Lesson 7: Test for Specified Microorganisms in Paracetamol Oral Suspe

Tests for Specified Microorganisms

Use specialized enrichment procedures to identify the absence or restricted presence of particular microorganisms

predetermined standard

Tests for Specified Microorganisms

1. Ensure that a substance or preparation meets a ____________________ for microbiological quality

2. Determination of the absence of, or limited occurrence of, specific ________________ that may be detected under the conditions described

1 = ?

microorganisms

Tests for Specified Microorganisms

1. Ensure that a substance or preparation meets a ____________________ for microbiological quality

2. Determination of the absence of, or limited occurrence of, specific ________________ that may be detected under the conditions described

2 = ?

antimicrobial properties

Make sure that before performing the method suitability tests, take note of the following:

1. If a product possesses _______________________________, these should be eliminated or neutralized.

2. If __________________ are used in sample preparation, non-toxicity toward the test organism and compatibility with any inactivators used must be demonstrated

1 = ?

surface-active agents

Make sure that before performing the method suitability tests, take note of the following:

1. If a product possesses _______________________________, these should be eliminated or neutralized.

2. If __________________ are used in sample preparation, non-toxicity toward the test organism and compatibility with any inactivators used must be demonstrated

1 = ?

Growth Promotion Test (GPT)

Used to test if the medium supports/allows the growth of organisms. In this case, it is used to test if Escherichia coli can grow in MacConkey Broth.

5 mL of MacConkey Broth

GPT PROCESS:

1. Place _________________ in two test tubes.

2. In the 1st test tube, introduce ____________________________________ and label it as GPT-Test.

3. In the 2nd test tube, only add the MCB and label it as _______________.

4. Pack the two tubes with __________________ and label with GPT, Incubation, Temperature, Section, Group Number, and Pull-out date. Incubate at 30°C - 35°C for 3 days.

5. Check for signs of _____________ on the medium.

1 = ?

0.1 mL of 90 cfu of Escherichia coli bacterial suspension

GPT PROCESS:

1. Place _________________ in two test tubes.

2. In the 1st test tube, introduce ____________________________________ and label it as GPT-Test.

3. In the 2nd test tube, only add the MCB and label it as _______________.

4. Pack the two tubes with __________________ and label with GPT, Incubation, Temperature, Section, Group Number, and Pull-out date. Incubate at 30°C - 35°C for 3 days.

5. Check for signs of _____________ on the medium.

2 = ?

GPT-Control

GPT PROCESS:

1. Place _________________ in two test tubes.

2. In the 1st test tube, introduce ____________________________________ and label it as GPT-Test.

3. In the 2nd test tube, only add the MCB and label it as _______________.

4. Pack the two tubes with __________________ and label with GPT, Incubation, Temperature, Section, Group Number, and Pull-out date. Incubate at 30°C - 35°C for 3 days.

5. Check for signs of _____________ on the medium.

3 = ?

aluminum foil

GPT PROCESS:

1. Place _________________ in two test tubes.

2. In the 1st test tube, introduce ____________________________________ and label it as GPT-Test.

3. In the 2nd test tube, only add the MCB and label it as _______________.

4. Pack the two tubes with __________________ and label with GPT, Incubation, Temperature, Section, Group Number, and Pull-out date. Incubate at 30°C - 35°C for 3 days.

5. Check for signs of _____________ on the medium.

4 = ?

microbial growth

GPT PROCESS:

1. Place _________________ in two test tubes.

2. In the 1st test tube, introduce ____________________________________ and label it as GPT-Test.

3. In the 2nd test tube, only add the MCB and label it as _______________.

4. Pack the two tubes with __________________ and label with GPT, Incubation, Temperature, Section, Group Number, and Pull-out date. Incubate at 30°C - 35°C for 3 days.

5. Check for signs of _____________ on the medium.

5 = ?

turbidity

ANTICIPATED RESULTS OF GROWTH PROMOTION TEST

a. Accepted Criteria: Clear, visible growth of the microorganisms (_________ is present in the test sample relative to the control)

b. Interpretation: Growth Promotion Test is used to determine if MacConkey Broth is suitable to detect _______________ in the Test for Specified Microorganisms

a = ?

