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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.

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.

Where I think it is weakest: the follow-up is short relative to how long people actually take these drugs, so durability is an assumption here rather than a finding.

What I am after is whether a purity figure and a content figure can disagree without one of them being wrong. Tell me what I have not thought of.

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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PeptideChemSF
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Oct 19, 2024 at 3:06 AM#2
kevin_tulsa said:
The provenance questions are more useful than the numbers.

kevin_tulsa has the substance of this right. The condition it depends on is worth stating. 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.

27 22dave_SLC, FDA_TrackerJim, ricardo_MIA and 24 others
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DebRD_ATL
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Oct 19, 2024 at 3:26 AM#3
kevin_tulsa said:
The provenance questions are more useful than the numbers.

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.

26 21SleepDoc_PDX, RegAffairsDC, BiostatsBrad and 23 others
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Sigma-Aldrich — Research-Grade Standards

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Dr.SportsMedIN
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Oct 19, 2024 at 3:46 AM#4

This one has a reasonably settled answer, so here it is. 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.

25 20Dr.SurgeonPGH, rachel_ABQ, traveltech_sara and 22 others
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emily_PDX
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Oct 19, 2024 at 5:34 AM#5
PeptideChemSF said:
Inter-laboratory variation comes from real methodological differences, not sloppiness: which reference standard was used and how it was itself…

Can confirm. Same sequence, different timescale.

24 19PeptideSynthNJ, Dr.KarenChen, Dr.NateNeph and 21 others
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