Reversed-Phase Chromatography & HPLC Columns
Reverse phase chromatography is the most widely used mode of HPLC, ideal for separating a broad range of compounds from pharmaceuticals to biomolecules. In reversed-phase HPLC, the stationary phase is made of hydrophobic, silane-bonded silica, while the mobile phase is typically a polar solvent such as water mixed with organic modifiers. This setup separates analytes based on hydrophobic interactions, with more non-polar compounds eluting later. Adjusting the mobile phase composition allows fine-tuning for optimal resolution.
How Reverse Phase Chromatography Works
In reverse phase HPLC, non-polar stationary phases (e.g., C18, C8, phenyl) interact with analytes based on their hydrophobicity. More hydrophobic analytes spend longer bound to the stationary phase, resulting in later elution times. Mobile phase gradients, pH adjustments, and organic solvent concentrations are used to optimize retention and resolution.
Reverse Phase vs. Normal Phase Chromatography
While reverse phase chromatography uses a non-polar stationary phase and polar mobile phase, normal phase chromatography uses the opposite — a polar stationary phase and non-polar mobile phase. Reverse phase is more common due to its compatibility with aqueous mobile phases and a wider range of analytes.
Orochem Reversed-Phase HPLC Columns
Our reversed-phase HPLC columns are available in multiple chemistries (C18, C8, phenyl, and more), particle sizes, and dimensions to suit both analytical and preparative workflows. Proprietary silica manufacturing ensures:
- Narrow particle size distribution
- High mechanical stability
- Consistent batch-to-batch performance
Order Reverse Phase HPLC Columns
Browse our range of reversed-phase chromatography columns online or contact our technical experts for phase selection and method optimization.
Reversed-Phase
