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ForumsCOA & Analytical TestingLC-MS/MS vs HPLC for peptide analysis — November 2025 Page 5

LC-MS/MS vs HPLC for peptide analysis — November 2025

Dr.SportsMedIN Mon, Jul 22, 2024 at 3:37 PM 51 replies 2,260 viewsPage 5 of 11
tom_AK
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Aug 9, 2024 at 10:38 AM#21
roxy_nash said:
My vendor review relevant to certificates of analysis: ordered 5 times from my current compounder.

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: 15.3 minutes (consistent with semaglutide reference standard)
Peak purity: 99.4%
Related substances: 0.4% (single impurity at 9.7 minutes — likely a synthesis intermediate)
Mass spec: m/z 4113.6 (matches expected molecular weight)

This is a clean result. The mass spec confirmation is particularly important — it proves the molecule is actually semaglutide and not a substitution.

Last edited: Aug 9, 2024 at 2:38 PM
42 15steve_okc, dave_SLC, FDA_TrackerJim and 39 others
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hank_denver
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Aug 16, 2024 at 6:37 AM#22

Adding the numbers, since they settle part of this. For anyone reading later: the numbers in this thread are worth checking against a primary source before you act on them, including mine. Half the figures circulating in this community trace back to a secondary summary that dropped a qualifier.

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

43 16raj_cambridge, ingrid_STO, pete_nash and 40 others
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nick_SD_fit
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Aug 23, 2024 at 2:32 AM#23

Following on from roxy_nash — and this may be the naive question:

Was that from a primary source or from a summary of one?

44 17EndoResFellow, PharmacoVig_BOS, SurmountFan_IN and 41 others
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Sigma-Aldrich — Research-Grade Standards

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emily_PDX
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Aug 29, 2024 at 10:23 PM#24
hank_denver said:
For anyone reading later: the numbers in this thread are worth checking against a primary source before you act on them, including mine.

This is where I part company with the consensus forming above. The objection nobody has made: this only works if the measurement is doing what we assume it is doing, and that assumption has not been tested here.

45 18Dr.KarenChen, Dr.NateNeph, PharmD_Rodriguez and 42 others
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BethLabQueen
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Sep 5, 2024 at 6:10 PM#25
hank_denver said:
For anyone reading later: the numbers in this thread are worth checking against a primary source before you act on them, including mine.

That is right for the common case. Whether it is right for the case being asked about depends on a detail that has not been given.

46 19wei_SG, cory_ATX, lori_vegas and 43 others
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