Reverse-phase HPLC: difference between revisions
Diff·revision 5 → 6·14:36, 12 Aug 2024
Difference between revision 5 and revision 6 of Reverse-phase HPLC. 3 lines changed; the page grew by 439 bytes.
| Revision 5 — 17:42, 6 Aug 2024 FormularyFrida (talk) state the column chemistry and the gradient the figure was obtained on 2,305 bytes ±0 | Revision 6 — 14:36, 12 Aug 2024 TFA_Counterion (talk) add short description 2,744 bytes +439 | ||
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| 11 | 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.{{r|mant1996}} | 11 | 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.{{r|mant1996}} |
| 12 | 12 | ||
| + | 13 | An acidic modifier — most often trifluoroacetic acid — is added to suppress ionisation of acidic side chains and to pair with basic ones, sharpening peaks considerably. Its use is why peptides purified by this route emerge as [[Trifluoroacetate counterion|trifluoroacetate salts]].{{r|usp1503}} | |
| + | 14 | ||
| 13 | == Conditions and what they control == | 15 | == Conditions and what they control == |
| 14 | | Parameter | Typical value | What it changes | | 16 | | Parameter | Typical value | What it changes | |
| ⋮ | ⋮ | ||
| 28 | <ref name="usp621">United States Pharmacopeia, General Chapter <621>, ''Chromatography''.</ref> | 30 | <ref name="usp621">United States Pharmacopeia, General Chapter <621>, ''Chromatography''.</ref> |
| 29 | <ref name="mant1996">Mant CT, Hodges RS. "Analysis of peptides by high-performance liquid chromatography." ''Methods in Enzymology'' 271:3–50 (1996). PMID 8782429.</ref> | 31 | <ref name="mant1996">Mant CT, Hodges RS. "Analysis of peptides by high-performance liquid chromatography." ''Methods in Enzymology'' 271:3–50 (1996). PMID 8782429.</ref> |
| + | 32 | <ref name="usp1503">United States Pharmacopeia, General Chapter <1503>, ''Quality Attributes of Synthetic Peptide Drug Substances''.</ref> | |
| 30 | 33 | ||
| 31 | {{DEFAULTSORT:Reverse-phase HPLC}} | 34 | {{DEFAULTSORT:Reverse-phase HPLC}} |