Page 23 - Laboratory manual for students FAR222 2019 20

FAR 222 Dosage Form II Laboratory Manual

EXPERIMENT 2: Sterility Test for Ampoule Sample

1. For this experiment, you are given: 2 sterile test tubes, 1 plate agar and 1 syringe with
needle.
2. Using a sterile pipette, aseptically dispense 8 mL of nutrient broth to each of the test tube.
Label the tubes as No.1 and 2.
3. Before you proceed to breaking open the glass ampoule, wash your hands thoroughly
first.
4. If there is liquid at the upper tip of the ampoule (above the breakage line), gently tap the
ampoule with your finger to get all the liquid to the bottom part of the ampoule.
5. Protect your hands from the broken glass by using paper towel or cotton wool when
opening the ampoule.
6. To open the ampoule, hold it with both hands, with one thumb against the narrow top
section. Hold the bottom of the ampoule firmly while pushing the top section away from
you with easy, even pressure (A light pressure should cleanly snap the ampoule open,
while using too much force can cause it to shatter).
7. Using a sterile syringe/needle, transfer 2 mL of the ampoule contents into test tube No. 1.
Allow the mixture to mix well, and transfer 2 mL of the mixture from test tube No. 1 into
test tube No. 2.
8. Take an agar plate and divide the plate into 2 sectors, mark each sector as 1 and 2.
9. Using a sterile wire loop, take a loopful of the dilution and streak on each sector
correspondingly.
10. Incubate your sample at 37°C, overnight (this includes both the serial dilution test tubes
and your inoculated plate agar).
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EXPERIMENT 3: Sterility Test for Unknown Sample

1. For this experiment, you are given: 2 sterile test tubes and 1 plate agar.
2. Using a sterile pipette, aseptically dispense 5 mL of nutrient broth to each of the test tube.
Label the tubes as No.1 and 2.
3. Using a sterile pipette, transfer 5 mL of unknown sample to test tube No. 1. Mix gently.
4. Subsequently, using a sterile pipette, transfer 5 mL of the broth/sample mixture from test
tube No. 1 to test tube No. 2.
5. Take an agar plate and divide the plate into 2 sectors, mark each sector as 1 and 2.
6. Using a sterile wire loop, take a loopful of the dilution and streak on each sector
correspondingly.
7. Incubate your sample at 37°C, overnight (this includes both the serial dilution test tubes
and your inoculated plate agar).

BEFORE LEAVING THE LAB:
Arrange your test tubes on the racks provided in this order:
4 (VIAL) 2 (AMPOULE) 2 (UNKNOWN)
3 plates, inverted

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