Reverse-phase HPLC: difference between revisions
Diff·revision 11 → 12·09:53, 19 Nov 2024
Difference between revision 11 and revision 12 of Reverse-phase HPLC. 6 lines changed; the page grew by 697 bytes.
| Revision 11 — 10:17, 1 Nov 2024 NotabilityNoa (talk) give the limit of quantitation with the signal-to-noise ratio it assumes 3,045 bytes ±0 | Revision 12 — 09:53, 19 Nov 2024 TechnicianTeal (talk) correct the resolution formula — the denominator is the sum of the widths 3,742 bytes +697 | ||
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| 1 | {{Infobox method | 1 | {{Infobox method |
| 2 | | name = Reverse-phase HPLC | 2 | | name = Reverse-phase HPLC |
| + | 3 | | subtitle = Chromatographic mode | |
| 3 | | image = chromatogram.svg | 4 | | image = chromatogram.svg |
| 4 | | Abbreviation = RP-HPLC | 5 | | Abbreviation = RP-HPLC |
| ⋮ | ⋮ | ||
| 28 | 29 | ||
| 29 | Temperature is the parameter most often omitted from certificates. Elevated column temperature sharpens peaks for large peptides by reducing conformational heterogeneity, and a method developed at 60 °C will not reproduce at ambient. | 30 | Temperature is the parameter most often omitted from certificates. Elevated column temperature sharpens peaks for large peptides by reducing conformational heterogeneity, and a method developed at 60 °C will not reproduce at ambient. |
| + | 31 | ||
| + | 32 | == Why it suits peptides == | |
| + | 33 | Peptides are amphipathic and their hydrophobicity varies with sequence, so retention differences between closely related species are often large enough to separate them. A single-residue deletion changes hydrophobicity measurably in most cases, which is why the technique can resolve synthesis impurities that differ very little in mass.{{r|mant1996}} | |
| + | 34 | ||
| + | 35 | The technique is also tolerant. Samples can be injected in aqueous solution, the mobile phases are simple and volatile enough to be compatible with [[Mass spectrometry|mass spectrometry]], and the same chemistry scales to [[Preparative HPLC purification|preparative]] loadings. | |
| 30 | 36 | ||
| 31 | == References == | 37 | == References == |