I just made some sust and it filtered very very quickly with my peristaltic pump and polycap 36 as filter. A week later it has all gone cloudy. around 600ml of it. It was clear to start.
I believe this may be because it was filtered too quickly (i think in a few mins)
My question.. What PSI/Speed should the peristaltic pump be filtering the solution at... and if so.. what peristaltic pump should i be looking at getting.
At the moment i got the dolphin 75 from RLS At the moment i'm doing the following;
I have an air filter in the kitchen to keep the air clean. All the sides are wiped. I handle everything with gloves. I do everything on the kitchen work top which is where i do the crimping/bottling(with a bottletop dispenser)
1. I brew the gear using the peristaltic pump which takes one tube from the solution (ptfe) through the peristaltic tubing, straight into the polycap (with the little air outlet lid screwed on... perhaps thats supposed to be unscrewed?).... 2. I clean everything, put everything into the dishwasher... rinse with distilled water, then i put vials and stoppers into the microwave oven (using the convector oven setting to dry them.(any ideas how long it should be in there?) (about to purchase a dry heat steriliser for that part) 3. The solution at this point is in a media bottle screwed closed. I put the bottle top dispenser on a media bottle full of benzyl alcohol, i press a few times to purge, then i push a few more times to get the excess out. 4. i place the bottletop dispensor on top of the media bottle with the solution inside. 5. i dispense into the vial and put the rubber stopped in.. i do this for each vial (in the past i werent drying them in the microwave oven i was just taking them out of the dishwasher and putting the stoppers into the vials, this meant sometimes there was still a bit of water but i always flicked the water out before hand... and before the dishwasher i was using an autoclave so sometimes there was water still in the vials from the autoclave.. question here... is this water safe to be in that vial.. or what do you suggest to dry the vials.. will the dry heat steriliser be okay 6. at this point i put all the caps on the vials.. these up until i got the oven were also a little bit wet but i flicked the water out; a. i do have isopropyl alcohol to rinse them out with if you suggest. b. i do have plenty of benzyl to rinse with
Question; should i rinse everything with benzyl or IPA before i put it into the microwave or should i dishwasher everything then put everything into the microwave oven.. and what about the caps.
7. at this point i put everything under a uvc lamp/light steriliser and leave on for 20-30 minutes.
Can you please correct this process.. or suggest better ways for keeping everything sterile or sanitised. And can you tell me how to improve my processes and recommend pump etc.. i want everything to be perfect.. I plant to get a laminar flow hood in a week or so.
Also if i can contact you somehow i can show you the recipes and you can comment.. there are about 48 recipes i think alltogether i have compiled
just need to get it all perfect really.. please get back to me or pm me w/ever thanks guys
He knows what he's doing he just needs to fine tune his method. This happens quite often in a laboratory setting and when issues come about you talk with your colleagues about possible causes of problems and how to fix them.
He is here looking for some advice or for us to something he may have overlooked in his procedure that could be causing this issue.
I think to sterilize your vials you can give them a nice ethanol rinse then bake them in the oven for about 20 minutes. The dishwasher seems unnecessary.
Buddy, if you have all of the equipment as stated, but don't know what you're doing, you are over your head. Seriously, step back and think about it.
I have received little bits and pieces of advice from everyone. I support the idea of "buy cheap, buy twice" id rather get something that requires minimal effort and then learn how to use it 100% correctly. The only difference with syringe filters, vacuum filtration to polystaltic filtering is time and effort.
All i need to know is generally
1. the easiest / best (or both) method of making sure everything is entirely sterile without a laminar flow hood and then the different process with a laminar flow hood. And the equipment i have to do this is; IPA, BA, A dishwasher, An Autoclave, A Dry Heat Sterilizer (just won it on ebay), hydrogen peroxide solution, distilled water, commercial soap (not sure if this one leaves a residue) and a UVC Light 30w Bulb (remote control), a whole room HEPA Air Filter, nitryl gloves, a lab coat, and face masks, replacement peristaltic tubing, replacement In / Out tubing to go inside the peristaltic tubing.
2.the easiest / best way to use my equipment to the best of its ability; i have been told whatman polycap 36's can do 5l a time, and between compounds to just purge with Grapeseed Oil (i think purging with BA / IPA could of caused an issue as this could of cause some to still be in the tube
maybe im overcomplicating things, but i just wanna make the most of all the equipment i have, if some steps are redundant i would like someone to tell me so i can formulate my own guide. I will hapilly rewrite it and share it with forum members once i have adapted it. Once i have perfected the method i will be testing my blends and recipes and making sure everyone works perfect then i will incorporate those into a guide. I aim to by the end of it make a comprehensive guide (for my own personal reference) that i will probably share with everyone once complete. At the moment it includes where to get filters, where to get BA, BB, EO, Guaiacol, Polysorbate 80, GSO, from in the UK. (This information is very handy for UK members), about 8 or 9 different sterilisation and filtering methods lol.
I just wanna condense everything into 1. If you guys are happy i can paste all of the methods i have collected and you could direct me to which is the best one out of all. Or if you think its too much info i wont bother. Thanks to everyone who has serious intentions of helping me to get sorted.