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.
Two reports on the same product from two laboratories, four percentage points apart, and no method stated on either.
The four things to check on any report, in order: does it name your batch, who commissioned it, what method was used, and does it report content as well as purity. A report failing any of those is weak evidence regardless of the percentage on it.
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.
Dr.MetabolicMD said:Inter-laboratory variation comes from real methodological differences, not sloppiness: which reference standard was used and how it was itself…
Agreed, and mass spec answers a question purity does not: identity. High purity tells you one thing dominates the chromatogram, not that it is the molecule you ordered. A mass within about a dalton of expected is the identity check.
Sigma-Aldrich — Research-Grade Standards
Certified reference materials, analytical reagents, and research-grade standards for peptide verification. Trusted by laboratories worldwide.
Shop Reference Standardsricardo_MIA said:Two reports on the same product from two laboratories, four percentage points apart, and no method stated on either.
Same position here, arrived at the long way round. 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.
Clinical perspective, offered as context rather than as advice.
HPLC results interpretation for certificates of analysis: when you get your Janoshik report back, here's what the numbers mean:
- Purity %: Should be >97%. Below 95% is concerning.
- Retention time: Should match reference standard for the peptide
- Related substances: Degradation products should be <2% total
- Single impurity: No individual impurity should exceed 1%
My last batch: 99.7% purity, 0.1% total related substances. Clean chromatogram with single sharp peak at expected retention time. Exactly what you want to see.