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ForumsCOA & Analytical TestingInter-lab variability in peptide testing — July 2024

Inter-lab variability in peptide testing — July 2024

PedsEndoPhilly Fri, Jan 31, 2025 at 9:32 PM 52 replies 2,419 viewsPage 1 of 11
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PedsEndoPhilly
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Jan 31, 2025 at 9:32 PM#1

Two reports on the same product from two laboratories, four percentage points apart, and no method stated on either.

The question I want answered is whether a purity figure and a content figure can disagree without one of them being wrong.

Not looking for reassurance. Looking for the part I have got wrong.

37 7sarah_TO, wendy_avl, jason_paloalto and 34 others
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COA_Karl
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Jan 31, 2025 at 9:54 PM#2

This one has a reasonably settled answer, so here it is. 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.

I would rather be corrected than agreed with, if it comes to it.

Last edited: Jan 31, 2025 at 10:54 PM
36 6jim_asheville, matt_MKE, Dr.ReproEndo and 33 others
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tammy_FL
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Tampa, FL
Jan 31, 2025 at 10:16 PM#3
COA_Karl said:
Inter-laboratory variation comes from real methodological differences, not sloppiness: which reference standard was used and how it was itself…

That is correct as far as it goes, and here is where it stops going. One addition: an unstated method makes a report close to uninterpretable. "99% pure" without the technique, the standard and the conditions is a claim, not a measurement.

Happy to go further on any of that.

Last edited: Feb 1, 2025 at 4:16 AM
35 5kevin_tulsa, Dr.PainCLE, mike_mealprep and 32 others
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nick_SD_fit
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Sep 2024
San Diego, CA
Jan 31, 2025 at 10:38 PM#4
PedsEndoPhilly said:
Two reports on the same product from two laboratories, four percentage points apart, and no method stated on either.

This matches mine closely enough to be worth saying so. 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.

34 4PharmacoVig_BOS, SurmountFan_IN, PeptideChemSF and 31 others
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Dr.EndoIndy
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Indianapolis, IN
Feb 1, 2025 at 12:38 AM#5

From the other side of the consultation, briefly.

I tested my certificates of analysis batch through Janoshik — 97.3% purity. That's within acceptable range for compounded peptides. Turnaround was 14 business days.

The test costs ~$138 but gives you actual HPLC chromatography data plus mass spec confirmation. For the peace of mind alone, it's worth every penny. I test every new batch from a new vendor.

33 3SandraNC_45, Dr.EndoIndy, tom_AK and 30 others
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