Reversed-phase high-performance liquid chromatography separates a sample across a stationary phase; the resulting chromatogram expresses relative abundance as peak area. A single dominant peak at the expected retention time, with a clean baseline, is the visual signature of a well-purified preparation.
Mass spectrometry does something chromatography cannot: it weighs the molecule. Electrospray ionisation of a peptide typically yields a family of multiply charged ions that deconvolute to one monoisotopic mass, which is then compared against the theoretical mass of the stated sequence.
Neither instrument is sufficient alone. Purity without identity describes a very pure unknown; identity without purity describes the right molecule in unknown company.
Research Status
Established analytical chemistry rather than an evolving evidence base -- the open questions here are about method validation, not biological effect.
Selected Research
- NIST. Mass Spectrometry Data Center. National Institute of Standards and Technology, 2024.
- USP. General Chapter <621> Chromatography. United States Pharmacopeia, 2024.
Related Research