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Peptide content (revision 3)

Old revision·17:34, 15 Sep 2024·AnalyticalAnnie

This is an old revision of this page, as it stood at 17:34, 15 Sep 2024, saved by AnalyticalAnnie with the summary clarify that area percent is not mass percent. It may differ substantially from the current revision, and any error it contains may since have been corrected.
Peptide content
Analytical method infobox · conventions

Peptide content, sometimes called peptide net content, is the proportion of the mass of a preparation that is the peptide itself. Everything else in the vial — water taken up during and after lyophilisation, the counterion left by the purification step, residual solvent, and any excipient — is excluded from the numerator and included in the denominator.[1]

Content is the determination that makes a mass statement possible. A purchaser reconstituting a nominally 5 mg vial and calculating a concentration is implicitly assuming a content figure, and in the ordinary case the assumption made is 100%, which is never correct for a lyophilised peptide salt. Typical content for such material falls between roughly 70% and 90%, so the assumption is wrong by an amount that matters.[1]

What occupies the rest of the mass

[edit]

For a lyophilised peptide purified by reverse-phase chromatography and freeze-dried from an aqueous acetonitrile mobile phase, four components account for essentially all of the non-peptide mass.

ComponentTypical shareDetermined by
Water3–10%Karl Fischer titration or Loss on drying
Counterion (usually trifluoroacetate)5–20%Ion chromatography; inferred from the synthesis route only as a last resort
Residual solventbelow 1%Headspace gas chromatography
Peptide-related impurities0.5–3%The complement of Area percent purity

References

  1. ^ a b United States Pharmacopeia, General Chapter <1503>, "Quality Attributes of Synthetic Peptide Drug Substances" (informational). USP–NF, current revision. Sets out peptide content as an attribute distinct from chromatographic purity.