Tags · Technique
HPLC
High-performance liquid chromatography, the workhorse purity method.
Related in this group: LC-MS · high-resolution MS · MALDI-TOF · UV & DAD detection · peak integration · system suitability · gradient method · water content · amino acid analysis · ELISA · endotoxin testing · sterility testing
48 topics · page 1 of 2
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What a reversed-phase purity number actually is
The question in the title: What a reversed-phase purity number actually is I will give what I have already checked below so nobody repeats it. I would like to understand what this number means before I repeat…
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+87 | 101 | 27k | 4h |
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Named, unnamed and unspecified impurities as regulatory categories — the long version
Named, unnamed and unspecified impurities as regulatory categories — the long version — setting out what I have, and where I think it stops being reliable. A documentation question rather than an analytical…
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+112 | 120 | 7.6k | 5h |
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[2026 update] Detection wavelength and why 214 nm and 280 nm disagree
Detection wavelength and why 214 nm and 280 nm disagree — setting out what I have, and where I think it stops being reliable. Posting the method first, because I know what the first three replies will…
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+69 | 76 | 920 | 6h |
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Follow-up: Comparing two chromatograms from different laboratories, properly
Comparing two chromatograms from different laboratories, properly — setting out what I have, and where I think it stops being reliable. I would like to understand what this number means before I repeat it…
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+62 | 67 | 12k | 9h |
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Endotoxin and why it is a separate question from purity — what changed since
On the subject in the title: Endotoxin and why it is a separate question from purity — what changed since Working notes rather than a conclusion. A documentation question rather than an analytical one. I have…
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+78 | 91 | 1k | 10h |
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Why 99.2% and 97.8% on the same lot can both be correct
Why 99.2% and 97.8% on the same lot can both be correct I have a specific reason for asking rather than idle curiosity, and the context is below. I would like to understand what this number means before I…
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+16 | 20 | 3.1k | 11h |
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Shoulders, tangent skims, and defensible integration
Posting this under the heading it deserves: Shoulders, tangent skims, and defensible integration Everything below is what sits behind that. Posting the method first, because I know what the first three…
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+16 | 20 | 11k | 13h |
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About the HPLC & UHPLC category
Reversed-phase separations, gradients, columns, integration, and reading a chromatogram properly. This post is a community wiki: any member at trust level 3 or above can edit it, and every edit is recorded…
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4 | 137 | 18h | |
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Gradient steepness and apparent purity: a worked comparison — the long version
Gradient steepness and apparent purity: a worked comparison — the long version — setting out what I have, and where I think it stops being reliable. A documentation question rather than an analytical one. I…
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2 | 32k | 1d | |
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Impurity thresholds: where the common numbers come from
Impurity thresholds: where the common numbers come from — setting out what I have, and where I think it stops being reliable. A documentation question rather than an analytical one. I have a certificate in…
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+11 | 15 | 825 | 2d |
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Carryover and the ghost peak from last week's standard — what changed since
Carryover and the ghost peak from last week's standard — what changed since Writing it up because I had to work it out twice and would rather nobody else did. Posting the method first, because I know what the…
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+98 | 114 | 37k | 22d |
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Second pass at: Where impurities in solid-phase peptide synthesis come from
Second pass at: Where impurities in solid-phase peptide synthesis come from Writing it up because I had to work it out twice and would rather nobody else did. A documentation question rather than an…
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+4 | 8 | 7.2k | 1mo |
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Racemisation and why it is invisible to a mass measurement
Posting this under the heading it deserves: Racemisation and why it is invisible to a mass measurement Everything below is what sits behind that. Working through the identity arithmetic and I would like it…
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+120 | 127 | 41k | 1mo |
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Resolution and tailing factor: real acceptance criteria
Resolution and tailing factor: real acceptance criteria Writing it up because I had to work it out twice and would rather nobody else did. I would like to understand what this number means before I repeat it…
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2 | 15k | 1mo | |
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Injection volume, overload, and peak distortion
Posting this under the heading it deserves: Injection volume, overload, and peak distortion Everything below is what sits behind that. A documentation question rather than an analytical one. I have a…
