The Peptide CommonsEst. May 2024
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Tags · Technique

HPLC

High-performance liquid chromatography, the workhorse purity method.

48 topics 16 solved Synonyms: uhplc, rp-hplc Group: Technique

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

TopicParticipantsRepliesViewsActivity
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…
NIBTDHFEF+87 101 27k 4h
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…
PNTDHFEFJR+112 120 7.6k 5h
[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…
HVKAMCDKR+69 76 920 6h
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…
MMSSNCEFGA+62 67 12k 9h
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…
JSPRMVMDEM+78 91 1k 10h
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…
GHAAJSLCCR+16 20 3.1k 11h
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…
FWNMGSTNH+16 20 11k 13h
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…
KBNKBNCPB 4 137 18h
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…
TNENDS 2 32k 1d
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…
MRDNLAPOBP+11 15 825 2d
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…
FDLTZYFWAA+98 114 37k 22d
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…
GHDBEVLSMP+4 8 7.2k 1mo
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…
NTKCTDOPA+120 127 41k 1mo
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…
NLPNV 2 15k 1mo
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…
MICRJNCBHK+43 47 36k 2mo
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…
ADSVCDFNAD+10 14 2.1k 2mo
[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…
BOECFA 2 59 2mo
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…
ESOFIASLC+37 41 7k 2mo
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…
NAKBMM 2 61k 3mo
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…
SCBBCN 2 59 3mo
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…
LCDSPOCLNK+54 59 14k 3mo
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…
VSCBAWSLEL+16 20 27k 3mo
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…
DNSGMB 2 22k 4mo
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…
DTCNFEGFZV+4 8 1.2k 4mo
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…
SABJBPJSMS+47 51 3.3k 5mo
[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…
CDASHKBSL+71 78 34k 5mo
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…
RVIDSSVKFP+61 69 2.9k 6mo
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…
DBKKKFFLA+35 40 22k 7mo
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…
LVNCOLNVMS+48 53 918 8mo
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…
MAMMC 2 15k 8mo