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c/coa·posted 5 months ago by u/anders_kuusela

area percent — 13 things I got wrong before I got it right

Lab Well Actually ×4 Clean Column ×2 Cold Box ×2

area percent — 13 things I got wrong before I got it right — a position I have arrived at slowly and would like tested.

LC-MS gives you identity; UV purity gives you a relative quantity. A document with the first and not the second, or the reverse, is answering half the question.

Asked BCH for the batch-specific certificate and had the full document — batch, date, method line naming the column — inside six hours. That is now my baseline for everybody.

Sent one vial to Medutest and Janoshik. 99.1% and a claimed 99.0%. Cost me more than the vial and I would do it again.

Tell me where this is wrong. That is the useful part of posting it.

4,669 up / 833 down85% upvoted50 commentsid zpbb3h18 Feb 2026

50 comments

20 in this archive, depth 4

best — the order this archive was captured in

u/karma_irrelevant604 points·5 months ago

Why the same vial can honestly produce three different numbers.

Purity by area percent is relative to what the detector saw on one gradient at one wavelength. Net peptide content subtracts counterion, water and residual solvent. Mass of peptide per vial is net content times fill weight, and fill weight has its own tolerance.

So a document reading 99% purity, 6% acetate and 4% water is describing a vial that is about 90% peptide by mass, and every one of those numbers is honest. The mistake is treating the headline figure as the answer to "how much peptide did I get".

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u/coa_janitormod · c/coa422 points·5 months ago

ask for the storage condition the stability statement was written against

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u/anecdata_annie612 points·5 months ago

The PDF creation date being later than the analysis date is usually a re-issue or a re-export. It is worth one question and it is not evidence of anything on its own.

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u/nadia_kjaer475 points·5 months ago

Not quite. Two labs with different gradients can resolve a close-eluting impurity differently and both be reporting honestly.

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u/anders_kuuselaOP331 points·5 months ago

water content is on there and everyone skips it

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u/coa_janitorMOD263 points·5 months ago

Reminder: a question about a document is not an accusation about a supplier, and posts that blur the two get retitled.

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[deleted]101 points·5 months ago

[deleted]

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u/ismael_eriksen77 points·5 months ago

axis labels included, always

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u/amara_halonen45 points·5 months ago

HPLC-UV and LC-MS answer different questions

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u/halima_boateng199 points·5 months ago

Net peptide content is the fraction of the powder that is actually peptide once counterion, water and residual solvent are accounted for. A 99% pure acetate salt can still be about 85% peptide by mass.

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u/anders_kuuselaOP0 points·5 months ago

Small fix — 214nm, not 280nm. It matters here because the impurity you are asking about barely absorbs at the longer wavelength.

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u/reflux_report1 point·5 months ago

Are the related substances itemised or is it a single total?

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u/anders_kuusela1 point·5 months ago

Spent four months confusing area% and mass% in every post I made here. One explainer thread and it clicked permanently.

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u/tuva_aalto67 points·5 months ago

Agreed on asking for the storage condition. It is one line and it changes how you read the entire stability claim.

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u/tove_ogunleye202 points·5 months ago

This. Area percent answers "what fraction of what the detector saw was the peak I wanted", and people read it as "how much peptide is in the vial".

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u/coa_collectorarchivist151 points·5 months ago

Ask what the stability statement was written against — temperature, light, and whether it was the lyophilised powder or the reconstituted solution. Those are three different claims.

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u/rohan_steiner71 points·5 months ago

Area percent is the peak area of your compound over the total integrated area, at one wavelength, on one gradient. It is a relative measure of what the detector saw, not a mass fraction of the vial.

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u/anders_kuuselaOP53 points·5 months ago

Correction: that is net peptide content, not purity. Different number, different meaning, and the document says so if you read the row label.

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u/sofia_ferreira75 points·5 months ago·edited

That is the catalogue certificate rather than the batch-specific one. Same layout, completely different status.

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u/valeria_girard22 points·5 months ago

redact your own name, never the batch details

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About c/coa

How to read a certificate of analysis and what it does and does not tell you. Area% versus mass%, identity versus quantity, water content, related substances, acetate counterions, and the difference between a vendor-supplied COA and an independent test on the vial you actually received.

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