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Making peptides · Part 5: Purification

How chromatography sorts molecules

Lesson 4.5.1 · 6 min read · Advanced plan

What you'll learn

  • Chromatography separates molecules by how strongly they cling to a packed column as liquid flows through.
  • Reversed-phase HPLC is the most widely used HPLC mode for peptides and sorts them by hydrophobicity.
  • A rising gradient of organic solvent washes peptides off in order of increasing hydrophobicity, and UV detection near 210 nm shows them as peaks.
  • TFA is often added to the liquid, and trifluoroacetate is one of the counter-ions a peptide drug substance can carry.

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