Escherichia coli

ANTICIPATED RESULTS OF GROWTH PROMOTION TEST

a. Accepted Criteria: Clear, visible growth of the microorganisms (_________ is present in the test sample relative to the control)

b. Interpretation: Growth Promotion Test is used to determine if MacConkey Broth is suitable to detect _______________ in the Test for Specified Microorganisms

b = ?

Inhibitory Test

MacConkey Broth inhibits the growth of gram-positive bacteria, such as Staphylococcus aureus, because it contains bile salts and crystal violet.

5 mL of MacConkey Broth

INHIBITORY TEST PROCESS:

1. Place ______________________ in a test tube.

2. Inoculate the test tube with _______________________________________.

3. Cover with ___________ and label with Inhibitory Test, Incubation, Temperature, Section, Group number, and pull-out date. Incubate at 30°C - 35°C for 3 days.

4. Check for signals of ________________ (turbidity) on the medium

1 = ?

0.2 mL (180 cfu) of Staphylococcus aureus bacterial suspension

INHIBITORY TEST PROCESS:

1. Place ______________________ in a test tube.

2. Inoculate the test tube with _______________________________________.

3. Cover with ___________ and label with Inhibitory Test, Incubation, Temperature, Section, Group number, and pull-out date. Incubate at 30°C - 35°C for 3 days.

4. Check for signals of ________________ (turbidity) on the medium

2 = ?

aluminum foil

INHIBITORY TEST PROCESS:

1. Place ______________________ in a test tube.

2. Inoculate the test tube with _______________________________________.

3. Cover with ___________ and label with Inhibitory Test, Incubation, Temperature, Section, Group number, and pull-out date. Incubate at 30°C - 35°C for 3 days.

4. Check for signals of ________________ (turbidity) on the medium

3 = ?

microbial growth

INHIBITORY TEST PROCESS:

1. Place ______________________ in a test tube.

2. Inoculate the test tube with _______________________________________.

3. Cover with ___________ and label with Inhibitory Test, Incubation, Temperature, Section, Group number, and pull-out date. Incubate at 30°C - 35°C for 3 days.

4. Check for signals of ________________ (turbidity) on the medium

4 = ?

No growth

ANTICIPATED RESULTS OF INHIBITORY TEST:

a. Acceptance Criteria: __________ of test microorganisms should occur.

b. Interpretation: The inhibitory test is used to demonstrate the suitability of MacConkey Broth in ________________________________________ like Staphylococcus aureus.

a = ?

inhibiting the growth of gram-positive bacteria

ANTICIPATED RESULTS OF INHIBITORY TEST:

a. Acceptance Criteria: __________ of test microorganisms should occur.

b. Interpretation: The inhibitory test is used to demonstrate the suitability of MacConkey Broth in ________________________________________ like Staphylococcus aureus.

b = ?

Indicative Test

This test is used to test if the MacConkey Agar can differentiate between lactose fermenters and non-lactose fermenters. A pH indicator such as neutral red will turn pink/red if the grown bacteria are a lactose-fermenter, such as Escherichia coli. 

15-20 mL of MacConkey Agar

Process of Indicative Test:

1. Place approximately ________________________ (cooled to 45°C) into two sterile petri dishes and allow to solidify.

2. Allow the plates to dry in a ___________________________________.

3. Inoculate the 1st petri dish with ____________________________________ using the Spread plate method. Label as Indicative–Test.

4. In the 2nd petri dish with the MacConkey Agar, label it as ________________.

5. Pack the two plates with _____ and label with Indicative Test, Incubation Temperature, section, group number, and pull-out date. Incubate at 30°C - 35°C for 3 days.

6. __________ for changes in the medium and the presence of colonies.

1 = ?

laminar-airflow cabinet or an incubator

Process of Indicative Test:

1. Place approximately ________________________ (cooled to 45°C) into two sterile petri dishes and allow to solidify.

2. Allow the plates to dry in a ___________________________________.

3. Inoculate the 1st petri dish with ____________________________________ using the Spread plate method. Label as Indicative–Test.

4. In the 2nd petri dish with the MacConkey Agar, label it as ________________.

5. Pack the two plates with _____ and label with Indicative Test, Incubation Temperature, section, group number, and pull-out date. Incubate at 30°C - 35°C for 3 days.

