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How do I get 12 units out of a vial reconstituted to 8 mg/mL without guessing?

Asked 13 Aug 2024Modified 20 months agoViewed 25k times
15

Concretely: 12 units · 8 mg/mL.

I have done this once and I suspect I got away with it rather than got it right.

For context: I keep records of every batch, every lot number and every result, so an answer that requires me to track something is fine.

What does a defensible version of this look like in practice?

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askedfelix_araya17k2813 Aug 2024
2Small correction: the units in the third paragraph should be micrograms, not milligrams. – e_dziedzic 5 months ago
Do you have a reference for the last claim? Not disputing it, just want to read it. – Dr_Jonas_Halvorsen 3 months ago
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5 Answers

Accepted answer first, then by votes
8

Accepted answer

Dose arithmetic has three parts: concentration from vial content and diluent, volume from dose and concentration, and units from volume and syringe scale.

Dead space quantified: a fixed-needle insulin syringe holds roughly 3 to 5 µL in the hub and needle after the plunger bottoms out. A luer-lock syringe with a detachable needle holds 35 to 100 µL depending on the hub design. At 5 mg/mL that is 15 to 25 µg lost per draw on the insulin syringe and 175 to 500 µg on the luer-lock — which over ten draws is the difference between losing a rounding error and losing half a milligram.

Concentration and unit conversion at a glance

VialDiluentConcentration0.25 mg0.5 mg1 mg2.5 mg
5 mg1 mL5 mg/mL5 u10 u20 u50 u
5 mg2 mL2.5 mg/mL10 u20 u40 u100 u
10 mg1 mL10 mg/mL2.5 u5 u10 u25 u
10 mg2 mL5 mg/mL5 u10 u20 u50 u
10 mg3 mL3.33 mg/mL7.5 u15 u30 u75 u

Units are U-100 insulin units, where 1 unit = 0.01 mL. Divide dose by concentration for millilitres, then multiply by 100.

The concentration you actually work with is label claim times content fraction divided by actual diluent volume, which is usually not the same as the nominal concentration because content is usually not 100 per cent and you rarely measure the diluent volume to 0.1 mL precision.

Published data on syringe dead space quantifies low-dead-space designs as retaining under 2 µL against 35 µL or more for conventional detachable-needle syringes.

One qualification: if your arithmetic and someone else's disagree by a factor of ten, one of you has made a unit error, and writing out the units at every step is the diagnostic.

If in doubt, use more diluent and accept the shorter usable window.

edited 2 Oct 2024 by n_takahashi — added the citation requested in comments

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NT
answered · acceptedn_takahashi36k3819 Sept 2024
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11

Concretely, the common error is getting the concentration right but then misreading the syringe scale, which is why checking the barrel marking rather than your memory matters.

Breaking it down further: if a 10 mg vial has 96.5 per cent content, you have 9.65 mg of peptide. Divide that by 2.00 mL and your concentration is 4.825 mg/mL, not 5.00 mg/mL, which is a 3.5 per cent systematic error in every dose calculation.

On filtration: a 0.22 µm syringe filter will remove particulates and organisms, and it will also adsorb a fraction of your peptide onto the membrane — with a low-binding PVDF or PES membrane the loss is typically a few per cent.

Worth noting: the concentration after reconstitution is not the same as the label claim, and most people do not account for the difference.

Write the arithmetic on the vial label. It costs nothing and removes the step where you reconstruct it from memory.

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answeredDr_Bram_Verhoeven85k24811 Oct 2024
3For what it is worth, my own result was within half a per cent of this. – j_wierzbicki 7 months ago
4Any reason this would differ for a longer peptide? – seamus_brady 8 months ago
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6

On the detail: the arithmetic only stops being confusing once you work it through once and see that it is straightforward.

Number of stopper piercings matters less than the gauge doing the piercing. A 30G or 31G needle through a butyl stopper leaves a track that reseals; a 21G or 18G drawing needle punches a core and can drop it into the solution.

The underlying point is that rotation of injection site is a tolerability measure, not a pharmacokinetic one, but if you are going to do it you might as well do it right.

The insulin-unit standard U-100 means 100 units per millilitre, so one unit is 0.01 mL — this is the conversion that trips up more people here than any other single piece of arithmetic.

I would flag the obvious failure mode: people get the concentration right, get the volume right, and then read the syringe against the wrong scale.

Do the arithmetic twice, ideally with someone else doing it independently.

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answeredcal_hennessy14k2723 Oct 2024
Worth adding that the method section is where the answer usually is. – leonid_marchuk 4 days ago
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2

It helps to be literal here: this is arithmetic, so let us do the arithmetic rather than argue about it.

Room temperature before drawing is worth the ten minutes. Cold solution is more viscous, draws slower, and condensation on a cold barrel makes it harder to read the meniscus.

If in doubt, use more diluent and accept the shorter usable window.

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TG
answeredtandem_gradient85k24830 Sept 2024
6Adding for future readers: the certificate should carry the lot number, not just a batch code. – kelvin_lam 5 months ago
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1

More usefully, the single most useful thing to do is write the arithmetic on the vial label, because you will reconstruct it from memory at an inconvenient moment if you do not.

The rounding error accumulates if you round too many times — rounding concentration to 5.0, rounding the dose volume to 0.1 mL, rounding the unit reading to 10 — and the safest approach is to work the full precision and round only the final answer.

The caveat is that this assumes the vial contains what the label says, and if the content assay has not been done, the arithmetic is precise about an unknown quantity.

Write the arithmetic on the vial label. It costs nothing and removes the step where you reconstruct it from memory.

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answerednine_point_nine45k13825 Nov 2024

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.

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