Test Base into water aka Suspension
1 - Large Pyrex glass flask (sealable is preferred)
Polysorbate 80 (.5% only)
1 – large Pyrex glass mixing cup (you can get this at Dillon’s)
Latex gloves (recommended for sterility)
1 - Small Stainless steel whisk (the kind for whipping eggs that you get at Dillon’s)
1 – small plastic funnel
Step 1.) MAKING BACTERIOSTATIC WATER.
1. Put distilled water into a pan and bring to a boil for 2 minutes.
2. Now add ice to bring water to a cold temp as quickly as possible.
3. Add 1% Benzyl Alcohol and .25% PolySorbate 80 to sterile container **percentages need to be in proper ratio to total volume of liquid**
4. Make sure all ice is melted, and then filter water into large sterile container where we added the BA and PS80 (this is where we will store our “bulk” bacteriostatic water.)
5. Now swirl the mixture so that it is totally absorbed, and now our bacteriostatic water is finished.
Step 2.) STERALIZING YOUR EQUIPMENT.
1. Fill 500ml Pyrex glass mixing cup with 90% isopropyl alcohol (not 70%) and put egg whisk in the mixing cup.
2. Let egg whisk and mixing cup soak for 30 minutes.
3. Now set aside the whisk and using your funnel, pour your alcohol back into the container (the alcohol can be reused).
4. Let both the whisk and the mixing cup air dry.
5. **This same method is great for sterilizing vials, however after they air dry it is good to bake them at 300 Deg. For 15 minutes.**
Step 3.) PREPARING YOUR TEST SUSPENTION.
1. Add 10% Benzyl Benzoate and .25% PS80 (that is 10% of total volume) **i.e. if you are making 100mls of Test Suspension then add 10ml of Benzyl Benzoate and .25ml of PS80.
2. Using a digital scale weigh out the proper amount of Test Base BP98 powder. (I usually make mine at 50mg/ml.) Example: for 50mg/ml use this formula, (total mls X .050) or (100mls X .050 = 5 Grams of Test Powder)
3. Add the Test powder to the Benzyl Benzoate and let soak for about 5 minutes, stirring gently with the egg whisk.
4. After the powder is totally saturated and there are no visible clumps, it is time to add your bacteriostatic water using the following formula: BB + (Powder X .75) +/- + Total volume = Water needed. **i.e. 10ml BB +(5Grams X .75) = 13.75 +/- +100mls = 86.25 mls of bacteriostatic water needed.
5. Now when you add your bacteriostatic water the BB/TNE mix will clump up at the bottom of the mixing cup, this is normal.
6. Put your mixing cup in a saucepan on your oven and turn burner to medium heat.
7. Slowly swirl your mixture as it heats, this will cause the clumps of BB/TNE mixture to disperse out into the water.
8. When no more clumps are visible the suspension is complete (it should be steaming slightly by now).
9. Remove from heat, draw into a large plunger and fill your vials. (Swirl your mixture with the whisk as you do this.)
10. Now add stoppers, seals, crimp and label. Whalah! You’re done!!
This formula was 6 months in the making, and I would like to thank all of you for your moral support and encouragement. I hope you will like the results. I personally guarantee that if done right this formula will not burn, cause redness and swelling, and will pass easily through a 30ga. Insulin pin. With no waste “clumps” at the bottom of a vial when it is used.