Feb 26, 2024 at 4:00 PM#1
How to read a Certificate of Analysis — complete guide
Posting this as an important update for the coa & analytical testing community. Please read carefully as this may affect your current plans.
I will keep this thread updated as new information becomes available. If you have additional information or corrections, please post below and I will incorporate confirmed updates into this OP.
Key points:
- This information is current as of the date posted
- Circumstances may change — always verify independently
- If you have questions, check if they have been answered in the replies before posting
Bookmarking this thread is recommended — it will be updated as the situation evolves.
— COA_Karl | COA & Analytical Testing
20 15matt_MKE, Dr.ReproEndo, lucas_SP_BR and 17 others
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Feb 26, 2024 at 5:08 PM#2
Walkthrough of a real COA (anonymized):
Here's what a Janoshik COA for semaglutide typically looks like. I'm using data from a recent community-submitted test:
```
CERTIFICATE OF ANALYSIS
Lab: Janoshik Analytical
Sample ID: JS-2026-01-4872
Date Received: January 14, 2026
Date Reported: January 21, 2026
Sample Description: White lyophilized powder, labeled "Semaglutide 5mg"
TEST METHOD RESULT SPECIFICATION
Identity ESI-MS Confirmed MW 4113.58 ± 2 Da
(Observed: 4113.4 Da)
Purity HPLC-UV 98.3% ≥ 95.0%
(220nm)
Content HPLC vs. std 4.87 mg 4.50 - 5.50 mg
(97.4% of label)
Residual Solvents GC-FID < LOQ Meets USP <467>
Water Content Karl Fischer 1.2% ≤ 5.0%
CONCLUSION: Sample meets all tested specifications.
```
How to interpret this:
- Identity confirmed at 4113.4 Da — that's within 0.2 Da of theoretical. This is semaglutide.
- 98.3% purity — excellent. The remaining 1.7% is likely minor synthesis impurities (truncated sequences, oxidized variants). Nothing unusual.
- 4.87mg content — 97.4% of the labeled 5mg. You're getting slightly less than advertised but well within acceptable range.
- Residual solvents below limit of quantitation — good. Means the lyophilization was thorough.
- 1.2% water content — normal for lyophilized peptide. Up to 5% is acceptable.
This is a solid COA. The product is what it claims to be, at the right purity and potency.
Last edited: Feb 26, 2024 at 6:08 PM
19 14TinaHashiRN, robert_kc, dan_philly and 16 others
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Feb 26, 2024 at 6:16 PM#3
This is incredibly helpful. A few clarifying questions:
1. When the purity says 98.3%, what exactly is the other 1.7%? Is it dangerous?
2. How do I know a COA is real and not fabricated? I've heard vendors can just make fake COAs in Photoshop.
3. What's the difference between a vendor-provided COA (from the manufacturer) and a third-party COA (from Janoshik, etc.)? Which should I trust more?
Last edited: Feb 27, 2024 at 12:16 AM
18 13mike_mod, SarahChen_PharmD, sarah.morrison and 15 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 StandardsFeb 26, 2024 at 7:24 PM#4
Let me tackle these:
1. The "other 1.7%":
In peptide synthesis, common impurities include:
- Deletion sequences — the peptide chain missing one or more amino acids
- Oxidized variants — methionine residues oxidized during synthesis or handling
- Truncated sequences — synthesis stopped before completion
- Deamidation products — asparagine converting to aspartate
- TFA (trifluoroacetic acid) — residual counterion from purification
- Acetic acid salt forms — if the peptide was acetate-exchanged
At 98.3% purity, the 1.7% impurities are at very low levels. For injectable peptides, the concern is whether any single impurity exceeds 0.5% and whether it's been identified. Reputable labs will flag any individual impurity above 0.5% on the chromatogram. At sub-1% levels each, these synthesis byproducts are generally considered safe for research purposes (and pharmaceutical-grade GLP-1 products have similar impurity profiles — they're just more thoroughly characterized).
2. Fake COAs:
This is a real problem. Ways to verify:
- Contact the lab directly. Janoshik has a verification page where you can enter the sample ID to confirm it's genuine. Use it.
- Check for consistency. A fake COA often has mismatched fonts, wrong test methods, or results that are suspiciously "perfect" (e.g., exactly 100.0% purity — real analytical data has decimal variation).
- Look for the chromatogram. A real COA usually includes the actual HPLC chromatogram image. Faking a realistic chromatogram is much harder than faking a table of numbers.
- Batch-specific data. A real COA has a unique sample ID and batch number. If a vendor uses the same COA for every batch, it's likely fake.
3. Vendor vs. third-party COA:
Always prioritize third-party testing. A vendor-provided COA is the manufacturer grading their own homework. It may be legitimate, but there's an inherent conflict of interest. Third-party labs like Janoshik, PeptideDetective, or Vanta have no financial stake in the product passing. That independence is what makes their results credible.
Best case scenario: the vendor provides their own COA AND a community member independently tests the same batch at a third-party lab, and the results agree. That's the gold standard of verification.
17 12ricardo_MIA, BrianDallas92, labquiet_amy and 14 others
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Feb 27, 2024 at 1:58 AM#5
Great post PeptideChemSF. I want to add some detail on reading HPLC chromatograms since the OP mentioned them:
What you're looking at: An HPLC chromatogram is a graph with time (minutes) on the X-axis and detector response (absorbance units or millivolts) on the Y-axis. Each "peak" represents a compound eluting from the column.
For semaglutide, expect:
- Main peak at a retention time typically between 12-18 minutes (depends on the method). This is the semaglutide.
- Small peaks before the main peak — these are more polar impurities (hydrophilic, elute faster)
- Small peaks after the main peak — these are less polar impurities or oxidized forms
- Solvent front at ~2-3 minutes — ignore this, it's just the injection solvent
What "98.3% purity by HPLC" means: The area under the main semaglutide peak represents 98.3% of the total area of ALL peaks. This is "area percent" or "area normalization" purity.
Important caveat: HPLC purity depends heavily on the method. A method that uses a short column, fast gradient, or UV detection at 280nm might miss impurities that would show up with a longer column, slower gradient, or detection at 220nm. This is why comparing purity numbers across different labs requires knowing the method details.
A "99.5% pure" result from one lab is not necessarily better than "97.8% pure" from another if they're using different methods. The lab using the more sensitive method might simply be detecting more impurities that the other lab missed.
Last edited: Feb 27, 2024 at 7:58 AM
16 11Dr.MetabolicMD, RetaRick_CA, JenPlateau and 13 others
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