| HPLC mode |
Mobile phase |
Stationary phase |
Analytes |
| Separation of small molecules (up to 2000 Da) |
| Reversed Phase RP (<200 Å) |
Polar (e.g. mixturesfrom water andacetonitrile) |
Nonpolar (e.g. C18) |
Mid-polar - mid-nonpolar soluble in polar and aqueous solvents |
| Normal Phase NP |
Nonpolar (e.g. heptan) |
Polar (e.g. SiOH) |
Nonpolar soluble in nonpolar solvents, insoluble in water |
| Hydrophilic Interaction Liquid Chromatography HILIC |
Polar organic solvents + water |
Polar (e.g. SiOH) |
hydrophilic and highly polar, not retained by RP |
| |
Water layer between stationaryand mobile phase |
|
| Separation of biomolecules (larger than 2000 Da) |
| Wide Pore Reversed phase RP (>200 Å) |
Polar |
Nonpolar |
Mid-polar - mid-nonpolar soluble in polar and aqueous solvents |
| Polymer gel based stationary phases |
| Ion Exclusion & Ligand exchange |
Water, acidic water |
Organic resin with charged groups |
Sugars, organic acids, alcohols |
