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Analytical methods

HPLC and mass spectrometry: what each test actually answers

Two tests appear on almost every peptide certificate. They answer different questions, and reading one as though it answered the other is the most common misreading of the document.

6 min readResearch operationsUpdated 18 Aug 2026

Purity is a proportion, not an identity

High performance liquid chromatography separates a sample into its components and measures how much of the total each one represents. The headline percentage is the share of the sample made up by the largest peak.

That tells you how much of the material is one substance. On its own it does not tell you which substance that is. A very pure sample of the wrong compound still reports as very pure.

Identity is a question of mass

Mass spectrometry measures the mass to charge ratio of the molecules present, which acts as a fingerprint. Comparing the measured mass against the expected mass for a known sequence is what confirms the material is what the label says.

This is why certificates report identity as confirmed or not confirmed rather than as a percentage. It is a yes or no question about which molecule is present.

Reading the pair together

Taken together the two answer: this is the compound we expected, and it makes up this share of the sample. Either result alone leaves an obvious gap.

A certificate that reports only a purity figure is incomplete in a specific way, and worth asking about. So is one that confirms identity but never quantifies the material.

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