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Reversed phase - The most common HPLC mode.
Reversed phase chromatography (RPC) is in theory closely related to hydrophobic interaction chromatography. Both techniques are based upon the interaction between hydrophobic patches on the surface of biomolecules and the hydrophobic groups covalently attached to the surface of the matrix. In practice, however, they are different.
Media for reversed phase chromatography are typically highly substituted with hydrophobic ligands and the binding of substances to reversed phase chromatography media is usually very strong and requires organic solvents for elution. The technique is mainly applicable for peptides, proteins up to Mr 2.5 x 104 and other low molecular weight biomolecules which are stable in aqueous-organic solvents.
Reversed phase chromatography is a widely used analytical technique but it is also required in preparative applications, up to process scale, for very difficult polishing problems like separating microheterogeneities from the native molecule of recombinant peptides. It is also a standard technique for the purification of synthetic peptides and oligonucleotides. Reversed phase chromatography requires medium- to high-pressure equipment, often explosion-proof, for handling the high concentrations of organic solvents.
Read more about the basic principles behind reversed phase chromatography.
Read about our reversed phase chromatography products.
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