HPLC or high performance liquid chromatography is the heart of pharmaceutical analysis. Where high pressure is used to pass the target molecule with mobile phase throgh the compacted stationary phase. In this article we mention 10 tips for mobile phase of HPLC.
1. Choose the right solvent
The choice of solvent for the mobile phase is critical in HPLC. It should be compatible with the sample and stationary phase, and have a low UV cut off to avoid interference with the detector.
2. Optimize pH
The pH of the mobile phase can affect the retention time and selectivity of analytes. Adjusting the pH can improve separation efficiency and peak shape.
3. Use a UV cut-off filter
A UV cut-off filter can be used to remove any unwanted UV light from the mobile phase, which can interfere with detection.
4. Avoid contamination
Contamination of the mobile phase can lead to poor separation and inaccurate results. Use high-quality solvents and filters to prevent contamination.
5. Monitor column performance
Regular monitoring of column performance is essential to ensure optimal separation efficiency and peak shape.
6. Use gradient elution
Gradient elution can improve separation efficiency by allowing for better resolution of complex mixtures.
7. Optimize flow rate
The flow rate of the mobile phase affects retention time, resolution, and peak shape. Optimizing flow rate can improve separation efficiency.
8. Use appropriate buffers
Buffers are used to adjust pH in HPLC mobile phases, but they can also affect retention time and selectivity. Choose appropriate buffers that are compatible with your sample and stationary phase.
9. Check for interference
Some components in the mobile phase may interfere with detection or cause baseline drifts. Check for interference regularly to ensure accurate results.
10. Store solvents properly
Proper storage of solvents is critical to prevent degradation or contamination that could affect HPLC performance or results. Store solvents in a cool, dry place away from direct sunlight or heat sources.
Read also:
- Pharmaceutical Quality Control Equipment List
- User Requirement Specification for HPLC
- Basic Knowledge on Chromatographic Technique
