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Does retatrutide adsorb measurably to the vial wall at 8 mg/mL?

Asked 4 Jun 2024Modified 22 months agoViewed 70k times
34

What I am working with: retatrutide · 8 mg/mL.

The units are where I keep going wrong, so please be explicit about them.

I have sanity-checked the order of magnitude and it seems right, which is not the same as being right.

Where is my error, and what is the correct working?

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RO
askedrae_oyelowo17k284 Jun 2024
8Same question here after a warm delivery, so I am following this. – deamidation_watch 4 months ago
Worth saying whether the vial has been opened, because that starts a different clock. – eoin_mcgarry 6 months ago
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5 Answers

Accepted answer first, then by votes
33

Accepted answer

8 mg/mL is 8000 µg/mL — roughly 800 times the concentration at which surface adsorption is measurable. Losses to glass and plastic matter in the low microgram-per-millilitre range, where a monolayer on the wall is a real fraction of what is in solution. At 8 mg/mL that same monolayer is a rounding error. If you see an apparent loss at this concentration, suspect the dilution step or the assay before you suspect the wall.

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

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.

Reported and extrapolated stability by condition

StateConditionUsable windowBasis
Lyophilised solid−20 °C, sealed, dry24–36 monthsSupplier guidance
Lyophilised solid2–8 °C, sealed12–24 monthsSupplier guidance
Lyophilised solid25 °C, sealed4–8 weeksExtrapolated (Arrhenius)
Lyophilised solid40 °C, sealed1–2 weeksExtrapolated
Solution, preserved2–8 °C28 daysUSP microbiological convention
Solution, preserved25 °C3–7 daysExtrapolated
Solution, unpreserved2–8 °C24 hoursUSP microbiological convention

Windows for the solid state are chemical; windows for solution are microbiological and usually shorter than the chemical limit.

Put another way, 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.

Metal-catalysed oxidation of methionine is documented across peptide and protein formulations and is why chelators appear in some formulations.

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

At dilute concentrations, suspect adsorption before you suspect chemistry.

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MO
answered · acceptedmarta_okonkwo190k25810 Sept 2024
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29

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

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.

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.

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

edited 26 Sept 2024 by harriet_lonsdale — expanded the table to cover the lower concentration

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HL
answeredharriet_lonsdale35k13830 Aug 2024
13

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

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

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.

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

The caveat is that none of these pathways can be seen by looking at a vial, and a clear solution can be substantially degraded.

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

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DV
answeredDr_Ilse_Vandenberg113k2482 Oct 2024
5I have kept vials both ways for a year and this matches what I saw. – rosa_mendieta 5 months ago
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12

Asparagine and glutamine are the deamidation risk, and methionine is the oxidation risk.

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.

Aggregation at air-liquid interfaces is established from surface-tension and particle-count studies and is the basis for anti-agitation handling guidance.

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

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P9
answeredplate_count_9k78k24816 Jul 2024
5Two lots stored differently, reassayed at a year — the difference was smaller than I expected. – forty_two_c 6 days ago
4Adding for future readers: the domestic leg after delivery is the part you control. – tandem_gradient 8 months ago
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-3

To be exact about it, this is answerable from the chemistry rather than from anecdote, which is unusual and welcome.

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.

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.

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DK
answeredDr_Tomas_Kral53k3821 Sept 2024
5Any published figure for how much a collapsed cake actually retains? – marta_okonkwo 8 months ago
6I would add a sentence about light, since tryptophan-containing sequences care. – v_ramaswamy 10 months ago
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Your answer

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