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c/hplc·posted 2 years ago by u/sofia_petrescu

[Lab] 16th independent test on ERP — 97.4% on a claimed 97.0%, and the trend is the interesting part

Lab Receipts ×9 Cold Box ×1 Long Haul ×1

Numbers in the title, numbers in the post. 16th independent test on ERP — 97.4% on a claimed 97.0%, and the trend is the interesting part.

Since the title puts numbers in the shop window: 97.4% and 97.0%.

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.

Sent the same vial to PeptideMeter and Medutest. 99.3% against a claimed 98.5%. The difference was the gradient, not the material.

Screenshot none of this. Read the whole thread, including the parts where I am told I am wrong.

4,609 up / 999 down82% upvoted50 commentsid ulplo12 May 2024
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50 comments

22 in this archive, depth 5

best — the order this archive was captured in

u/week_four_wall228 points·2 years 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/clara_danquah159 points·2 years ago

Area percent is the integrated area of your peak over the total integrated area at one wavelength on one gradient. Change any of those and the number changes without the sample changing.

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u/sofia_petrescuOP68 points·2 years ago·edited

214nm sees the peptide bond, 280nm sees the aromatics

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u/baseline_drifteranalytical104 points·2 years ago·edited

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

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u/sofia_petrescuOP61 points·2 years ago

What wavelength, and what was the gradient?

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u/sofia_petrescuOP32 points·2 years ago

What did system suitability look like on that sequence?

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u/first_hundred194 points·2 years ago

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

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

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u/chain_confirm_cpayments92 points·2 years ago

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

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/hamza_weiss74 points·2 years ago

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

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[deleted]29 points·2 years ago

[deleted]

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u/santiago_villalobos62 points·2 years ago

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

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u/nora_lundgren96 points·2 years ago

reproducibility beats resolution if you only get one of them

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u/hplc_hobbyistruns their own column52 points·2 years 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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u/samir_falk23 points·2 years ago·edited

Resolution between two peaks depends on retention, selectivity and efficiency.

Disagreeing with this bit: that spread is ordinary inter-lab variance, not a disagreement about the material.

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u/peak_area_peteMOD91 points·2 years ago

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

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u/insulin_syringe_ian39 points·2 years ago

Yes. Retention time is a hypothesis about identity. Mass is the answer.

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u/cormac_roos28 points·2 years ago

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

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u/valeria_cardoso18 points·2 years ago·edited

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

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u/crosspost_bot_no24 points·2 years 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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