6. __________ for changes in the medium and the presence of colonies.

2 = ?

0.1 mL (90 cfu) of Escherichia coli bacterial suspension

Process of Indicative Test:

1. Place approximately ________________________ (cooled to 45°C) into two sterile petri dishes and allow to solidify.

2. Allow the plates to dry in a ___________________________________.

3. Inoculate the 1st petri dish with ____________________________________ using the Spread plate method. Label as Indicative–Test.

4. In the 2nd petri dish with the MacConkey Agar, label it as ________________.

5. Pack the two plates with _____ and label with Indicative Test, Incubation Temperature, section, group number, and pull-out date. Incubate at 30°C - 35°C for 3 days.

6. __________ for changes in the medium and the presence of colonies.

3 = ?

Indicative–Control

Process of Indicative Test:

1. Place approximately ________________________ (cooled to 45°C) into two sterile petri dishes and allow to solidify.

2. Allow the plates to dry in a ___________________________________.

3. Inoculate the 1st petri dish with ____________________________________ using the Spread plate method. Label as Indicative–Test.

4. In the 2nd petri dish with the MacConkey Agar, label it as ________________.

5. Pack the two plates with _____ and label with Indicative Test, Incubation Temperature, section, group number, and pull-out date. Incubate at 30°C - 35°C for 3 days.

6. __________ for changes in the medium and the presence of colonies.

4 = ?

foil

Process of Indicative Test:

1. Place approximately ________________________ (cooled to 45°C) into two sterile petri dishes and allow to solidify.

2. Allow the plates to dry in a ___________________________________.

3. Inoculate the 1st petri dish with ____________________________________ using the Spread plate method. Label as Indicative–Test.

4. In the 2nd petri dish with the MacConkey Agar, label it as ________________.

5. Pack the two plates with _____ and label with Indicative Test, Incubation Temperature, section, group number, and pull-out date. Incubate at 30°C - 35°C for 3 days.

6. __________ for changes in the medium and the presence of colonies.

5 = ?

Check

Process of Indicative Test:

1. Place approximately ________________________ (cooled to 45°C) into two sterile petri dishes and allow to solidify.

2. Allow the plates to dry in a ___________________________________.

3. Inoculate the 1st petri dish with ____________________________________ using the Spread plate method. Label as Indicative–Test.

4. In the 2nd petri dish with the MacConkey Agar, label it as ________________.

5. Pack the two plates with _____ and label with Indicative Test, Incubation Temperature, section, group number, and pull-out date. Incubate at 30°C - 35°C for 3 days.

6. __________ for changes in the medium and the presence of colonies.

6 = ?

pink/red, and the medium also turns pink/red

ANTICIPATED RESULTS OF INDICATIVE TEST

a. Acceptance Criteria:

1. Colonies that are formed on the Petri dish with the test sample appear ___________________________.

2. The control should have no pink/red colonies, and the medium should remain __________.

b. Interpretation: An indicative test is used to demonstrate that the colonies formed on the MacConkey Agar with the characteristic appearance described are indicative of _______________\

a.1. = ?

unchanged

ANTICIPATED RESULTS OF INDICATIVE TEST

a. Acceptance Criteria:

1. Colonies that are formed on the Petri dish with the test sample appear ___________________________.

2. The control should have no pink/red colonies, and the medium should remain __________.

b. Interpretation: An indicative test is used to demonstrate that the colonies formed on the MacConkey Agar with the characteristic appearance described are indicative of _______________\

a.2. = ?

Escherichia coli

ANTICIPATED RESULTS OF INDICATIVE TEST

a. Acceptance Criteria:

1. Colonies that are formed on the Petri dish with the test sample appear ___________________________.

2. The control should have no pink/red colonies, and the medium should remain __________.

b. Interpretation: An indicative test is used to demonstrate that the colonies formed on the MacConkey Agar with the characteristic appearance described are indicative of _______________\

b = ?

paracetamol oral suspension

Suitability of Test Method

Preparation of the Diluted Test Product

1. Shake the bottle of __________________________ before opening.

2. Cleanse the outer surfaces of the container with ________________________.

3. _________ open the container.

