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Karl Fischer titration: difference between revisions

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Revision 7 — 01:19, 21 Jul 2024
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10The method matters to readers of a [[Certificate of analysis|certificate of analysis]] for a specific and frequently misunderstood reason: a chromatographic purity figure is a statement about the ''relative'' composition of the material that dissolved, not about ''how much'' peptide the vial contains. Water, counterions and residual salts occupy mass. A vial of material reported at 99% purity by [[High-performance liquid chromatography|HPLC]] may nevertheless be 6% water and 8% [[Trifluoroacetate counterion|trifluoroacetate]] by mass, in which case its [[Peptide content|peptide content]] is closer to 85%.{{r|usp1503}}10The method matters to readers of a [[Certificate of analysis|certificate of analysis]] for a specific and frequently misunderstood reason: a chromatographic purity figure is a statement about the ''relative'' composition of the material that dissolved, not about ''how much'' peptide the vial contains. Water, counterions and residual salts occupy mass. A vial of material reported at 99% purity by [[High-performance liquid chromatography|HPLC]] may nevertheless be 6% water and 8% [[Trifluoroacetate counterion|trifluoroacetate]] by mass, in which case its [[Peptide content|peptide content]] is closer to 85%.{{r|usp1503}}
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+12== Chemical basis ==
+13The underlying reaction is the oxidation of sulfur dioxide by iodine, which consumes water stoichiometrically:
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+15{{math|H_{2}O + I_{2} + SO_{2} + CH_{3}OH + 2\,RN → 2\,RNH^{+}I^{-} + RNH^{+}CH_{3}SO_{4}^{-}}}
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+17Methanol participates directly, forming a methyl sulfite intermediate that is the species actually oxidised; the base (historically pyridine, now typically imidazole) buffers the liberated acid and keeps the reaction in the pH window where the stoichiometry holds at 1 mol iodine per mol water.{{r|scholz1984}}
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12== References ==19== References ==
13{{reflist}}20{{reflist}}
14<ref name="usp921">United States Pharmacopeia, General Chapter <921>, "Water Determination". USP–NF, current revision.</ref>21<ref name="usp921">United States Pharmacopeia, General Chapter <921>, "Water Determination". USP–NF, current revision.</ref>
15<ref name="usp1503">United States Pharmacopeia, General Chapter <1503>, "Quality Attributes of Synthetic Peptide Drug Substances" (informational). USP–NF, current revision.</ref>22<ref name="usp1503">United States Pharmacopeia, General Chapter <1503>, "Quality Attributes of Synthetic Peptide Drug Substances" (informational). USP–NF, current revision.</ref>
+23<ref name="scholz1984">Scholz E. ''Karl Fischer Titration: Determination of Water''. Springer, Berlin (1984). The standard monograph on reagent chemistry and interferences.</ref>
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17{{DEFAULTSORT:Karl Fischer titration}}25{{DEFAULTSORT:Karl Fischer titration}}
18[[Category:Analytical methods]]26[[Category:Analytical methods]]
19[[Category:Compendial testing]]27[[Category:Compendial testing]]
+28[[Category:Certificates of analysis]]
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