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ForumsCOA & Analytical TestingUnderstanding peptide purity percentages — what worked for you?

Understanding peptide purity percentages — what worked for you?

LondonLisa Wed, Apr 16, 2025 at 10:14 AM 38 replies 2,088 viewsPage 1 of 8
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LondonLisa
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Apr 16, 2025 at 10:14 AM#1

Putting this up for argument rather than for agreement. I have read it twice and I am still not certain what it supports.

Inter-laboratory variation comes from real methodological differences, not sloppiness: which reference standard was used and how it was itself calibrated, the gradient and column, the detection wavelength, and where the integration baseline was drawn. A percentage point or two between competent labs is expected. What is not expected is a large gap, and the usual explanation there is that one report is not of the batch in your hand.

Where I think it is weakest: the comparator does most of the work in how this gets reported, and it is not the comparator most people think they are citing.

What I actually want to know is whether a purity figure and a content figure can disagree without one of them being wrong. Numbers rather than impressions, if you have them.

Note on sourcing:
Figures above are from the primary publication rather than the press summary. If a number here disagrees with one you have, post yours and we will work out which of us is reading a secondary source.
37 7SaraMom3, Dr.MetabolicMD, RetaRick_CA and 34 others
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pete_manc_UK
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Apr 16, 2025 at 11:32 AM#2
LondonLisa said:
Inter-laboratory variation comes from real methodological differences, not sloppiness: which reference standard was used and how it was itself…

Agreeing with LondonLisa, and the qualification matters more than the agreement. They are not contradictory, they measure different things, and this is the single most common misreading of a certificate. Purity is a relative area measurement: the main peak as a percentage of total peak area in the chromatogram. Content is an absolute mass measurement against a reference standard. A vial can be 99% pure and contain 6.1mg — pure material, overfilled — or 99% pure and contain 3mg, which is pure material and a short fill. Purity tells you what fraction of what is there is the right molecule; content tells you how much of it there is. You need both, and a certificate with only purity is telling you half the story.

Last edited: Apr 16, 2025 at 5:32 PM
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PeptideChemSF
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Apr 16, 2025 at 12:50 PM#3
LondonLisa said:
Inter-laboratory variation comes from real methodological differences, not sloppiness: which reference standard was used and how it was itself…

I am more sceptical of certificates as a category than this thread is. The document tells you about a sample somebody chose to send, at a moment in time, and nothing about the vial in your hand. Independent purchase-and-test is a different quality of evidence and everything else is inference.

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TinaHashiRN
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Apr 16, 2025 at 2:08 PM#4

Taking the question as asked, rather than the general version of it. The provenance questions are more useful than the numbers. Does the report name the batch on your vial. Is the date consistent with when that batch existed. Is it addressed to the vendor or to the person who paid for the test. Was the sample taken from general stock or supplied by the vendor. A perfect certificate for a different batch is worth nothing, and vendor-supplied samples are the most common way a genuine report ends up misleading.

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tom_AK
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Apr 16, 2025 at 9:39 PM#5
pete_manc_UK said:
They are not contradictory, they measure different things, and this is the single most common misreading of a certificate.

This is my experience too, for whatever a second data point is worth.

Last edited: Apr 17, 2025 at 1:39 AM
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