HILIC Peptide Purification

Alternative selectivity for highly polar peptides and difficult RP retention (GLP / ISO 13485 aligned, non-GMP).

HILIC retains polar peptides and can provide the separation you cannot get with RP-HPLC alone. It is commonly used as an orthogonal step, followed by RP polishing when needed.

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When to use Why it works Typical workflow Deliverables Related pages

When HILIC is the right choice

Best fit

• Highly polar peptides with weak RP retention
• Impurities that resist RP selectivity
• Orthogonal workflows where RP alone is insufficient

Common outcomes

• Improved retention and peak shape for polar sequences
• Different selectivity profile vs RP
• Cleaner fractions prior to RP “polish”

Why HILIC can succeed when RP fails

RP-HPLC is dominated by hydrophobic interactions. Highly polar peptides may show poor retention or compressed peak spacing. HILIC introduces different retention behavior, often creating separation between target and impurities that are “too similar” in RP.

Typical workflow: HILIC → RP-HPLC

• Screen: selectivity scouting to confirm retention and impurity resolution
• Capture: preparative HILIC fractionation
• Polish: RP-HPLC for final purity and consistency (when required)
• Verify: analytical HPLC + MS identity verification (as scoped)

Deliverables

• Analytical HPLC chromatograms + purity assessment
• MS identity verification (project-appropriate)
• COA + supporting data package

Non-GMP: GLP / ISO 13485 aligned workflows. GMP manufacturing is not offered.

Related purification pages

RP-HPLC Ion-Exchange (IEX) Desalting / SEC COA & QC Purification Overview