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c/hplc·submitted 10 months ago by u/ewan_zielinski

[Discussion] the chromatogram advice in here is 2 years out of date

Discussionbranch of 8 comments

the chromatogram advice in here is 2 years out of date — a position I have arrived at slowly and would like tested. Numbers, in the order they matter: 2 years. Resolution between two peaks depends on retention, selectivity and efficiency. A shallower gradient buys retention and usually resolution, at the cost of peak…

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8 comments, started 10 months ago
u/elodie_grimaldi231 points·10 months ago

Mass spectrometry answers identity. UV purity answers relative quantity under the run conditions. A document with one and not the other is answering half the question, and the half it answers should be stated.

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u/nabila_espinoza68 points·10 months ago

Agreed, and it is why MKM printing the column and gradient on the certificate is genuinely useful rather than decorative.

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u/fatima_yildiz22 points·10 months ago

two labs, two gradients, two honest answers

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u/dead_space_doug43 points·10 months ago

Agreed, and it is why MKM printing the column and gradient on the certificate is genuinely useful rather than decorative.

nabila_espinoza is right — the integration choice is a decision and it should be stated alongside the result.

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u/ewan_zielinskiOP32 points·10 months ago

you cannot report to two decimals off that baseline

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u/downvote_magnet58 points·10 months ago

baseline choice is a decision, not a measurement

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u/endotoxin_elliemicro27 points·10 months ago·edited

baseline choice is a decision, not a measurement

Agreed. And the corollary is that the method line on a certificate is not decoration, it is the number’s provenance.

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u/rasmus_kimani45 points·10 months ago

UV response is not uniform across species. At 214nm you are looking at the peptide bond, which is why it is the working wavelength for this class; at 280nm you only see tryptophan, tyrosine and phenylalanine.

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