High-Performance Liquid Chromatography (HPLC) is the gold standard for peptide purity analysis. This comprehensive guide explains how HPLC testing works and why it's essential for verifying research peptide quality.
What is HPLC?
HPLC (High-Performance Liquid Chromatography) is an analytical technique that separates, identifies, and quantifies components in a mixture. For peptides, HPLC is the most reliable method to determine purity and detect impurities.
How HPLC Testing Works
The HPLC Process:
- Sample Preparation: The peptide is dissolved in a suitable solvent
- Injection: The sample is injected into the HPLC system
- Separation: Components pass through a column filled with stationary phase material
- Detection: A detector measures each component as it exits the column
- Analysis: Results are displayed as a chromatogram showing peaks for each component
Key Components of HPLC:
- Mobile Phase: Liquid solvent that carries the sample through the system
- Stationary Phase: Column packing material that separates components
- Detector: UV detector that measures peptide concentration
- Pump: Delivers mobile phase at high pressure
Types of HPLC for Peptides
Reverse-Phase HPLC (RP-HPLC)
The most common method for peptide analysis.
- Uses a hydrophobic stationary phase
- Separates peptides based on hydrophobicity
- Ideal for most research peptides
- Provides excellent resolution
Ion-Exchange HPLC
Separates peptides based on charge.
- Useful for charged peptides
- Complements RP-HPLC analysis
- Detects charge-based impurities
Size-Exclusion HPLC
Separates based on molecular size.
- Identifies aggregates
- Detects degradation products
- Useful for quality control
Reading an HPLC Chromatogram
Understanding your HPLC results is crucial for quality verification.
What to Look For:
- Main Peak: Represents your target peptide - should be tall and sharp
- Peak Area: Indicates peptide quantity - larger area = higher concentration
- Retention Time: Time for peptide to pass through column - should match expected value
- Additional Peaks: Indicate impurities - should be minimal or absent
Purity Calculation:
Purity percentage is calculated by dividing the area of the main peak by the total area of all peaks:
Purity (%) = (Main Peak Area / Total Peak Area) × 100
Quality Indicators:
- ≥98% Purity: Single dominant peak with minimal impurities - research grade
- 95-98% Purity: Main peak with small impurity peaks - acceptable for screening
- <95% Purity: Multiple significant peaks - not suitable for critical research
HPLC vs Other Testing Methods
| Method | Advantages | Limitations |
|---|---|---|
| HPLC | Highly accurate, quantitative, industry standard | Requires specialized equipment |
| Mass Spectrometry | Confirms molecular weight, detects modifications | Doesn't quantify purity as precisely |
| TLC | Quick and inexpensive | Less accurate, qualitative only |
| Amino Acid Analysis | Confirms sequence | Doesn't measure purity percentage |
Why HPLC Matters for Research
HPLC testing ensures:
- Accurate Dosing: Know exactly how much active peptide you're using
- Reproducible Results: Consistent purity leads to reliable experiments
- Impurity Detection: Identify contaminants that could affect results
- Quality Assurance: Verify supplier claims with objective data
- Regulatory Compliance: Meet quality standards for research protocols
Bio Peptides UK HPLC Standards
All our research peptides undergo rigorous HPLC testing:
- RP-HPLC Analysis: Every batch tested using reverse-phase HPLC
- ≥98% Purity Guarantee: Minimum purity standard for all products
- Full Chromatograms: Complete HPLC data included in COA
- Batch Traceability: Every vial linked to specific HPLC results
- Third-Party Verification: Independent laboratory confirmation
Learn more about how peptide purity is tested and our comprehensive quality standards.
Understanding Your COA
Your Certificate of Analysis includes HPLC data:
- Chromatogram Image: Visual representation of purity
- Purity Percentage: Calculated from peak areas
- Retention Time: Confirms peptide identity
- Testing Conditions: Column type, mobile phase, detection wavelength
Learn how to read and verify your COA for complete quality assurance.
Related Resources
Explore our comprehensive guides:
Order HPLC-Tested Research Peptides
All Bio Peptides UK products include full HPLC analysis and COA documentation. Browse our complete range of research peptides.
All products are for research use only.