A workshop with limited number of participants

1. Sterile Operations for Parenteral Preparations
(Tutorial-6-1)
Roohi B. Obaid
23 Jun 2018 at Karachi
[151 Slides]

2. Fundamental Essentials
&
Regulatory Obligations

3. Discussion
• Science & good
science
Sharing
• Current
Knowledge &
Experience
Navigation
• Towards mutual
goal & Future
directions
Knowledge Sharing Exercise
To Learn through Debate, Discussion & Experience of others
Acknowledgement: Documents of US-FDA & PDA consulted

4. Pharmaceutical Microbiology

5. Essentials, Fundamentals, Building
Blocks & its Coordinated
Applications
Discussion on Several Real Time
Scenarios & Phenomena
Three (3) Challenging Cases of
Hardliner Atypical Possibilities
1
2
3

7. What will happen if an
injection is manufactured
in an environment where
tablet is manufactured
1: ………………..
2: ………………..
3: ………………..
4: ………………..
..
..

8. Infections and their load on Health Care

9. Moist Heat Sterilizer

10. Moist Heat Sterilizer
1: ………………..
2: ………………..
3: ………………..
4: ………………..
..
..

11. Recall the clinics of 80s

12. What happened
?

13. Certainty
Confidence
Trust Predictibility

14. Dry Heat Sterilizer

15. Dry Heat Sterilizer
1: ………………..
2: ………………..
3: ………………..
4: ………………..
..
..

16. Sterilization through
Filtration

17. Sterilization through
Filtration
1: ………………..
2: ………………..
3: ………………..
4: ………………..
..
..

18. Sterilization through
Radiation

19. Sterilization through
Radiation
1: ………………..
2: ………………..
3: ………………..
4: ………………..
..
..

21. What next
?

22. A- Septic Area

23. Certainty
Confidence
Trust Predictability

24. Challenge … Media Fill …. Qualify ….

25. How would you clean
the area after media
filling test
?

26. Again
Its you to think, share and lets discuss to see where we are?

27. When life is everything
Risk

28. P A R E N T E R A L S
IV
IM
IG
SC
.
.
.

29. Horrible sometimes …

30. Clinical Effects of Injected Particulate Matter
Phlebitis
Pulmonary
emboli
Pulmonary
granulomas
Immune
system
dysfunction
Pulmonary
dysfunction
Infarction Death

32. Prokaryotes Eukaryotes

36. Antony van Leeuwenhook
(Dutch)
1632 - 1723

38. Life Life
With
or
Without

39. Biogenesis
or
A-Biogenesis

40. Franseco Redi
(Italy)
1626 - 1697

41. Redi
?

42. What is the potential question?

43. Absence of air cannot promote growth
Theory challenged

44. Louis Pasteur
(French)
1822 - 1895

45. Louis Pasteur
?

46. What is the potential question?

47. Nothing left unattended, O2, etc.
Life from Life

48. Hans Christian Gram
(Danish)
1853 - 1938

54. Gram positive
Gram negative

55. Gram positive
Gram negative
Acid fast

56. Strepto
Staphylo
Diplo / pair

57. Aerobe
Anaerobe
Facultative

58. Aerobe
Anaerobe
Facultative aerobe
Gram
Positive
Gram
Negative

59. Stain
Colony
Shape Oxygen

60. Tiny living
things
Microorganisms that exist as
single cells capable of growth,
respiration, reproduction
independent of other cells of the
same or different kind.

61. G
R
O
W
T
H

62. Cannot swim itself
Cannot move itself
Good Luck

63. Good Luck
Sensitive to heat
Sensitive to radiation
Sensitive to chemicals

64. Aerobe
Facultative aerobe
Anaerobe
O2
No
O2
O2 or no O2

65. Spore formation
Thermophilic
(40-70℃)
Mesophilic
(15-45℃)
Psychrophilic
(0-20℃)
Challenges

66. Volcanic
Vents
• High
temperature
Antarctica
• Low
temperature
• Low
nutrients
Mount
Everest
• O2, low
temperature,
low nutrients
Sahara
Desert
• low moisture,
high
temperature,
low nutrients
WFI
Systems
• Low
nutrients
Challenges (Extremes)

67. Dormant, tough,
non-reproductive
structure
Forms in response
to environmental
stress
Can survive for
extended periods
of time
Produced by
Gram Positive (+)
Bacillus (rod)
Challenges (Endospores)

68. Water borne
Air borne
Human
Challenges

69. Skill to handle
Knowledge
Ability/ Aptitude
Challenges

70. From where vulnerability begins
Nutrients Respiration Temperature Humidity pH
Water also carry the microorganisms
and supports growth too

71. Putting it all to use
Cocci, Gram +ve
Small/
medium
colony
White,
beige to
yellow
Circular
and
convex
Buttery
Human
Operation/ skin/
hands & hair
Staphylococcus
species
1

