioseparation. Indeed, the morphology differences of fully-and superficially-porous particles (FPP and SPP, respectively) affect the enantioselectivity and the resolution, in favour of the SPP column [100].…”

Section: Macrocycle Antibiotic-based Chiral Stationary Phasesmentioning

confidence: 99%

Recent Advances in Chiral Analysis of Proteins and Peptides

Morvan

Mikšı́k

2021

Separations

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Like many biological compounds, proteins are found primarily in their homochiral form. However, homochirality is not guaranteed throughout life. Determining their chiral proteinogenic sequence is a complex analytical challenge. This is because certain D-amino acids contained in proteins play a role in human health and disease. This is the case, for example, with D-Asp in elastin, β-amyloid and α-crystallin which, respectively, have an action on arteriosclerosis, Alzheimer's disease and cataracts. Sequence-dependent and sequence-independent are the two strategies for detecting the presence and position of D-amino acids in proteins. These methods rely on enzymatic digestion by a site-specific enzyme and acid hydrolysis in a deuterium or tritium environment to limit the natural racemization of amino acids. In this review, chromatographic and electrophoretic techniques, such as LC, SFC, GC and CE, will be recently developed (2018–2020) for the enantioseparation of amino acids and peptides. For future work, the discovery and development of new chiral stationary phases and derivatization reagents could increase the resolution of chiral separations.

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Section: Macrocycle Antibiotic-based Chiral Stationary Phasesmentioning

confidence: 99%

Recent Advances in Chiral Analysis of Proteins and Peptides

Morvan

Mikšı́k

2021

Separations

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“…reaction for the linkage of the chiral selector to the surface of silica sphere [84]. However, it is strange that just a few articles related to ICSs in HPLC are reported in recent 3 years [55,60,61,[84][85][86]. It is mainly due to the reason that the addition of ionic selectors cannot improve the recognition efficiency obviously compared with CE.…”

Section: Chromatographymentioning

confidence: 99%

Recent advances of the ionic chiral selectors for chiral resolution by chromatography, spectroscopy and electrochemistry

Fan

et al. 2021

J of Separation Science

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Ionic chiral selectors have been received much attention in the field of asymmetric catalysis, chiral recognition, and preparative separation. It has been shown that the addition of ionic chiral selectors can enhance the recognition efficiency dramatically due to the presence of multiple intermolecular interactions, including hydrogen bond, π-π interaction, van der Waals force, electrostatic ion-pairing interaction, and ionic-hydrogen bond. In the initial research stage of the ionic chiral selectors, most of work center on the application in chromatographic separation (capillary electrophoresis, high-performance liquid chromatography, and gas chromatography). Differently, more and more attention has been paid on the spectroscopy (nuclear magnetic resonance, fluorescence, ultraviolet and visible absorption spectrum, and circular dichroism spectrum) and electrochemistry in recent years. In this tutorial review as regards the ionic chiral selectors, we discuss in detail the structural features, properties, and their application in chromatography, spectroscopy, and electrochemistry.

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“…They are also present in (natural, synthetic, and therapeutic) peptides, requiring their analytical determination as part of structure elucidation and/or quality control . A direct enantioresolution of underivatized and derivatized AAs using chiral stationary phases is nowadays the first choice. − Common examples for achiral precolumn derivatization supporting chiral separation comprise 6-aminoquinolyl- N -hydroxysuccinimidyl carbamate (AQC), , 1-fluoro-2,4-dinitrobenzene (DNB-F, Sanger’s reagent), 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD), dansyl chloride (Dns), , and 9-fluorenylmethyl-chloroformate (Fmoc-Cl). − These reagents commonly improve the retention and enantioselective behavior, , introduce chromophoric and fluorophoric detectability, can enhance detection sensitivity of AAs with poor electrospray ionization efficiency, and deliver signature ions (in MS/MS detection). , Chiral stationary phases, the workhorses for direct HPLC enantiomer separation, have often remarkable enantioselectivity, but suffer from limited chemoselectivity and lower efficiencies (compared to highly efficient RP congeners). , Hence, the combination of the two principles is quite obvious and should lead to an enhanced separation technology platform.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive Online Reversed-Phase × Chiral Two-Dimensional Liquid Chromatography-Mass Spectrometry with Data-Independent Sequential Window Acquisition of All Theoretical Fragment-Ion Spectra-Acquisition for Untargeted Enantioselective Amino Acid Analysis

2022

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This work presents an advanced analytical platform for untargeted enantioselective amino acid analysis (eAAA) by comprehensive achiral × chiral 2D-LC hyphenated to ESI-QTOF-MS/MS utilizing data-independent SWATH (sequential window acquisition of all theoretical fragment-ion spectra) technology. The methodology involves N-terminal pre-column derivatization with 6-aminoquinolyl-Nhydroxysuccinimidyl carbamate (AQC; AccQ) as retention, selectivity, and MS tag, supporting retention and UV detection in RPLC ( 1 D), chiral recognition, and thus enantioselectivity by the core−shell tandem column composed of a quinine carbamate weak anion exchanger (QN-AX) and a zwitterionic chiral ion-exchanger (ZWIX(+)) ( 2 D) as well as the ionization efficiency during positive electrospray ionization due to a high proton affinity of the AQC label. Furthermore, the urea-type MS tag gives rise to the generation of AQC-tag characteristic signature fragments in MS 2 . The latter allows the chemoselective mass spectrometric filtering of targeted and untargeted Nderivatized amino acids or related labeled species. The chiral core−shell tandem column provides a complete enantioselective amino acid profile of all proteinogenic amino acids within 1 min, with full baseline separation of all enantiomers, but without resolution of isomeric Ile/allo-Ile (aIle)/Leu, which can be resolved by RPLC. The entire LC × LC separation occurs within a total run time of 60 min ( 1 D), with the chiral 2 D operated in gradient elution mode and a cycle time of 60 s. A strategy to mine the 2D-LC-SWATH data is presented and demonstrated for the qualitative eAAA of two peptide hydrolysate samples of therapeutic peptides containing common and uncommon as well as primary and secondary amino acids. Absolute configuration assignment of amino acids using template matching for all proteinogenic amino acids was made feasible due to method robustness and the inclusion of an isotopically labeled L-[U-13 C 15 N]-AA standard. The quantification performance of this LC × LC−MS/MS assay was also evaluated. Accuracies were acceptable for the majority of AAs enabling AA composition determination in peptide hydrolysates simultaneously with configuration assignment, as exemplified by oxytocin. This methodology represents a step toward truly untargeted 2D enantioselective amino acid analysis and metabolomics.

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High–throughput enantioseparation of Nα–fluorenylmethoxycarbonyl proteinogenic amino acids through fast chiral chromatography on zwitterionic-teicoplanin stationary phases

Cited by 22 publications

References 39 publications

Recent Advances in Chiral Analysis of Proteins and Peptides

Recent Advances in Chiral Analysis of Proteins and Peptides

Recent advances of the ionic chiral selectors for chiral resolution by chromatography, spectroscopy and electrochemistry

Comprehensive Online Reversed-Phase × Chiral Two-Dimensional Liquid Chromatography-Mass Spectrometry with Data-Independent Sequential Window Acquisition of All Theoretical Fragment-Ion Spectra-Acquisition for Untargeted Enantioselective Amino Acid Analysis

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