Reverse-phase HPLC (revision 1)
Old revision·23:06, 2 Jul 2024·TFA_Counterion
This is an old revision of this page, as it stood at 23:06, 2 Jul 2024, saved by TFA_Counterion with the summary start article on the analytical method. It may differ substantially from the current revision, and any error it contains may since have been corrected.
| Reverse-phase HPLC | |
|---|---|
| Abbreviation | RP-HPLC |
| Stationary phase | Alkyl chains bonded to silica, usually C18 |
| Mobile phase | Water and acetonitrile with an acidic modifier |
| Detection | UV at 214 nm; sometimes 280 nm |
| Analytical method infobox · conventions | |
Reverse-phase HPLC is the mode of liquid chromatography in which the stationary phase is non-polar and the mobile phase is polar — the reverse of the normal-phase arrangement from which the name derives. It is the dominant analytical technique for peptides and the source of almost every purity figure on a peptide certificate.[1]
Separation follows hydrophobicity. A peptide adsorbs to the alkyl stationary phase from a largely aqueous mobile phase and desorbs as the organic proportion rises; the organic content at which it desorbs is characteristic and reproducible under fixed conditions.[2]
References
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