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

Dr.PulmRoch said:

Reconstituted peptide stability data for certificates of analysis: I had Janoshik test my reconstituted vial at day 1, day 14, and day 28 stored in…

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: 15.2 minutes (consistent with semaglutide reference standard)
Peak purity: 99.4%
Related substances: 0.8% (single impurity at 9.5 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.

jennifer_SEA said:

One addition: an unstated method makes a report close to uninterpretable.

Thank you — that is the clearest version of this I have read, and I have read a lot of them. I will report back once I have actually tried it.

Dr.PulmRoch said:

Reconstituted peptide stability data for certificates of analysis: I had Janoshik test my reconstituted vial at day 1, day 14, and day 28 stored in…

Community-verified vendors for certificates of analysis: I maintain a spreadsheet of Janoshik-verified test results submitted by community members. Across 50 submitted tests:

• 82% tested at ≥98% purity
• 12% tested at 95-98% purity
• 6% tested at <95% purity (concerning)

The vendors with consistently high purity results are listed in the Vendor Directory. We update monthly based on new community submissions.

Dr.SportsMedIN said:

Two reports on the same product from two laboratories, four percentage points apart, and no method stated on either.

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: 99.5% purity, 0.3% total related substances. Clean chromatogram with single sharp peak at expected retention time. Exactly what you want to see.

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