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How do I reconcile 98.2% from Janoshik with 97.9% from the supplier?

Asked 9 Jun 2026Modified 1 min agoViewed 2.3k times
8

Numbers first: 98.2% · Janoshik · 97.9%.

I have a result I cannot explain, and I would rather diagnose it than guess.

I have checked the obvious explanations and eliminated the two easiest ones.

What is the differential here, and which test discriminates between the options?

purity
purity

Purity as chromatographic area per cent - the fraction of detected material that is your target peak. It says nothing about how much material is…

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MS
askedmarta_szymanska17k389 Jun 2026
8Do you have a reference for the last claim? Not disputing it, just want to read it. – lane_transit 8 months ago
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5 Answers

Accepted answer first, then by votes
66

Accepted answer

Read the chromatogram before you read the number, because the number without the trace is not a measurement, it is a claim.

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

Column pore size affects mass transfer — a 100 Angstrom packing on a 5 kDa peptide restricts diffusion, broadening the peak and potentially hiding small impurities in the shoulders.

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

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

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TA
answered · acceptedtri_gly_ala48k3817 Jun 2026
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26

In practice, 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.

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.

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.

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

edited 6 Aug 2026 by mz_4113 — expanded the table to cover the lower concentration

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M4
answeredmz_411399k25815 Jul 2026
20

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

Gradient slope controls resolution, and gentler slopes resolve co-eluting impurities into separate peaks — so the better method reports the worse purity number.

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.

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

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DB
answeredDr_Signe_Baldursdottir46k3812 Jun 2026
5Useful. I have added the accept threshold suggestion to my own notes. – tobias_maartens 6 months ago
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16

In practice, the method matters more than the vial, which is why specifying a method buys you far more than changing suppliers does.

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

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 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.

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HL
answeredharriet_lonsdale14k2710 Jul 2026
14

Put another way, start from what the detector sees, because that tells you what the number means.

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.

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

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DF
answeredDr_Nadia_Farsi90k25827 Jun 2026

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