Accepted answer
To be exact about it, dose arithmetic has three parts: concentration from vial content and diluent, volume from dose and concentration, and units from volume and syringe scale.
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.
Dead space by syringe type
| Configuration | Dead volume | Loss at 5 mg/mL | Over 20 draws |
|---|
| Fixed-needle insulin syringe | 3–5 µL | 15–25 µg | 0.3–0.5 mg |
| Low-dead-space, detachable | <2 µL | <10 µg | <0.2 mg |
| Standard luer-lock + 30G | 35–60 µL | 175–300 µg | 3.5–6 mg |
| Luer-lock + 21G drawing needle | 70–100 µL | 350–500 µg | 7–10 mg |
Worth being precise here: 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 Arrhenius relationship for drawing kinetics means that cold solution takes noticeably longer to draw than room-temperature solution.
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.
Do the arithmetic twice, ideally with someone else doing it independently.