72. Putting it all to use
Rods, Gram +ve
Medium/
large
colony
Off white
to beige
Circular/
irregular
and
convex/
raised
Mucoid,
dry to
grainy
Environment such
as soil (seasonal
factor)
Through shoe, air
etc.
Clostridium
species
2

73. Putting it all to use
Rods, Gram -ve
Small to
medium
colony
White,
off
white to
beige
Circular/
irregular
and
convex
Mucoid
to
buttery
Generally found in
water (equipment
piping )
Pseudomonas, E.
coli species
3

74. Staphylococcus
aureus
• Aerobic
• Gram +ve
• Coccus
Pseudomonas
aeruginosa
• Aerobic
• Gram -ve
• Rod
Bacillus subtilis
• Aerobic
• Gram +ve
• Rod
• Spore forming
Clostridium
sporogenese
• Anaerobic
• Gram +ve
• Rod
• Spore forming
Candida albicans
• Yeast
Aspergillus
brasiliensis
• Mold
Compendial Organisms USP …ICH

75. Horrible sometimes …

76. Lets take another view
Non-viable particulate matters in Parenterals

77. Sources of Particulate Matter in Injectable Drug Products
Environment
Packaging
materials
Solution &
formulation
components
Product-
packaging
interactions
Process
generated
particles

78. Take an area as an example of challenge

79. 2m2
• Protects body from
pathogens
• Prevents loss of
moisture
• Body temperature
regulation
Diversity
• Considerable diversity
in species
• Variation b/w different
locations on body
• Variation b/w
individuals
30 million
• Average sheds up
to 1 billion skin
cells/ day
• 10% have micro-
organisms on them
Human Skin
Go in clean rooms They touch, shed, do things wrong sometimes

80. ORAL CAVITY
High population
High diversity
SKIN
Variable population
Low diversity
URETHRA/VAGINA
High population
Low diversity
GUT
High population
High diversity
People
Ecology

81. Patient Population
Patients with
existing tissue
damage e.g. trauma,
surgery or sepsis
Critically ill patients Neonates
At highest
risk

82. Sterility: complete absence of life or
inability to reproduce
• Cont’d
In pharmaceutical industry ?

83. Sterility: More Complex & Technical
• Cont’d
Sterility: Expressed in Quantitative
Terms

84. The probability definition for sterility derives
from the fact that microbial death follows a
geometric progression .i.e. the cells do not all
die at once when exposed to lethal conditions
• Cont’d
The US-FDA defines sterilization as the
“reduction of a microbial population to 10° (or
one cell) plus an additional 6 log-cycles”

85. Sterility : Probability of a non-sterile unit in a lot or
Sterility Assurance Level
A product is considered sterile when the probability of a
non-sterile unit is < 1 in 1, 000, 000 units; or a SAL of
10-6

86. Microorganis
ms die at a
constant rate
over a period
of time
For e.g. one
million
bacteria with
a 90% death
rate/ min
1st min
9 Lac
dead/ 1 lac
survivors
2nd min
90, 000 dead/
10, 000
survivors
Constant set of lethal conditions develop resistance
3rd min
9, 000
dead/
1, 000
survivors
4th min
900 dead/
100
survivors
5th min
90 dead/
10
survivors
6th min
9 dead/ 1
survivor
7th min
• ?

87. Human Being
Flaws in
Maintenance
Design &
Facilities

88. Recap & Lets apply

89. BBB?
What is BBB?
Can a bacteria cross BBB?
Can every drug cross BBB?
CONSEQUENCES?

90. Meningitis
Tragedy? An outbreak of meningitis in US (2012)
Poor aseptic practices in Compounding
Pharmacies
Methyl prednisolone was contaminated
64 patients died
753 affected

91. Meningitis
Tragedy?
Shipped product prior to result of
sterility
Prioritized production over cleaning
Forgery in cleaning log
64 patients died
753 affected

92. Meningitis
Tragedy?
Ignored molds & bacteria found inside
the clean room
Working by unqualified persons
It is reckless & disregard to profession
64 patients died
753 affected

93. Meningitis
Tragedy? Drug is required to be
manufactured under high
quality conditions so that NO
patient is at risk of harm due to
…..
Pay regard to patients safety
over everything
Lesson learned

94. Public Health & Safety, Take a Scenario for relative narration

95. Suppose an endoscope
device is contaminated
during cleaning or
manufacturing
Lets make consequences visible

96. 50, 000 to 100, 000 may be more
A rough estimate
Sometimes it is difficult to disinfect before
re-use
Multi drug resistant organisms?
Endoscope is inserted in the GIT

97. Particulate Matter

98. Sources of Particulate Matter in Injectable Drug Products
Environment
Packaging
materials
Solution &
formulation
components
Product-
packaging
interactions
Process
generated
particles

