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ForumsCOA & Analytical TestingUSP reference standards for peptide verification — anyone have experience?

USP reference standards for peptide verification — anyone have experience?

lisa_labSD Tue, Sep 24, 2024 at 11:05 AM 13 replies 1,991 viewsPage 1 of 3
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lisa_labSD
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Sep 24, 2024 at 11:05 AM#1

I have a certificate reporting 99.2% purity and 6.1mg content on a vial sold as 5mg, and I cannot work out whether those two numbers are consistent or contradictory.

What I am trying to establish is why two laboratories report different purity for the same batch, and which number to believe.

Numbers rather than impressions, if you have them.

28 23stefan_berlin, Dr.EM_Chicago, pete_RVA and 25 others
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HPLC_Greg
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Research Triangle, NC
Sep 24, 2024 at 11:20 AM#2

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

27 22JenPlateau, SallyK_inj, CryptoCarl and 24 others
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tom_AK
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Sep 24, 2024 at 11:35 AM#3
HPLC_Greg said:
The provenance questions are more useful than the numbers.

Agreed, and mass spec answers a question purity does not: identity. High purity tells you one thing dominates the chromatogram, not that it is the molecule you ordered. A mass within about a dalton of expected is the identity check.

26 21tyler_CSCS, VanRx_Mike, steve_okc and 23 others
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tommy_boulder
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Boulder, CO
Sep 24, 2024 at 11:50 AM#4
lisa_labSD said:
I have a certificate reporting 99.2% purity and 6.1mg content on a vial sold as 5mg, and I cannot work out whether those two numbers are consistent or…

Can confirm the pattern lisa_labSD describes. 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.

Last edited: Sep 24, 2024 at 12:50 PM
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sophie_paris
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Sep 24, 2024 at 1:12 PM#5

Adding the clinical framing, because it changes how the question reads.

I tested my certificates of analysis batch through Janoshik — 99.8% purity. That's within acceptable range for compounded peptides. Turnaround was 12 business days.

The test costs ~$143 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.

24 19BethLabQueen, ChrisMacros, KetoKyle and 21 others
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