4. Accurately measure and transfer _________________________ using a pipette into a beaker.

5. Add _______________ and mix.

1 = ?

sterile cotton swabs containing 70% ethyl alcohol

Suitability of Test Method

Preparation of the Diluted Test Product

1. Shake the bottle of __________________________ before opening.

2. Cleanse the outer surfaces of the container with ________________________.

3. _________ open the container.

4. Accurately measure and transfer _________________________ using a pipette into a beaker.

5. Add _______________ and mix.

2 = ?

Aseptically

Suitability of Test Method

Preparation of the Diluted Test Product

1. Shake the bottle of __________________________ before opening.

2. Cleanse the outer surfaces of the container with ________________________.

3. _________ open the container.

4. Accurately measure and transfer _________________________ using a pipette into a beaker.

5. Add _______________ and mix.

3 = ?

10 mL of the test product

Suitability of Test Method

Preparation of the Diluted Test Product

1. Shake the bottle of __________________________ before opening.

2. Cleanse the outer surfaces of the container with ________________________.

3. _________ open the container.

4. Accurately measure and transfer _________________________ using a pipette into a beaker.

5. Add _______________ and mix.

4 = ?

90 mL of SCDB

Suitability of Test Method

Preparation of the Diluted Test Product

1. Shake the bottle of __________________________ before opening.

2. Cleanse the outer surfaces of the container with ________________________.

3. _________ open the container.

4. Accurately measure and transfer _________________________ using a pipette into a beaker.

5. Add _______________ and mix.

5 = ?

10 mL of diluted test product

METHOD SUITABILITY TEST (METHOD VALIDATION):

Suitability of Test Method

1.Transfer ______________________________ into a test tube,

2.Add ___________________________________________. Mix.

3.Label the ______ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

4.Incubate at ____________________

5.Shake the container and transfer ______________________________________ in a beaker. Cover the beaker with aluminum foil.

6.Label the ________ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

7.Incubate at ____________________

8.Inoculate ________________________.

9.Pack with ___________ and label with Suitability Test, microorganism, incubation temperature, section and group

10.Incubate at ______________

1 = ?

0.1 mL (90 cfu) of the bacterial suspension of E. coli

METHOD SUITABILITY TEST (METHOD VALIDATION):

Suitability of Test Method

1.Transfer ______________________________ into a test tube,

2.Add ___________________________________________. Mix.

3.Label the ______ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

4.Incubate at ____________________

5.Shake the container and transfer ______________________________________ in a beaker. Cover the beaker with aluminum foil.

6.Label the ________ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

7.Incubate at ____________________

8.Inoculate ________________________.

9.Pack with ___________ and label with Suitability Test, microorganism, incubation temperature, section and group

10.Incubate at ______________

2 = ?

tube

METHOD SUITABILITY TEST (METHOD VALIDATION):

Suitability of Test Method

1.Transfer ______________________________ into a test tube,

2.Add ___________________________________________. Mix.

3.Label the ______ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

4.Incubate at ____________________

5.Shake the container and transfer ______________________________________ in a beaker. Cover the beaker with aluminum foil.

6.Label the ________ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

7.Incubate at ____________________

8.Inoculate ________________________.

9.Pack with ___________ and label with Suitability Test, microorganism, incubation temperature, section and group

10.Incubate at ______________

3 = ?

30-35 C for 18-24 hours

METHOD SUITABILITY TEST (METHOD VALIDATION):

Suitability of Test Method

1.Transfer ______________________________ into a test tube,

2.Add ___________________________________________. Mix.

3.Label the ______ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

4.Incubate at ____________________

5.Shake the container and transfer ______________________________________ in a beaker. Cover the beaker with aluminum foil.

6.Label the ________ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

7.Incubate at ____________________

8.Inoculate ________________________.

9.Pack with ___________ and label with Suitability Test, microorganism, incubation temperature, section and group

10.Incubate at ______________

4 = ?

1 mL of the contents to 100 mL of MacConkey Broth

METHOD SUITABILITY TEST (METHOD VALIDATION):

Suitability of Test Method

1.Transfer ______________________________ into a test tube,

2.Add ___________________________________________. Mix.

3.Label the ______ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

4.Incubate at ____________________

5.Shake the container and transfer ______________________________________ in a beaker. Cover the beaker with aluminum foil.

6.Label the ________ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

7.Incubate at ____________________

8.Inoculate ________________________.

