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c/hplc·submitted 1 year ago by u/two_mil_or_one

[PSA] chromatogram is not what most of this community thinks it is

Discussionbranch of 8 comments

Something I noticed reading old threads that I have not seen said out loud. The advice on HPLC in this community changed substantially around the start of last year, and nobody went back and updated the older posts. So depending on which thread the search engine hands you, you get two contradictory answers with the…

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8 comments, started 1 year ago
u/ewan_tulloch14 points·1 year ago

Yeah, the chromatogram profile matters more than the headline number.

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u/anya_erdogan7 points·1 year ago

This is the correct method framing. Column and gradient are load-bearing fields.

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u/careful_gradient2 points·1 year ago·edited

Slight fix: the number was 99.0, not 98.9. Decimal point, but a fairly consequential one.

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u/reflux_report4 points·1 year ago

Detection wavelength? integration and that changes at different wavelengths.

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u/cormac_roos10 points·1 year ago

Inter-lab variance on this kind of assay is routinely 1–2 points. Different column, different gradient, different integration.

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u/rina_nascimento5 points·1 year ago

a round-robin on baseline integration would be genuinely useful

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u/ahmed_rasmussen3 points·1 year ago

Inter-lab variance on this kind of assay is routinely 1–2 points.

Disagree on that part. 97.4% and 96.2% on the same vial is normal.

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u/runa_cabrera3 points·1 year ago·edited

That is net peptide content, not purity. Different number, different meaning.

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