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+43 | 47 | 36k | 2mo |
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Area percent versus weight percent: the confusion that causes most arguments
Posting this under the heading it deserves: Area percent versus weight percent: the confusion that causes most arguments Everything below is what sits behind that. Posting the method first, because I know…
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+10 | 14 | 2.1k | 2mo |
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[2026 update] Reading a chromatogram someone posted without axes
Posting this under the heading it deserves: Reading a chromatogram someone posted without axes Everything below is what sits behind that. A documentation question rather than an analytical one. I have a…
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2 | 59 | 2mo | |
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Residual solvents and scavengers: what is looked for — what changed since
Residual solvents and scavengers: what is looked for — what changed since — setting out what I have, and where I think it stops being reliable. Posting the method first, because I know what the first three…
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+37 | 41 | 7k | 2mo |
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Impurity thresholds: where the common numbers come from — what changed since
Posting this under the heading it deserves: Impurity thresholds: where the common numbers come from — what changed since Everything below is what sits behind that. A documentation question rather than an…
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2 | 61k | 3mo | |
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Deletion sequences: expected mass shifts by residue
Deletion sequences: expected mass shifts by residue — setting out what I have, and where I think it stops being reliable. Working through the identity arithmetic and I would like it checked. semaglutide has a…
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2 | 59 | 3mo | |
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Where impurities in solid-phase peptide synthesis come from
Where impurities in solid-phase peptide synthesis come from Writing it up because I had to work it out twice and would rather nobody else did. A documentation question rather than an analytical one. I have a…
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+54 | 59 | 14k | 3mo |
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Shoulders, tangent skims, and defensible integration — the long version
Shoulders, tangent skims, and defensible integration — the long version Writing it up because I had to work it out twice and would rather nobody else did. Posting the method first, because I know what the…
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+16 | 20 | 27k | 3mo |
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Endotoxin and why it is a separate question from purity
Endotoxin and why it is a separate question from purity Writing it up because I had to work it out twice and would rather nobody else did. I would like to understand what this number means before I repeat it…
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2 | 22k | 4mo | |
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Injection volume, overload, and peak distortion — one year on
Injection volume, overload, and peak distortion — one year on Writing it up because I had to work it out twice and would rather nobody else did. A documentation question rather than an analytical one. I have…
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+4 | 8 | 1.2k | 4mo |
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Second pass at: Where impurities in solid-phase peptide synthesis come from
Posting this under the heading it deserves: Second pass at: Where impurities in solid-phase peptide synthesis come from Everything below is what sits behind that. A documentation question rather than an…
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+47 | 51 | 3.3k | 5mo |
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[2026 update] A rising related-substance total over six months: degradation or method drift?
The question in the title: A rising related-substance total over six months: degradation or method drift? I will give what I have already checked below so nobody repeats it. Posting the method first, because…
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+71 | 78 | 34k | 5mo |
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Aggregates and why a purity assay may miss them entirely
Aggregates and why a purity assay may miss them entirely Writing it up because I had to work it out twice and would rather nobody else did. A documentation question rather than an analytical one. I have a…
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+61 | 69 | 2.9k | 6mo |
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Comparing two chromatograms from different laboratories, properly
Posting this under the heading it deserves: Comparing two chromatograms from different laboratories, properly Everything below is what sits behind that. I would like to understand what this number means…
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+35 | 40 | 22k | 7mo |
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Revisiting: Aggregates and why a purity assay may miss them entirely
Posting this under the heading it deserves: Revisiting: Aggregates and why a purity assay may miss them entirely Everything below is what sits behind that. I would like to understand what this number means…
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+48 | 53 | 918 | 8mo |
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What a reversed-phase purity number actually is — the long version
What a reversed-phase purity number actually is — the long version I have a specific reason for asking rather than idle curiosity, and the context is below. I would like to understand what this number means…
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2 | 15k | 8mo |