9.Pack with ___________ and label with Suitability Test, microorganism, incubation temperature, section and group

10.Incubate at ______________

5 = ?

beaker

METHOD SUITABILITY TEST (METHOD VALIDATION):

Suitability of Test Method

1.Transfer ______________________________ into a test tube,

2.Add ___________________________________________. Mix.

3.Label the ______ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

4.Incubate at ____________________

5.Shake the container and transfer ______________________________________ in a beaker. Cover the beaker with aluminum foil.

6.Label the ________ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

7.Incubate at ____________________

8.Inoculate ________________________.

9.Pack with ___________ and label with Suitability Test, microorganism, incubation temperature, section and group

10.Incubate at ______________

6 = ?

42-44 C for 24 hours

METHOD SUITABILITY TEST (METHOD VALIDATION):

Suitability of Test Method

1.Transfer ______________________________ into a test tube,

2.Add ___________________________________________. Mix.

3.Label the ______ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

4.Incubate at ____________________

5.Shake the container and transfer ______________________________________ in a beaker. Cover the beaker with aluminum foil.

6.Label the ________ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

7.Incubate at ____________________

8.Inoculate ________________________.

9.Pack with ___________ and label with Suitability Test, microorganism, incubation temperature, section and group

10.Incubate at ______________

7 = ?

0.1 mL on MacConkey Agar

METHOD SUITABILITY TEST (METHOD VALIDATION):

Suitability of Test Method

1.Transfer ______________________________ into a test tube,

2.Add ___________________________________________. Mix.

3.Label the ______ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

4.Incubate at ____________________

5.Shake the container and transfer ______________________________________ in a beaker. Cover the beaker with aluminum foil.

6.Label the ________ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

7.Incubate at ____________________

8.Inoculate ________________________.

9.Pack with ___________ and label with Suitability Test, microorganism, incubation temperature, section and group

10.Incubate at ______________

8 = ?

aluminum foil

METHOD SUITABILITY TEST (METHOD VALIDATION):

Suitability of Test Method

1.Transfer ______________________________ into a test tube,

2.Add ___________________________________________. Mix.

3.Label the ______ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

4.Incubate at ____________________

5.Shake the container and transfer ______________________________________ in a beaker. Cover the beaker with aluminum foil.

6.Label the ________ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

7.Incubate at ____________________

8.Inoculate ________________________.

9.Pack with ___________ and label with Suitability Test, microorganism, incubation temperature, section and group

10.Incubate at ______________

9 = ?

30-35 C for 18 hours

METHOD SUITABILITY TEST (METHOD VALIDATION):

Suitability of Test Method

1.Transfer ______________________________ into a test tube,

2.Add ___________________________________________. Mix.

3.Label the ______ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

4.Incubate at ____________________

5.Shake the container and transfer ______________________________________ in a beaker. Cover the beaker with aluminum foil.

6.Label the ________ with Suitability Test, microorganism, incubation temperature, section and group number, and the pull-out date.

7.Incubate at ____________________

8.Inoculate ________________________.

9.Pack with ___________ and label with Suitability Test, microorganism, incubation temperature, section and group

10.Incubate at ______________

10 = ?

Colonies

Suitability of Test Method:

a. Acceptance Criteria: _______________ formed on the Petri dish with the test sample appear pink/red and the medium also turns into pink/red.

b. Interpretation: Suitability of Test Method demonstrates that method is _________ of Escherichia coli in the drug product.

a = ?

suitable to detect the presence

Suitability of Test Method:

a. Acceptance Criteria: _______________ formed on the Petri dish with the test sample appear pink/red and the medium also turns into pink/red.

b. Interpretation: Suitability of Test Method demonstrates that method is _________ of Escherichia coli in the drug product.

b = ?

Suitability of Test Method

Product Testing has the same procedure as _______________ except do not inoculate E. coli.

Absence of pink/red colonies

PRODUCT TESTING:

a. Acceptance Criteria: _____________________________ on the plated medium.

b. Interpretation: The test method will determine the ________________________ in the drug product.

a = ?

presence or absence of Escherichia coli

PRODUCT TESTING:

a. Acceptance Criteria: _____________________________ on the plated medium.

b. Interpretation: The test method will determine the ________________________ in the drug product.

b = ?