Large Guide: Dispense
Your Setup
Youâll be working in front of your laminar flow hood for this whole procedure. You are not working in a still air box, as youâre processing far too many vials to not have a fully sterile environment.
You have stoppers in a sterile container that you can open in front of the laminar flow so they will stay sterile.
You have multiple wrapped sterile tweezers that you will use to pick up the stoppers. If you accidentally drop one or touch it to something non-sterile you can swap it out for one of your sterile, unused ones.
All of your vials are sterile and still have foil hats on them. They are staged in a way that you have ready access to them as you need them.
I prefer to dispense with a 20mL syringe with a 18g needle attached. You may have bigger hands than me and like a larger syringe. I find that the marks on the side of the 20mL syringe are easy to read.
- My 100mL vials are ready to go.
- Spare 20mL syringes, ready.
- Spare 18g needles, ready.
- Alcohol swabs, ready.
- 70% IPA spray, ready.
- Paper towels, ready.
- Vials caps, ready.
- Vial crimper, ready.
My laminar flow hood has been running, in my tent with a positive pressure environment, for at least an hour in order to get the ambient air in the tent as clean as possible.
I have wiped my whole work area down with my anti-bacterial, anti-viral cleaner.
If my laminar flow hood requires it, my work area in front of the flow hood is slightly elevated so that all my work is happening in front of the flow.
At no point in this process will I have to get up and leave. I wonât need to dig through any boxes. Everything I need is staged, and anything I wonât need is moved well out of the way.
My task light is on so I can see clearly.
I have a helper, Person 2, sitting next to me who understands how this is all about to go down. Person 1 will do the bulk of the aseptic work.
You are wearing full PPE.
Working in Front of Laminar Flow
This is a discussion that exists here and here. Please donât skip. Perfect aseptic technique is an essential part of how our vials end up sterile. Make sure your helper understands this as well.
If you work on a riser in front of the flow hood, make sure itâs not positioned so that if something spills it can fall onto the filter.
Dispensing
Iâm going to outline a recommended way to work. You may find a workflow that you prefer more.
Iâm assuming youâre using 3mL vials and 20mL syringes, adjust as needed.
Person 2 opens the 20mL syringe / 18g needle packaging in the sterile field. They attach the two. They draw up 18mL of air from the sterile field. They use an alcohol swab to clean the stopper of a 100mL vial. The inject 18mL of sterile air, and draw up 18mL of sterile preparation, without large bubbles. They cap the needle. They do this again with another syringe.
We use 18mL as it is divisible by 3, and weâre using 3mL vials.
Person 1 takes the syringe and in the sterile field removes the cap. They hold the syringe in hand while removing the hat from a sterile vial. Trying not to touch the needle to the side of the vial (difficult for a 3mL vial) they inject 3mL into the vial while still holding it in their hands.
They set the open vial down near the filter of the flow hood to minimize risk of anything accidentally being set in front of it.
18mL -> 15mL -> 12mL -> 9mL -> 6mL -> 3mL -> 0mL
Person 1 does one vial at a time, lining them up across the sterile field. They are not placed in front of each other, but next to each other. After theyâre finished they can set the syringe down, and Person 2 can fill it back up.
Person 1 takes the tweezers and gently places a stopper on each vial thatâs been lined up.
Either Person 1 or Person 2 can use their clean, gloved finger to press the stopper into place. Person 2 can place the caps on top of the stoppers and crimp them.
Person 1 continues in the same fashion, filling vials and placing stoppers.
Person 2 continues filling syringes and crimping vials.
We start with brand new needles and syringes as often as possible with the economics of your setup. Changing them out every time you start on a new 100mL vial would be a decent benchmark, meaning each would get used for dispensing about 3 time.
Crimping does not have to take place in the sterile field if you have limited room. Once the stopper is pressed down into place the vial is technically sealed. Sometimes pressure in the vial will cause the stopper to pop out of place. If this happens while not in the sterile field you need to throw away the vial.
All done!