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ForumsCOA & Analytical TestingEndotoxin testing methods — what worked for you?

Endotoxin testing methods — what worked for you?

kevin_tulsa Sat, Oct 19, 2024 at 2:46 AM 37 replies 2,040 viewsPage 1 of 8
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kevin_tulsa
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Oct 19, 2024 at 2:46 AM#1

This gets cited here weekly, usually second-hand, so it is worth setting out what it does and does not establish.

They are not contradictory, they measure different things, and this is the single most common misreading of a certificate. Purity is a relative area measurement: the main peak as a percentage of total peak area in the chromatogram. Content is an absolute mass measurement against a reference standard. A vial can be 99% pure and contain 6.1mg — pure material, overfilled — or 99% pure and contain 3mg, which is pure material and a short fill. Purity tells you what fraction of what is there is the right molecule; content tells you how much of it there is. You need both, and a certificate with only purity is telling you half the story.

Where I think it is weakest: the completion rate deserves as much attention as the headline, because a large effect among those who finished is a different claim from a large effect among those enrolled.

So the question, as narrowly as I can put it: why two laboratories report different purity for the same batch, and which number to believe. I have searched first, so if this is covered somewhere point me at it and I will read it.

Note on sourcing:
Figures above are from the primary publication rather than the press summary. If a number here disagrees with one you have, post yours and we will work out which of us is reading a secondary source.
28 23fiona_glasgow, Dr.RheumBOS, greg_boulder and 25 others
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VendorMark
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Oct 19, 2024 at 3:06 AM#2
kevin_tulsa said:
They are not contradictory, they measure different things, and this is the single most common misreading of a certificate.

That is correct as far as it goes, and here is where it stops going. The provenance questions are more useful than the numbers. Does the report name the batch on your vial. Is the date consistent with when that batch existed. Is it addressed to the vendor or to the person who paid for the test. Was the sample taken from general stock or supplied by the vendor. A perfect certificate for a different batch is worth nothing, and vendor-supplied samples are the most common way a genuine report ends up misleading.

27 22KristenIndy, MarkLI_maint, Dr.PeteFamMed and 24 others
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PharmacoVig_BOS
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Oct 19, 2024 at 3:26 AM#3
kevin_tulsa said:
They are not contradictory, they measure different things, and this is the single most common misreading of a certificate.

I am more sceptical of certificates as a category than this thread is. The document tells you about a sample somebody chose to send, at a moment in time, and nothing about the vial in your hand. Independent purchase-and-test is a different quality of evidence and everything else is inference.

Last edited: Oct 19, 2024 at 6:26 AM
26 21DataDave, Dr.GutHealth, amsterdam_pete and 23 others
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roxy_nash
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Oct 19, 2024 at 3:46 AM#4

Answering the narrow version, because the broad one does not have a single answer. Inter-laboratory variation comes from real methodological differences, not sloppiness: which reference standard was used and how it was itself calibrated, the gradient and column, the detection wavelength, and where the integration baseline was drawn. A percentage point or two between competent labs is expected. What is not expected is a large gap, and the usual explanation there is that one report is not of the batch in your hand.

Last edited: Oct 19, 2024 at 9:46 AM
25 20Dr.NephBHM_UK, kim_atl_prep, sarah_TO and 22 others
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GraceAZ_72
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Oct 19, 2024 at 5:34 AM#5
VendorMark said:
The provenance questions are more useful than the numbers.

This is my experience too, for whatever a second data point is worth. I had assumed I was the exception until I read this.

Last edited: Oct 19, 2024 at 9:34 AM
24 19RetaRick_CA, JenPlateau, SallyK_inj and 21 others
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