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Analytics · HPLC & UHPLC

Coming back to: Column chemistry choices for a 40-residue peptide

ML
m.lindqvistTL210 Jan 2025#1

On the subject in the title: Column chemistry choices for a 40-residue peptide Working notes rather than a conclusion.

Column chemistry choices — I have the observation and I do not trust my interpretation of it, so I am posting the observation and holding the interpretation back.

Numbers, method and the conditions under which they were collected are below. Interpret them however they warrant.

1 like 19mo
FP
forest_plotTL3Evidence synthesis24 Feb 2025#2

Coming back to the opening post, because the follow-up matters more than the original answer.

Detection wavelength: 214 nm detects the peptide bond and is relatively insensitive to composition. 280 nm detects aromatic residues and is strongly composition-dependent. Area percent at one wavelength is not area percent at the other.

I checked the source rather than the summary, and they differ.

4 likes 17mo
HB
h.bakkerTL227 Mar 2025#3

Reversed-phase separates on hydrophobicity. A peptide is retained on a non-polar stationary phase and eluted by increasing organic solvent. For peptides the mobile phase almost always contains an ion-pairing acid, typically 0.1% TFA, which suppresses secondary interactions and sharpens peaks.

I would put a moderate confidence on that and no more.

13 likes 16mo

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