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What happens to semaglutide after twelve weeks at room temperature in solution?

Asked 11 Apr 2024Modified 2.0 years agoViewed 32k times
11

What I am working with: semaglutide · twelve weeks · room temperature.

This is one of those things that everyone repeats and nobody derives.

This matters practically, not just academically, because it changes what I would do next.

What is the causal chain, and where does it stop being established?

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askedsunniva_dahl22k2711 Apr 2024
Same question here after a warm delivery, so I am following this. – laminar_bench 8 months ago
8Worth saying whether the vial has been opened, because that starts a different clock. – tabular_nums 6 months ago
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5 Answers

Accepted answer first, then by votes
90

Accepted answer

twelve weeks is 84 days, which at room temperature is on the order of 283 refrigerated days. Room temperature is not a number, so take the pharmacopoeial 20–25 °C and its 22.5 °C midpoint: 17.5 kelvin above the 5 °C middle of a 2–8 °C refrigerator. The ten-degree rule of thumb — degradation rate roughly doubling per 10 K — puts that at about 3.4 times the refrigerated rate. It is an order-of-magnitude statement about a rate, not a shelf life, and the top of the 20–25 °C band runs about 1.4 times faster than the bottom of it. In solution the routes that matter are hydrolysis of the backbone, deamidation at Asn, and physical association — the first two cost content, the third costs neither until it precipitates. Over 84 days at room temperature you should expect all three to have moved, and a purity figure to have noticed only some of them. Reconstituted material has no certificate; the one in the box describes the powder.

Start with the sequence, because which pathways are available depends on which residues are present.

Hydrolysis cleaves the backbone, most readily at aspartate-proline and aspartate-glycine sequences, and is acid-catalysed. In a dry solid it barely proceeds at all.

Aggregation is physical: peptides unfold at air-liquid interfaces and associate. Shaking maximises that interface, which is why swirling and shaking produce visibly different outcomes on the same vial.

Adsorption losses at low concentrations are quantified in formulation studies and are the reason carrier proteins are used in dilute preparations.

A mass spectrum names the pathway. Plus one, plus sixteen, minus eighteen.

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answered · acceptedten_mg_vial31k13831 Jul 2024
5This should be linked from the help pages. – tri_gly_ala 2 months ago
4Small correction: it is the number of cycles rather than the freezer temperature that does the damage. – mz_4113 10 months ago
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35

Answering this needs the physical state, since a dry powder is protected from most of these and a solution is protected from none.

Freeze-thaw cycling drives aggregation through concentration at the ice interface and pH shifts as buffer components crystallise out at different rates. Each cycle costs something.

A mass spectrum resolves most of this: minus eighteen is dehydration or succinimide, plus one is deamidation, plus sixteen is oxidation, and an unchanged mass with a shifted retention time is an isomer.

Nothing here is medical advice, and research-use compounds are not approved for human use.

Cold, dry, dark, still. Those four words cover most of the mitigation.

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EB
answeredelke_brunner17k2813 Apr 2024
25

Answer first: the degradation pathways worth knowing are hydrolysis, deamidation, oxidation, aggregation and adsorption, and each has a different trigger and a different mitigation.

Oxidation targets methionine, cysteine and tryptophan, adding sixteen daltons per oxygen. It is catalysed by trace metals and promoted by dissolved oxygen and by light.

Adsorption onto glass and plastic is significant at low concentrations — micrograms per millilitre — and negligible at milligrams per millilitre. It is the usual explanation for an apparent loss in a dilute preparation.

Apparent loss in a dilute preparation is usually adsorption rather than degradation and is worth ruling out first.

Swirl, never shake. Aggregation is a handling problem more than a time problem.

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GA
answeredgrainne_ahearn50k389 Jul 2024
Adding a vote because this deserves more of them. – esther_vandeVelde 5 months ago
I have kept vials both ways for a year and this matches what I saw. – thabo_maseko 3 months ago
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21

The honest answer is that most reported "degradation" is adsorption and dilution error rather than chemistry.

Light exposure matters for tryptophan-containing sequences and for anything with a chromophore. Amber vials and a closed box are free mitigations.

Deamidation via the succinimide intermediate is well characterised, with sequence-dependent rates highest for asparagine-glycine motifs.

Sequence decides which pathways are even available. Check the residues.

edited 23 Jun 2024 by forty_two_c — fixed an arithmetic slip in the third paragraph

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answeredforty_two_c66k5817 Jun 2024
20

The relevant point is that a mass shift of plus one dalton is deamidation and plus sixteen is oxidation, so degradation is often visible in a mass spectrum if anyone looks.

Deamidation converts asparagine or glutamine to the corresponding acid via a succinimide intermediate, adding one dalton. It is base-catalysed, accelerates above neutral pH and is the dominant aqueous pathway for many peptides.

At dilute concentrations, suspect adsorption before you suspect chemistry.

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MV
answeredmala_venkatesh22k3720 Jul 2024
6This should be in the site help pages rather than buried in an answer. – tabular_nums 4 months ago
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Your answer

Ask PeptideStack is a static archive. Posting is closed, but the norms are worth stating: answer the question that was asked, show your working, cite the trial or the certificate, and say plainly where the evidence runs out.

Not medical advice. Research-use-only compounds are not approved for human use.