The Peptide CommonsEst. May 2024
Independent. We sell nothing and are affiliated with no manufacturer or pharmacy. Every moderation action is logged in public
Analytics · Impurities & related substances

About the Impurities & related substances category

Pinned Wiki
BN
bench_notesTL4 Moderator7 Mar 2026#1
Community wiki post. Any member at trust level 3 or above can edit this post; every edit is recorded. Last edited by mira.patel on 27 Jun 2026.
  • 12 Apr 2026 — bench_notes: Added the worked example and a unit label to the table header.
  • 27 Jun 2026 — mira.patel: Clarified the distinction that was causing repeat questions below.
Editors: bench_notes, mira.patel, hana.sato
Staff post. Actions described here are recorded in the public moderation log and may be challenged in Meta.

Deletion sequences, oxidation, deamidation, aggregates, residual solvents, and counter-ion content.

This post is a community wiki: any member at trust level 3 or above can edit it, and every edit is recorded with its author and a note. If the scope described here has drifted from what actually gets posted, edit it rather than starting a topic about it.

What belongs here

  • Questions specific to impurities & related substances, with the units and the context stated.
  • Reports and datasets, with the collection method stated.
  • Disagreement about the substance, conducted in public.

What belongs elsewhere

  • Anything urgent belongs in Safety, immediately, and not here.
  • Anything about how this site works belongs in Meta.
  • Requests for a maintained page belong in Wiki requests.

Before posting, it is worth checking the documentation commons and this category's tag pages. A great deal of what gets asked here has a maintained answer with a review date on it, which is more reliable than a two-year-old reply.

4 likes 5mo
HS
hana.satoTL4 Moderator30 Mar 2026#2
Staff post. Actions described here are recorded in the public moderation log and may be challenged in Meta.

Moderation notes for this category, so that nothing about how it is run is a surprise.

Topics get moved into and out of here regularly. A move is not a judgement — the category tree is not obvious from outside and putting a question in the wrong place is expected. When I move something I say so in the topic and the action appears in the public log with the guideline cited.

The two guidelines that come up most often in this category are R2, on supplying a source when asked, and R7, on category placement. Neither is used punitively.

8 likes 4mo
DS
d.szymanskiTL3Wiki editor16 Apr 2026#3

As a member rather than staff: the most useful thing I did when I started reading this category was to work through the tag pages rather than the topic list. The topic list shows you what is recent. The tags show you what is settled.

19 likes 3mo
SD
st.dialloTL2 Moderator1 May 2026#4

Suggestion for the wiki post above: it would be worth linking the two or three most-referenced topics in this category directly, since new members reliably ask for exactly those. Happy to make the edit myself if nobody objects, with a revision note.

0 likes 3mo
BW
br.wikstromTL2 Moderator15 May 2026#5
st.diallo, post #4: Suggestion for the wiki post above: it would be worth linking the two or three most-referenced topics in this category directly, since new members reliably ask for exactly those. Happy to make the edit myself if nobody objects, with a revision note. Go to post

Residual solvents: traces of solvents used in purification. These are usually tested by gas chromatography, not by HPLC. A specification for residual solvents should be stated separately from the purity.

0 likes in reply to #4 2mo
CE
crossover_entryTL3Regular29 May 2026#6
bench_notes, post #1: Deletion sequences, oxidation, deamidation, aggregates, residual solvents, and counter-ion content. This post is a community wiki: any member at trust level 3 or above can edit it, and every edit is recorded with its author and a note. If the scope described here has drifted from what actually gets posted, edit it rather than starting a… Go to post

Coming back to post #4, because the follow-up matters more than the original answer.

Acetate content: counter-ion content. Trifluoroacetate or acetate from the salt form of the peptide. Affects mass calculations and should be stated on a complete certificate.

4 likes in reply to #1 2mo
MM
m.marchettiTL2 Moderator10 Jun 2026#7

I disagree with the reply above, and I think the disagreement is substantive rather than terminological.

The distinction being drawn does not survive when you look at the published data for this specific question. I would be glad to be shown wrong on this, because the version I am arguing against is more convenient.

13 likes 2mo
GR
gradient_reviewTL2Member23 Jun 2026#8

Disulfide formation: if a peptide contains cysteine, it can form disulfide bonds with itself or with other molecules. Under oxidising conditions multiple species appear. Reducing conditions (like DTT) convert them back.

27 likes 1mo
NR
n.ramosTL2 Moderator4 Jul 2026 · edited#9

This follows post #6 rather than contradicting it.

Oxidation at methionine and tryptophan: adds 16 per oxygen. Usually elutes earlier. Oxidation is common in storage, especially if the solution is exposed to light or if antioxidants are not present.

7 likes 23d
MD
methods_draftTL2Member16 Jul 2026#10
gradient_review, post #8: Disulfide formation: if a peptide contains cysteine, it can form disulfide bonds with itself or with other molecules. Under oxidising conditions multiple species appear. Reducing conditions (like DTT) convert them back. Go to post

I read post #8 twice before replying, because I had assumed the opposite.

Deamidation at asparagine and glutamine: adds 1 approximately. Frequently appears as a close-eluting pair. It is a chemical modification that occurs during storage.

18 likes in reply to #8 12d
MI
m.ilungaTL2 Moderator27 Jul 2026#11

Incomplete deprotection: mass higher by the protecting group mass. Usually markedly later eluting. A synthesis artifact from incomplete removal of protecting groups.

2 likes 15h

Suggested topics

TopicParticipantsRepliesViewsActivity
Particulates in a reconstituted solution: identifying the likely source
Posting this under the heading it deserves: Particulates in a reconstituted solution: identifying the likely source Everything below is what sits behind that. Posting the method first, because I know what the…
SCNECCYMRM+15 19 13k 9mo
Trifluoroacetate content and its consequences
Trifluoroacetate content and its consequences — 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 front of me…
AIRDRVHFAV+123 134 31k 5mo
Racemisation and why it is invisible to a mass measurement — what changed since
Racemisation and why it is invisible to a mass measurement — what changed since Writing it up because I had to work it out twice and would rather nobody else did. Working through the identity arithmetic and I…
TVBVCCBIN+11 15 42k 16d
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
[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

Related topics — sharing the tags forced degradation, counter-ion content, water content

TopicParticipantsRepliesViewsActivity
Deamidation and the close-eluting pair it produces — the long version
Posting this under the heading it deserves: Deamidation and the close-eluting pair it produces — the long version Everything below is what sits behind that. I would like to understand what this number means…
KMMNOAHR 3 347 24d
Follow-up: Attributing a new peak: degradation product or excipient?
Attributing a new peak: degradation product or excipient? I have a specific reason for asking rather than idle curiosity, and the context is below. Posting the method first, because I know what the first…
KIABCSGV+15 19 671 17mo
Isotope patterns and why the monoisotopic peak is not the tallest — does this still hold?
Isotope patterns and why the monoisotopic peak is not the tallest — does this still hold? — that is the question, and I have not found it answered plainly anywhere I have looked. Working through the identity…
PODBTD 2 4.6k 7h
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
Deconvolution of a multiply charged envelope
Deconvolution of a multiply charged envelope — setting out what I have, and where I think it stops being reliable. Working through the identity arithmetic and I would like it checked. tirzepatide has a…
IDIAJV 2 63 10mo