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ForumsCOA & Analytical TestingEndotoxin testing methods — what worked for you? Page 3

Endotoxin testing methods — what worked for you?

amsterdam_pete Sat, Jan 3, 2026 at 11:50 AM 22 replies 1,163 viewsPage 3 of 5
SkepticalSean
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Jan 8, 2026 at 4:34 AM#11
jim_asheville said:
Inter-laboratory variation comes from real methodological differences, not sloppiness: which reference standard was used and how it was itself…

Genuinely useful, thank you. I had the facts and not the framework. Printing the relevant bit and taking it with me.

22 20DerekSJ_a1c, paige_pharma, emma_london and 19 others
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PedsEndoPhilly
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Jan 8, 2026 at 7:42 AM#12

Clinical perspective, offered as context rather than as advice.

Mass spectrometry confirmation for certificates of analysis: beyond HPLC purity, mass spec tells you the molecule's exact molecular weight. This confirms identity — not just purity.

Why this matters: HPLC can show you a "pure" compound, but if it's pure of the WRONG compound, purity is meaningless. Mass spec at m/z 4113.58 Da confirms it's actually semaglutide. My Janoshik report includes both HPLC and mass spec.

If your testing provider only offers HPLC, consider upgrading to include mass spec for at least your first batch from a new vendor.

Last edited: Jan 8, 2026 at 12:42 PM
23 21wendy_avl, jason_paloalto, Dr.LeslieOBGYN and 20 others
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Dr.RaviCardio
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Jan 8, 2026 at 10:50 AM#13
newstart_MO said:
Third-party testing services for certificates of analysis besides Janoshik: Janoshik: Gold standard for peptide community, ~$138, 10-14 day turnaround…

Vendor quality matters when discussing certificates of analysis. I've tested my last 5 batches through Janoshik and purity has ranged from 99.4% to 99.6%. 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.

24 22mark_tokyo, hans_munich, jason_sac26 and 21 others
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Sigma-Aldrich — Research-Grade Standards

Certified reference materials, analytical reagents, and research-grade standards for peptide verification. Trusted by laboratories worldwide.

Shop Reference Standards
dave_SLC
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Jan 8, 2026 at 1:58 PM#14
amsterdam_pete said:
They are not contradictory, they measure different things, and this is the single most common misreading of a certificate.

HPLC results interpretation for certificates of analysis: when you get your Janoshik report back, here's what the numbers mean:

  • Purity %: Should be >97%. Below 95% is concerning.
  • Retention time: Should match reference standard for the peptide
  • Related substances: Degradation products should be <2% total
  • Single impurity: No individual impurity should exceed 1%

My last batch: 99.7% purity, 0.1% total related substances. Clean chromatogram with single sharp peak at expected retention time. Exactly what you want to see.

25 23FranDenver, Dr.BariatricHTX, LindaRN_retired and 22 others
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Admin
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Jan 8, 2026 at 5:06 PM#15

Moderator note: leaving this open. It is being argued well and the disagreement is the useful part.

26 24PeptideSynthNJ, Dr.KarenChen, Dr.NateNeph and 23 others
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