Accepted answer
More usefully, write the units at every step, because units errors are the failure mode that catches everyone eventually.
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.
Reading a lyophilised cake
| Appearance | Interpretation | Action |
|---|
| Intact opaque puck, proud of base | Cycle ran correctly | Proceed |
| Slumped to one side | Shipped before fully dry, or vibration | Usually usable; note it |
| Glassy translucent film | Collapse above glass transition | Test before use |
| Melt-back ring at stopper | Thermal excursion in transit | Test before use |
| No visible cake at all | Very low fill, or nothing there | Weigh it; query the supplier |
It helps to be literal here: 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.
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.
Worth noting: the concentration after reconstitution is not the same as the label claim, and most people do not account for the difference.
Do the arithmetic twice, ideally with someone else doing it independently.