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

[Lab] split one vial across Janoshik and Medutest — 98.7% and 98.3%

Labbranch of 10 comments

Short version of the title, which is already short: split one vial across Janoshik and Medutest — 98.7% and 98.3%. Longer version underneath. Since the title puts numbers in the shop window: 98.7% and 98.3%. Bought a second-hand instrument and learned more in six months of fixing it than in three years of reading…

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10 comments, started 1 year ago
u/elodie_grimaldi78 points·1 year 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/andres_restrepoOP30 points·1 year ago

What wavelength, and what was the gradient?

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u/andres_restrepoOP11 points·1 year ago

Correcting myself upthread: I said the gradient was linear and looking again it has a hold in the middle.

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u/ignacio_vanhecke7 points·1 year 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/andres_restrepo5 points·1 year ago

LC-MS for identity, UV for relative quantity

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u/rasmus_petrescu25 points·1 year ago

That figure cannot be quoted to two decimals off that baseline. The precision is not in the data.

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u/b12_baseline16 points·1 year ago

That figure cannot be quoted to two decimals off that baseline.

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

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u/marit_mensa32 points·1 year 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/hugo_bergstrom0 points·1 year ago

Same view — reporting the gradient is what makes a result checkable rather than merely stated.

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The analytical deep end: RP-HPLC methods, gradient design, integration decisions, LC-MS confirmation, amino-acid analysis, water content, and why two labs can report 98.1% and 96.4% on the same vial and both be right. Post the chromatogram, not the summary line.

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