Peptide analytical testing: HPLC, LC–MS and method limits
Analytical methods answer defined questions about a specified sample. No single result automatically establishes identity, purity, quantity, sterility and suitability for use.
By Manchester Peptides · ·
Chromatography separates components
High-performance liquid chromatography (HPLC) separates components as they pass through a column. A chromatogram can show peaks associated with detected components under the stated conditions. An area-percentage result may describe relative detector response; it should not automatically be read as confirmation of identity, total content or absence of every possible contaminant.
Mass spectrometry measures mass-to-charge
Mass spectrometry can provide evidence about molecular mass and, depending on the method, structural information. Liquid chromatography–mass spectrometry (LC–MS) combines separation with mass analysis. Matching an expected mass can support an identity assessment, but interpretation depends on the method, reference information, sample preparation and the possibility of related compounds.
Method validation matters
An analytical result is only as meaningful as the method’s intended purpose and demonstrated performance. The ICH Q2(R2) framework published by the FDA describes principles for validating analytical procedures. Depending on purpose, characteristics may include specificity, accuracy, precision, range and detection or quantitation capability.
What to look for on a report
- A unique report and sample or batch identifier.
- The method used for each reported result.
- Units, specifications, acceptance criteria and actual observations.
- Dates, laboratory identity and authorisation.
- Clear separation between measured results and interpretation.
Source and related guide
FDA: ICH Q2(R2) Validation of Analytical Procedures. This regulatory guidance is a general analytical framework and does not validate any material shown on this website.
Related: how to read a certificate of analysis →