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Is a 97.9% result from Janoshik comparable to 99.1% from Medutest?

Asked 19 Mar 2024Modified 2.1 years agoViewed 25k times
6

Details up front: 97.9% · Janoshik · 99.1% · Medutest.

I have used one of these for a while and I am considering switching, which requires a reason.

What I care about is reproducibility, because a result I cannot repeat is not useful to me.

Which axes does this decision turn on?

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TW
askedtare_and_weigh12k1619 Mar 2024
Which wavelength was the purity integrated at? Worth adding to the question. – h_pergande 9 months ago
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5 Answers

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55

97.9 and 99.1 are 1.2 percentage points apart, which sounds small until you restate it as impurity. Janoshik is leaving 2.1 per cent of the detected area unassigned and Medutest 0.9 per cent — a factor of 2.33 between them. The impurity fraction is the quantity that moves when a method changes, and it is the one worth arguing about; the headline is just its complement. Comparable means same column chemistry, same gradient slope, same detection wavelength, same integration convention. Until you have those four from both laboratories, 97.9 and 99.1 are two measurements of slightly different quantities that happen to share a unit.

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.

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

In practice, 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.

I would be careful about over-reading a single measurement — treat it as a data point, not as ground truth.

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

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answeredanouk_desmet16k389 Jul 2024
3Same experience here, different supplier. – Dr_Fatima_Belkacem 2 months ago
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38

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.

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.

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.

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

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RC
answeredRP_C18105k34828 Jun 2024
7I would gently push back on the second point — inter-laboratory spread is wider than stated. – ruaidhri_o_shea 9 months ago
6Do you have the chromatogram for this, or just the summary figure? – kwn_analytical 7 months ago
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30

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.

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

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

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

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SK
answereds_kalniete57k382 Apr 2024
25

The short answer is that two competent laboratories on identical material will disagree, and the disagreement is almost always explainable by method differences.

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

If you only pay for one test, pay for quantified content. Purity is the number everyone quotes and content is the number that changes what you do.

edited 28 Mar 2024 by tobias_reint — fixed an arithmetic slip in the third paragraph

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TR
answeredtobias_reint20k3822 Mar 2024
7Confirming from the other direction: I ignored the method section once and paid for it. – e_dziedzic 8 months ago
6The system-suitability data is the part that tells you whether to believe the rest. – Dr_Jonas_Halvorsen 6 months ago
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17

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

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.

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.

Worth noting that method standardisation is poor in the research-grade space compared to pharmaceutical work, so identical-looking methods can produce different results.

In practice: ask for the chromatogram, check the method section, check the lot number against the vial, and set your accept threshold before you see the result rather than after.

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IB
answeredines_brandt113k25725 Apr 2024
5The impurity table is the part I now read first, and this explains why. – plate_count_9k 8 months ago
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