title: Separation of peptides on superficially porous particle based macrocyclic glycopeptide liquid chromatography stationary phases: consideration of fast separations.
abstract: Macrocyclic glycopeptide based liquid chromatography stationary phases are known for their highly selective peptide separations. Fast and ultrafast (tR< 1 min) high-efficiency separations were achieved with superficially porous particle (SPP)-based stationary phases. Separations of pharmaceutically important classes of peptides such as enkephalins and bradykinins have been achieved in less than 5 min in isocratic elution modes. Selectivity for peptides structurally similar to one another was increased with use of teicoplanin-based stationary phases compared with commercial C18stationary phases. Ultrafast isocratic separations of structurally related peptides were achieved with teicoplanin- and vancomycin-based short SPP columns. Acidic mobile phases produced better separations. Ammonium formate was the optimal mobile phase buffer additive. Use of an appropriate combination of a macrocyclic glycopeptide stationary phase and a mobile phase permits faster and more electrospray ionization mass spectrometry compatible isocratic separations than previous gradient approaches. The tryptic peptide separation characteristics of the teicoplanin stationary phase are demonstrated. Additionally, compared with commercial C18stationary phases, teicoplanin showed tryptic peptide separations with different selectivities. Graphical Abstract Ultrafast separation of enkephalin peptide epimers.
authors.affiliation: Department of Chemistry and Biochemistry, The University of Texas at Arlington, Arlington, TX, 76019, USA.
authors.forename: Rasangi M
authors.initials: RM
authors.lastname: Wimalasinghe
authors.affiliation: Department of Chemistry and Biochemistry, The University of Texas at Arlington, Arlington, TX, 76019, USA.
authors.forename: Zachary S
authors.initials: ZS
authors.lastname: Breitbach
authors.affiliation: AZYP LLC, 700 Planetarium Place, Arlington, TX, 76019, USA.
authors.forename: Jauh T
authors.initials: JT
authors.lastname: Lee
authors.affiliation: Department of Chemistry and Biochemistry, The University of Texas at Arlington, Arlington, TX, 76019, USA. sec4dwa@uta.edu.|AZYP LLC, 700 Planetarium Place, Arlington, TX, 76019, USA. sec4dwa@uta.edu.
authors.forename: Daniel W
authors.initials: DW
authors.lastname: Armstrong
mesh_terms: D000595:Amino Acid Sequence; D006020:Glycopeptides / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D047028:Macrocyclic Compounds / Q000737:chemistry*; D010455:Peptides / Q000302:isolation & purification*; D016062:Porosity; D013237:Stereoisomerism
chemical_list: D006020:Glycopeptides; D047028:Macrocyclic Compounds; D010455:Peptides
keywords: Fast separations; Peptides; Teicoplanin; Tryptic peptides; Ultrafast separations; Vancomycin