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Evidence-based GLP-1 & peptide discussion since 2023
ForumsCOA & Analytical TestingHPLC vs UPLC for peptide purity — 6 month update Page 3

HPLC vs UPLC for peptide purity — 6 month update

labquiet_amy Tue, Nov 12, 2024 at 6:30 AM 15 replies 1,855 viewsPage 3 of 3
Dr.NutriCornell
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Mar 2024
Ithaca, NY
Nov 14, 2024 at 5:29 AM#11
Dr.Martinez said:
My HPLC chromatogram analysis for certificates of analysis — I sent my batch to Janoshik and here's what the results showed: Main peak retention time:…

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: Nov 14, 2024 at 7:29 AM
23 21emily_PDX, Dr.SleepRoch, laura_annarbor and 20 others
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lori_vegas
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Aug 2024
Las Vegas, NV
Nov 15, 2024 at 8:39 PM#12
zoe_NC said:
Agreed, and mass spec answers a question purity does not: identity.

Saving this. It is the first explanation that did not require me to already understand it.

24 22bbq_ray_KC, oliver_london, tane_welly and 21 others
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MikeFit_NJ
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Apr 2024
New Jersey
Nov 17, 2024 at 11:48 AM#13
Dr.Martinez said:
My HPLC chromatogram analysis for certificates of analysis — I sent my batch to Janoshik and here's what the results showed: Main peak retention time:…

I tested my certificates of analysis batch through Janoshik — 97.3% purity. That's within acceptable range for compounded peptides. Turnaround was 12 business days.

The test costs ~$123 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.

25 23DeniseRN_TPA, SandraNC_45, Dr.EndoIndy and 22 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
NicoleRaleigh
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Aug 2024
Raleigh, NC
Nov 19, 2024 at 2:56 AM#14
labquiet_amy 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…

COA red flags for certificates of analysis — here's what to look for when reviewing a Certificate of Analysis:

  • No HPLC data: If there's no chromatography graph, it's not a real COA
  • Exact 100% purity: Suspicious — real testing always shows slight variation
  • No batch number: Can't trace back to specific production
  • No endotoxin testing: For injectables, this is essential
  • Template looks like it was made in Word: Professional labs have standardized formats

I have seen fake COAs that are literally photoshopped. Always verify independently if possible.

26 24VendorMark, COA_Karl, MikeFit_NJ and 23 others
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mike_mod
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Nov 20, 2024 at 6:03 PM#15

Moderator note: a post naming a supplier has been edited. Discuss suppliers in the review sections, not here.

Last edited: Nov 20, 2024 at 8:03 PM
27 0PurityPaulOR, MaxMetOK, MounjBrad and 24 others
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