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c/hplc·posted 8 months ago by u/ferran_krastev

[Lab] MKM cagri — Medutest came back 97.7% against a claimed 97.0%

Lab Receipts ×7 Long Haul ×2 Sourced ×3

Right: MKM cagri — Medutest came back 97.7% against a claimed 97.0%. I paid for this one myself, nobody sent me anything, and the receipts are in the comments.

The relevant figures are 97.7% and 97.0%, and they come from the same log I have kept the whole time.

Retention time shifted 0.4 minutes and I assumed the worst. It was the column temperature.

Bought a second-hand instrument and learned more in six months of fixing it than in three years of reading traces.

If two or three other people have done the same thing we might actually learn something. Alone it is an anecdote.

5,855 up / 486 down92% upvoted54 commentsid zj1sfo2 Dec 2025
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54 comments

25 in this archive, depth 5

best — the order this archive was captured in

u/fatima_yildiz413 points·7 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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u/dead_space_doug-14 points·7 months ago

Reading a trace posted here, in the order I actually look at things.

Axes first — if the wavelength and the time axis are not labelled, I stop. Then the baseline: where has it been drawn, and does the drawing absorb anything. Then peak shape: fronting, tailing, a shoulder that never resolves. Then the blank, if one was run, for carryover.

Only after all of that do I look at the percentage, and by then I usually know how much weight it deserves. The number is the last thing on the page and the first thing everybody argues about, which is exactly backwards.

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[removed]1 point·7 months ago

[removed by moderator]

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u/coa_janitormod · c/coa1 point·7 months ago

system suitability before you believe any number on the run

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u/freya_baptista1 point·7 months ago

Did you run a blank between injections?

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u/marisol_frisk1 point·7 months ago

Did you run a blank between injections?

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

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u/ferran_krastevOP1 point·7 months ago

Chased a ghost peak for three weeks. It was the plumbing. It is always the plumbing.

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u/gustav_solberg1 point·7 months ago

Careful — you are treating retention time as identity. Two things can co-elute and the trace will not tell you.

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u/quiet_reader_991 point·7 months ago

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

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u/liv_okafor1 point·7 months ago

integration decisions move the number more than the sample does

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u/marisol_frisk1 point·7 months ago

What did system suitability look like on that sequence?

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u/ferran_krastevOP1 point·7 months ago·edited

Is that baseline drawn by the software or by hand?

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u/ahmed_rasmussen1 point·7 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/aksel_kjaer246 points·7 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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u/first_hundred280 points·7 months ago

The system suitability argument, since it comes up whenever somebody posts a number without one.

Before the sample result means anything, the instrument has to be shown to be fit that day: replicate injections with an acceptable RSD on area, a tailing factor inside limits, adequate plate count, and a resolution check between the pair you care about.

None of that is exotic and all of it is routine in a laboratory that reports for a living. Its absence does not mean a number is wrong; it means the number is unanchored, and unanchored numbers should not be quoted to two decimal places on this board.

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u/amara_haddad163 points·7 months ago

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

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u/peak_area_peteMOD119 points·7 months ago

Retitled: the original claimed a comparison the post does not actually make.

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u/anya_erdogan29 points·7 months ago

axis labels or the trace is decoration

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u/rina_bergstrom58 points·7 months ago

I would not blame the sample yet. Everything you have described is consistent with the column rather than the vial.

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u/baseline_drifteranalytical102 points·7 months ago·edited

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

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u/honest_syringe_2025113 points·7 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/ferran_krastevOP48 points·7 months ago

Homopeptide sent me the column, the gradient and the theoretical mass without being asked. That is a short list and they are on it.

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u/naomi_antonsen30 points·7 months ago

What wavelength, and what was the gradient?

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u/runa_cabrera53 points·7 months ago·edited

Resolution between two peaks depends on retention, selectivity and efficiency. A shallower gradient buys retention and usually resolution, at the cost of peak width and run time.

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