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Evidence-based GLP-1 & peptide discussion since 2023
ForumsCOA & Analytical TestingHas anyone dealt with moisture content analysis for lyophilized peptides?

Has anyone dealt with moisture content analysis for lyophilized peptides?

newstart_MO Thu, May 30, 2024 at 8:42 AM 54 replies 2,788 viewsPage 1 of 11
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newstart_MO
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Springfield, MO
May 30, 2024 at 8:42 AM#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 why two laboratories report different purity for the same batch, and which number to believe.

I would rather have one careful answer than five confident ones.

21 16bri_stats, pete_manc_UK, anna.melb_AU and 18 others
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JessicaH_TX
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Dec 2023
Houston, TX
May 30, 2024 at 9:29 AM#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.

20 15sarah_TO, wendy_avl, jason_paloalto and 17 others
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carlos_SATX
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Oct 2024
San Antonio, TX
May 30, 2024 at 10:16 AM#3
JessicaH_TX said:
Inter-laboratory variation comes from real methodological differences, not sloppiness: which reference standard was used and how it was itself…

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.

Correct me if the detail matters more than I have assumed.

19 14Dr.BariatricHTX, LindaRN_retired, tommy_boulder and 16 others
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pam_stl
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Oct 2024
St. Louis, MO
May 30, 2024 at 11:03 AM#4
newstart_MO said:
Two reports on the same product from two laboratories, four percentage points apart, and no method stated on either.

Same position here, arrived at the long way round. 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.

Ask again with the specifics and you will get a better answer than this one.

18 13SleepDoc_PDX, RegAffairsDC, BiostatsBrad and 15 others
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pete_nash
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Aug 2024
Nashville, TN
May 30, 2024 at 3:28 PM#5

Clinical perspective, offered as context rather than as advice.

Vendor quality matters when discussing certificates of analysis. I've tested my last 6 batches through Janoshik and purity has ranged from 98.7% to 100.7%. Consistency is key.

For anyone new: always request the COA, and consider independent verification. The $300 for a Janoshik test is worth the peace of mind.

17 12sarah.morrison, NeuroNate, JessicaH_TX and 14 others
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