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Published on: December 4, 2017
Reversed-phase HPLC of peptides: Assessing column and solvent selectivity on standard, polar-embedded and polar
Colin T Mant1, Dziuleta Cepeniene, Robert S Hodges
1Department of Biochemistry and Medical Genetics, University of Colorado Denver, School of Medicine, Anschutz Medical Campus, Aurora, CO 80045, USA.
Abstract:
We desired to evaluate the chromatographic selectivity for peptides of silica-based RP high-performance liquid chromatography stationary phases with various modifications (polar embedding and polar endcapping on C(18) columns; ether-linked phenyl column with polar endcapping) compared with n-alkyl (C(18), C(8)) and aromatic phenylhexyl columns. Thus, we have designed and synthesized two series of synthetic peptide standards with the sequence Gly-Gly-Leu-Gly-Gly-Ala-Leu-Gly-X-Leu-Lys-Lys-amide, where the N-terminal either contains a free α-amino group (AmC series) or is N(α)-acetylated (AcC series) and where position X is substituted by Gly, Ala, Val, Ile, Phe or Tyr. These represent series of peptides with single substitutions of n-alkyl (Gly
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