Peptide Reversed-Phase Columns

Reversed-Phase Chromatography

Reversed-phase chromatography using ion-pairing reagents such as TFA and formic acid can deliver highly resolved separations of complex peptide mixtures, (e.g., tryptic protein digests or lengthy synthetic peptide sequences) whose sequences may differ by a single amino acid. In general, the hydrophobicity of the peptide determines the elution order, with the least hydrophobic peptides eluting first.  

Factors such as particle composition (silica vs hybrids), pore size (130 Å vs 300 Å), ligand density, as well as separation conditions (e.g., gradient duration, separation temperature, flow rate) all play an important role in obtaining a separation that meets application requirements.

As with other bioseparations, when using smaller particle sizes, UPLC/UHPLC column offerings provide superior component resolution and in less time compared to HPLC-based columns for this demanding application.

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