High-performance liquid chromatography: difference between revisions
Diff·revision 3 → 4·03:46, 10 Jul 2024
Difference between revision 3 and revision 4 of High-performance liquid chromatography. 3 lines changed; the page grew by 491 bytes.
| Revision 3 — 11:22, 29 Jun 2024 Retention_Time_Rae (talk) typo 2,129 bytes ±0 | Revision 4 — 03:46, 10 Jul 2024 Retention_Time_Rae (talk) state the column chemistry and the gradient the figure was obtained on 2,620 bytes +491 | ||
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| 15 | In reverse-phase separation the stationary phase is a hydrocarbon chain, commonly C18, bonded to silica particles. Hydrophobic molecules are retained more strongly. Increasing the proportion of organic solvent in the mobile phase weakens that retention, so a gradient of increasing organic content elutes components in approximate order of hydrophobicity. | 15 | In reverse-phase separation the stationary phase is a hydrocarbon chain, commonly C18, bonded to silica particles. Hydrophobic molecules are retained more strongly. Increasing the proportion of organic solvent in the mobile phase weakens that retention, so a gradient of increasing organic content elutes components in approximate order of hydrophobicity. |
| 16 | 16 | ||
| + | 17 | Resolution between two peaks depends on their retention difference, on peak width, and on the efficiency of the column. Efficiency improves with smaller particles and with longer columns, at the cost of higher back-pressure — the constraint that drove the development of sub-2-micron particles and the instruments capable of running them.{{r|snyder2010}} | |
| + | 18 | ||
| 17 | == References == | 19 | == References == |
| 18 | {{reflist}} | 20 | {{reflist}} |
| 19 | <ref name="usp621">United States Pharmacopeia, General Chapter <621>, ''Chromatography''.</ref> | 21 | <ref name="usp621">United States Pharmacopeia, General Chapter <621>, ''Chromatography''.</ref> |
| 20 | <ref name="usp1503">United States Pharmacopeia, General Chapter <1503>, ''Quality Attributes of Synthetic Peptide Drug Substances''.</ref> | 22 | <ref name="usp1503">United States Pharmacopeia, General Chapter <1503>, ''Quality Attributes of Synthetic Peptide Drug Substances''.</ref> |
| + | 23 | <ref name="snyder2010">Snyder LR, Kirkland JJ, Dolan JW. ''Introduction to Modern Liquid Chromatography'', 3rd edition (2010).</ref> | |
| 21 | 24 | ||
| 22 | {{DEFAULTSORT:High-performance liquid chromatography}} | 25 | {{DEFAULTSORT:High-performance liquid chromatography}} |