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c/hplc·submitted 4 months ago by u/yara_bakken

[Discussion] we are measuring method development at the wrong time and calling it noise

Discussionbranch of 26 comments

we are measuring method development at the wrong time and calling it noise. I have gone back and forth on this for months. Why two honest labs report different numbers on the same vial. Start with the gradient. A shallower slope holds compounds on the column longer and usually separates close-eluting species better.…

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26 comments, started 4 months ago
u/peak_area_peteMOD393 points·4 months ago

Trace posted without axis labels — asked for a relabelled version rather than removing it.

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u/step_count_stan123 points·4 months ago

system suitability before you believe any number on the run

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u/saskia_bakker82 points·4 months ago

That is area percent, not mass percent. The trace cannot give you the second one.

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u/yara_bakkenOP45 points·4 months ago

Is that baseline drawn by the software or by hand?

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u/erez_yilmaz53 points·4 months ago·edited

Correction: that is a tailing factor, not a plate count. Different diagnostics for different problems.

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u/mariam_cabrera43 points·4 months ago

a blank injection between samples costs four minutes and settles most arguments

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u/naomi_antonsen69 points·4 months ago

system suitability before you believe any number on the run

Adding one practical thing — run the blank. It answers this before anyone has to argue about it.

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u/hplc_hobbyistruns their own column55 points·4 months ago

reproducibility beats resolution if you only get one of them

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u/week_four_wall274 points·4 months ago

Started reporting my own integrations with the baseline choice stated. Arguments in my threads dropped by about half.

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u/samir_falk98 points·4 months ago

This. A shoulder that does not baseline-resolve is a question, not a quantity.

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u/medutest_mel47 points·4 months ago

Cosigning the wavelength point. Half the disagreements in this board are two people looking at different detectors.

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u/anya_erdogan137 points·4 months ago

Gave the same trace to two people I trust and got two integrations about a point apart. That was the most educational afternoon I have had here.

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[removed]101 points·4 months ago

[removed by moderator]

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u/hedda_ekstrom394 points·4 months ago

axis labels or the trace is decoration

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u/andres_restrepo184 points·4 months ago

axis labels or the trace is decoration

This is the whole methodological point of the board in one line.

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u/santiago_villalobos275 points·4 months ago

A shoulder that does not baseline-resolve cannot be quantified honestly. You can report it as an unresolved shoulder, which is useful information, or you can develop the method until it resolves.

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u/honest_syringe_pls105 points·4 months ago

Right. And a blank between injections settles the carryover argument before it starts.

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u/hamza_weiss209 points·4 months ago

What did system suitability look like on that sequence?

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u/two_mil_or_one56 points·4 months ago

Carryover from a previous high-concentration injection looks exactly like a small impurity. The blank injection is four minutes and it removes the ambiguity entirely.

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u/fatima_yildiz33 points·4 months ago

Do you have the mass, or only the UV trace?

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u/adaeze_batista23 points·4 months ago·edited

Small fix — 214nm, not 210. It matters for the comparison you are making with the other run.

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u/yara_bakkenOP9 points·4 months ago

Did you run a blank between injections?

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