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Preparative HPLC System

TL;DR

Preparative HPLC systems purify peptides at 50 mg to 10+ g per run. Key specifications include pump flow rate (up to 150 mL/min), column dimensions, and UV detection at 214/254 nm.

Category

Purification Equipment

Application

Purification of crude peptides to target purity (typically ≥98%)

Function

Remove deletion sequences, truncated peptides, and organic impurities from crude SPPS product.

System Parameters

Parameter Research Scale Process Scale
Column ID 10–30 mm 50–200 mm
Max flow rate 5–50 mL/min 100–1000 mL/min
Max pressure 200–400 bar 100–200 bar
Injection volume 0.1–10 mL 10–200 mL
Detection UV 214, 254, 280 nm UV 214, 254, 280 nm
Fraction collector Time or peak-based Peak-based

Column Specifications

Property Typical
Stationary phase C18 silica
Particle size 5–10 μm (research), 10–30 μm (process)
Pore size 100–300 Å
Column length 150–300 mm

Mobile Phase

Phase Composition
A 0.05–0.1% TFA in water
B 0.05–0.1% TFA in acetonitrile
  • Peptide purification
  • Fraction screening by analytical HPLC

  • Pooling of high-purity fractions

  • Large-Scale Peptide Purification — Review the section above for key parameters, methods, and quality criteria.

  • System Parameters — Review the section above for key parameters, methods, and quality criteria.
  • Column Specifications — Review the section above for key parameters, methods, and quality criteria.

🔗 Related: Purification Process | Analytical HPLC | HPLC Analysis | Peptide Quality Control Guide

Key Takeaways

  • Application — Review the section above for key parameters, methods, and quality criteria.
  • System Parameters — Review the section above for key parameters, methods, and quality criteria.
  • Column Specifications — Review the section above for key parameters, methods, and quality criteria.
  • Mobile Phase — Review the section above for key parameters, methods, and quality criteria.