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c/coa·posted 2 years ago by u/clara_restrepo

unpopular opinion: most of what gets said here about water content is guesswork

Discussion Receipts ×9 Slow Clap ×1 Cold Box ×2

unpopular opinion: most of what gets said here about water content is guesswork, and I am aware this is a minority view on this board.

UV at 214nm sees the peptide bond and picks up almost everything; at 280nm you are looking at aromatics only. Which wavelength the number came from is part of the number.

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.

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.

Please do not ask me what dose you should be on. I genuinely do not know and neither does anyone else here.

4,646 up / 1,113 down81% upvoted39 commentsid 4kab2y18 Dec 2023

39 comments

26 in this archive, depth 5

best — the order this archive was captured in

u/zaid_balogun204 points·2 years 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/receptor_bias_rick157 points·2 years ago

Area percent is the peak area of your compound over the total integrated area, at one wavelength, on one gradient.

zaid_balogun has the distinction right — one is a fraction of the detector signal, the other is a fraction of the mass.

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u/nadia_fonseca129 points·2 years ago

a certificate with no method line is decorative

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u/valeria_karlsen47 points·2 years ago

redact your own name, never the batch details

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[removed]31 points·2 years ago

[removed by moderator]

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u/nadia_fonseca186 points·2 years 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/cesar_ivaturi105 points·2 years ago

area% is not mass%, and the difference is the whole game

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u/anecdata_annie-37 points·2 years ago

net peptide content and purity are two different numbers on the same vial

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u/clara_restrepo73 points·2 years 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/cormac_pereira38 points·2 years ago

Careful — "the metadata looks odd" is a reason to ask a question, not a conclusion. Re-issues are ordinary.

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u/dead_space_doug40 points·2 years ago

Net peptide content is the fraction of the powder that is actually peptide once counterion, water and residual solvent are accounted for.

Disagree on this specific point. Two honest labs can resolve that impurity differently and report different numbers.

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u/nikhil_lindqvist0 points·2 years ago

Net peptide content is the fraction of the powder that is actually peptide once counterion, water and residual solvent are accounted for.

Adding the bit that always gets missed: the lot on the vial has to match the lot on the page, or none of this applies to your material.

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u/ghrp_history21 points·2 years ago

Right, and net peptide content is where the counterion and the water go. Both numbers can be honest and very different.

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u/india_generic_g16 points·2 years ago

Area percent or net peptide content? They answer different questions.

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u/certified_referenceQC0 points·2 years ago

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

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u/milos_vanhecke13 points·2 years ago

Right, and net peptide content is where the counterion and the water go.

This is the sentence to take away. The rest of the thread is footnotes to it.

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u/gallbladder_gary7 points·2 years 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/dario_vermeulen71 points·2 years ago

Pushing back a little: a missing field is usually presentation, not concealment. Ask for the full document before drawing conclusions.

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u/step_therapy_s22 points·2 years ago

Cosigning. The batch identifier is load-bearing, and a certificate that cannot be tied to the vial in your hand is not about your vial.

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u/camila_lindqvist14 points·2 years ago·edited

Cosigning.

Agreed, and the practical version is: ask for the itemised table. If it exists, you get it.

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u/clara_restrepoOP6 points·2 years ago

Does the lot on the vial match the lot on the page?

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u/janek_ferrari10 points·2 years ago

HPLC-UV and LC-MS answer different questions

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u/zeynep_zielinski6 points·2 years ago

axis labels included, always

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u/gallbladder_gary3 points·2 years ago

Agreed — the itemised related substances table is the difference between a document and a receipt.

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u/nabila_espinoza1 point·2 years ago

ask for the storage condition the stability statement was written against

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u/soren_nkemelu38 points·2 years 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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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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