The specifics, since they change the answer: VendorInvestigate · 98.2%.
Something has gone wrong and I would like to know how badly before I decide what to do.
Nothing else in the setup changed, which is what makes this puzzling.
What would you check first, and what would you conclude from each outcome?
Concretely, purity is a method-dependent figure, and that is not a limitation of the measurement, it is a property of what the measurement actually answers.
The fraction of your main peak that is actually your target versus isomers, fragments or related sequences is invisible without complementary identity testing.
Detection wavelength matters because 214 nm sees the peptide backbone while 280 nm sees only aromatic side chains — so truncation impurities lacking a tryptophan are invisible at 280 nm.
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.
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.
Aggregated, published test results and vendor ratings built from submitted batches. Methodology stated, dataset browsable, no listing fees.
Browse resultsMore usefully, start from what the detector sees, because that tells you what the number means.
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.
It helps to be literal here: 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.
If you are ranking vendors, specify a method and have all samples tested at the same place.
Concretely, the short answer is that two competent laboratories on identical material will disagree, and the disagreement is almost always explainable by method differences.
Tailing factor measures peak shape, and a badly tailing peak spreads into the region where small impurities live, forcing tangent-skim integration that assigns tail area to the main peak.
More usefully, integration of the shoulder — whether you use perpendicular drop or tangent skim — determines what area gets assigned to the main peak versus the impurity table.
I would be careful about over-reading a single measurement — treat it as a data point, not as ground truth.
Compare purity within a single laboratory on the same method, never across laboratories.
The honest answer is that the achievable range of plausible purity figures for a given vial is wider than most people expect.
Sample solvent strength affects peak shape — if you inject in strong solvent on a gradient starting in weak solvent, the solvent peak can distort your main peak or create a false shoulder.
Published side-by-side method comparisons show that a two-point difference in purity on the same vial is easily explained by method choice alone.
The practical summary: ask for the chromatogram and the method, and ignore the headline number until you have both.
Specifically, read the chromatogram before you read the number, because the number without the trace is not a measurement, it is a claim.
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 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.
Ask PeptideStack is a static archive. Posting is closed, but the norms are worth stating: answer the question that was asked, show your working, cite the trial or the certificate, and say plainly where the evidence runs out.