99. Clinical Effects of Injected Particulate Matter
Phlebitis
Pulmonary
emboli
Pulmonary
granulomas
Immune
system
dysfunction
Pulmonary
dysfunction
Infarction Death

100. Group Discussion
on Scenarios

101. Scenario
Two healthy children with minor RTA administered TT and died
one after one within six hours …. ?

102. Scenario
36 deaths in early 2000
Endotoxin contaminated IV (WFI & Glucose) was administered
Roraima, Brazil

103. Ampoules or empty vials before
filling or after filling handled in
loose flexible plastic bag
Is it
acceptable?

104. What would be your response for
sterile area?
Earthquake is
experienced?

105. Any construction or unstudied
vibration in adjacent area
connected with beams?
What should
be done?

106. A giant corporate story of careless
destruction became reason of
regulatory action
Lets see what
is the case

107. Regulatory Definitions

108. Airlock
Small rooms with
Air pressure control
b/w adjoining rooms
Diff. cleanliness level
Interlocked doors

109. Unidirectional Flow
Air flow moving in a single direction
In a robust & uniform manner
With sufficient speed to reproducibly sweep
particles away from the critical processing area

110. Alert Level
That triggers alarm
Address potential problem
That triggers action
Investigate & Identify root cause
Action Level

111. Laminar Flow
Moving in a single direction in parallel layers
At constant velocity from
beginning to end of a straight line vector

112. Turbulent Flow

113. D - Value
90%
Reduction in
population
XX
minutes of heat
exposure at given
temperature
1 log

114. Over kill Sterilization
A process that is sufficient to provide at least 12 log
reduction of microorganisms with min. D value of 1 minute
12
log reduction
D value is
1 min

115. 1 log reduction 100 90% 10

116. 1 log reduction 100 90% 10
2 log reduction 10 90% 1

117. 1 log reduction 100 90% 10
2 log reduction 10 90% 1
3 log reduction 1 90% 0.1

118. 1 log reduction 100 90% 10
2 log reduction 10 90% 1
3 log reduction 1 90% 0.1
4 log reduction 0.1 90% 0.01

119. 1 log reduction 100 90% 10
2 log reduction 10 90% 1
3 log reduction 1 90% 0.1
4 log reduction 0.1 90% 0.01
6 log reduction 0.001 90% ???

120. 1 log reduction 100 90% 10
2 log reduction 10 90% 1
3 log reduction 1 90% 0.1
4 log reduction 0.1 90% 0.01
6 log reduction 0.001 90% ???
12 log reduction ??? 90% ???

121. Gowning Qualification
Program qualifies capability to wear
gowning in an aseptic manner
Initially Periodically

122. Disinfection
Reduction in surface
bio-burden
Vegetative
Spore

123. Decontamination
Eliminate viable bio-burden
Sporicidal

124. De-pyrogenation
Destroy / remove pyrogen
Endotoxin

125. HEPA Filter
0.3 um
size minimum
99.97%
Retaining efficiency

126. HEPA Filter
10000 9997 3
100000 99970 30

127. ULPA Filter
0.3 um
size minimum
99.999%
Retaining efficiency

128. ULPA Filter
100000 99999 1
1000000 999990 10

129. Colony Forming Unit

130. Worst Case Scenario
Strict Process
Not so strict process
Lowest Chance
Highest Chance
Within SOP ... Failure chance?

131. Worst Case Scenario
Conditions
Upper limits, circumstances
that pose greatest chance of
failure either product or
process

132. Worst Case Scenario
It is proven acceptable range … in which
performance will not impact on attributes
while operating at the extremes of
proposed ranges

133. Air Classification
Particle size Microorganisms CFU
1 2

134. Air Classification
Particle size Microorganisms CFU
1 2
> 0.5 um/m3 ActivePlate

135. Air Classification
> 0.5 um/m3 ActivePlate

136. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1

137. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7

138. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10

139. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10
3520000 50 100

140. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10
3520000 50 100
Class

141. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10
3520000 50 100
100
Class

142. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10
3520000 50 100
100
1000
Class

143. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10
3520000 50 100
100
1000
10,000
Class

144. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10
3520000 50 100
100
1000
10,000
100,000
Class

145. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10
3520000 50 100
100
1000
10,000
100,000
ISOClass

146. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10
3520000 50 100
100
1000
10,000
100,000
ISO
5
Class

147. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10
3520000 50 100
100
1000
10,000
100,000
ISO
5
6
Class

148. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10
3520000 50 100
100
1000
10,000
100,000
ISO
5
6
7
Class

149. Air Classification
> 0.5 um/m3 ActivePlate
3520 1 1
35200 3 7
352000 5 10
3520000 50 100
100
1000
10,000
100,000
ISO
5
6
7
8
Class

150. Count of filling table found elevated …
Sample was done two days before …
What and how would you deal with it ..
Share any one reason of it too

151. Lunch Break …
Cont’d with Part 2 (2A, 2B & 2C)