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Preparative HPLC purification (revision 1)

Old revision·09:00, 1 Aug 2024·TesamorelinTeg

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Preparative HPLC purification
ObjectiveCollect a fraction, not measure one
ScaleGrams to kilograms per run
Trade-offYield against purity
Analytical method infobox · conventions

Preparative HPLC purification applies the chromatography of reverse-phase separation at a scale intended to collect material rather than to analyse it. For synthetic peptides it is the step that converts a crude product into a usable one, and it is typically the most expensive stage of manufacture.[1]

The governing trade-off is yield against purity. Collecting a narrow window around the peak apex gives high purity and discards a substantial fraction of the product; collecting more widely recovers more material and includes more of the shoulders where impurities elute. Where the cut is made is a commercial decision as much as a technical one.[2]

References

  1. ^ United States Pharmacopeia, General Chapter <1503>, Quality Attributes of Synthetic Peptide Drug Substances.
  2. ^ Mant CT, Hodges RS. "Analysis of peptides by high-performance liquid chromatography." Methods in Enzymology 271:3–50 (1996). PMID 8782429.