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

Understanding peptide purity percentages — what worked for you?

andrew_nyc Tue, Jul 9, 2024 at 12:22 PM 25 replies 2,441 viewsPage 1 of 5
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andrew_nyc
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Jul 9, 2024 at 12:22 PM#1

Posting this because the summary going around does not say what the paper says, and the difference matters for how people here are using 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.

Where I think it is weakest: the population was selected and supported in ways a real cohort is not, so I would read the effect size as a ceiling rather than an expectation.

What I am after is why two laboratories report different purity for the same batch, and which number to believe. Tell me what I have not thought of.

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.
14 9Dr.KarenChen, Dr.NateNeph, PharmD_Rodriguez and 11 others
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pete_manc_UK
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Jul 9, 2024 at 12:46 PM#2
andrew_nyc said:
The provenance questions are more useful than the numbers.

Agreeing with andrew_nyc, and the qualification matters more than the agreement. 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.

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SarahChen_PharmD
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Jul 9, 2024 at 1:10 PM#3
andrew_nyc said:
The provenance questions are more useful than the numbers.

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.

12 7paul_denver, TinaHashiRN, robert_kc and 9 others
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Dr.SleepRoch
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Jul 9, 2024 at 1:34 PM#4

This one has a reasonably settled answer, so here it is. 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: Jul 9, 2024 at 5:34 PM
11 6tony_orlando, Dr.NephBHM_UK, kim_atl_prep and 8 others
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hans_munich
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Jul 9, 2024 at 3:44 PM#5
pete_manc_UK said:
Inter-laboratory variation comes from real methodological differences, not sloppiness: which reference standard was used and how it was itself…

Same experience, arrived at from the opposite direction.

10 5emma_london, tammy_FL, Dr.LipidDallas and 7 others
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