How do I reconcile 96.8% from Medutest with 98.8% from the supplier?

Setup, so nobody has to ask: 96.8% · Medutest · 98.8%.

This is not behaving the way I expected and I want to understand the discrepancy before I act on it.

I have photographed the current state and recorded the conditions, so I can answer follow-up questions precisely.

How do I distinguish the benign explanation from the one that matters?

Mechanically, identity and purity are orthogonal, and a high purity says almost nothing about whether the peak is actually what you think it is.

Mass on column affects detector linearity and peak overlap — overloading broadens peaks and hides neighbours, while underloading improves resolution but loses sensitivity.

Retention time is sequence-specific and method-specific, so comparing your result to a supplier value using a different method is meaningless without method documentation.

Proline conformer interconversion kinetics are well-characterised and the half-life is of the same order as the chromatographic peak width at room temperature.

The limitation is that single-digit micro-impurities become invisible at typical reporting thresholds, so "no impurities detected" means "none above one in two thousand."

The practical summary: ask for the chromatogram and the method, and ignore the headline number until you have both.

On the detail: understanding purity requires separating the chemistry from the method from the reporting convention, and the three are not independent.

Temperature affects the dynamics of molecular conformation, and if a peptide has proline residues that interconvert on the chromatographic timescale, the peak will split or shoulder at low temperature and collapse at high temperature.

Specifically, the fraction of your main peak that is actually your target versus isomers, fragments or related sequences is invisible without complementary identity testing.

The ICH Q3A impurity thresholds and the relevant pharmacopoeial chapters all specify method validation requirements that almost no research-grade certificate claims to meet.

If you are ranking vendors, specify a method and have all samples tested at the same place.

edited 12 Sept 2025 by tandem_gradient — added a caveat about sampling

Mechanically, the honest answer is that the achievable range of plausible purity figures for a given vial is wider than most people expect.

Mobile phase additive choice affects ionisation and peak shape — TFA gives sharp peaks but suppresses mass spectrometry signal, formic acid gives worse peaks but preserves signal.

Inter-laboratory studies on identical peptide material routinely find half-a-per-cent to a full-per-cent spreads in reported purity on the same sample.

Compare purity within a single laboratory on the same method, never across laboratories.

Gradient slope is the most powerful parameter and almost nobody mentions it, which is why two reports on the same material disagree by a point.

Buffer versus acid in the mobile phase changes the ionisation state of basic and acidic residues, shifting retention and selectivity — same vial, potentially different separation.

The Arrhenius relationship for peptide degradation is the basis of accelerated stability testing and also governs how quickly methods drift with temperature.

The caveat is that purity without identity is only half an answer — a high purity does not mean the peak is actually what you think it is.

The practical summary: ask for the chromatogram and the method, and ignore the headline number until you have both.

edited 27 Sept 2025 by mira_sundqvist — expanded the table to cover the lower concentration