What are the most common HPLC modes?

Most common HPLC modes


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

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