Professional Documents
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VALIDATION OF
STERILIZATION EQUIPMENTS
Aseptic Area Validations
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Content:
Definition of Sterilization and Depyrogenation
Microbiological aspects of Sterilization and
Depyrogenation, Lethality calculation,
-D Value, FH & F0 Values
-Z Value and use of microbiological indicators.
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Content:
- Dry Heat Ovens
- Dry Heat Sterilization Tunnels
- Steam Sterilizator (Autoclaves)
1. Design Qualification
2. Installation Qualification
3. Operational Qualification
4. Performance Qualification
4.1. Thermodynamical aspects of Sterilization
3.2. Temperature Distribution and Heat
Penetration studies.
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Definitions:
1. Sterilization:
Validated process used to render a
product free of living microorganisms
including bacterial endospores.
2. Depyrogenation:
Removal or inactivation of bacterial
endotoxin.
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Sterilization Only:
The cycle is designed to assure that the
probability of survival of the native
microflora is no greater than one cell in one
million units of the commodity.
(10-6 probability of nonsterility)
Dry Heat Sterilization, Theoretical
requirement: 170 0C, 32 min.
Steam Sterilization Theoretical
requirement: 121 0C, 15 min.
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Sterilization - Overkill
The overkill approach provides
assurance of sterilization well in excess
of the 10-6 probability of non
- sterility.
For example an FH provided by an
overkill cycle may produce a 12
- log
reduction of a biological indicator that
exhibits a high resistance to dry heat.
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0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Minutes at Base Temp.
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Determination of Z - Value:
Determine the D- value of an organism
at min. three different temperatures.
Construct a Thermal Death Curve by
plotting the logarithm of the -D Value
versus temperature.
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Log D Value
100
D130 0C : 10 min.
Z Value = 20 0C
10
130
140
150
0,1
Temperature 0C
160
170
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Z-Value:
In general, for Dry Heat sterilization,
- Value may be assumed as 20 0C.
Z
And for Steam Sterilization as 10 0C.
However, it will be appropriate to verify
for the biological indicators when they are
used to measure the integrated lethality
of a dry heat or steam sterilization cycle.
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LETHALITY RATE:
Also defined as :
FH For Dry Heat Sterilization
Fo For Steam Sterilization
The equivalent sterilization time spent
at the base temperature.
Tb : 170 oC (For Dry Heat Sterilization)
Tb : 121 oC (For Steam Sterilization)
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LETHALITY CALCULATION
Patashnik Method
Lethality Rate : 10
t
T
Tb
Z
(T-Tb)/Z
FH = t x Lethality Rate
: Cycle time
: Actual Cycle temperature
: Base Temperature
: Microbial Death Rate Constant
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LETHALITY CALCULATION
Example:
Determination of FH of a 3 min. dry heat
sterilization cycle at 175 0C
t = 3 min
T = 175 0C
FH = 4 x 10 (175-170)/20
Tb = 170 0C
FH = 5.31
z = 20 0C
Sterilization at 175 0C for 3 min. is equivalent
to 5.31 min. at 170 0C .
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Temperature
(0C)
105
110
120
135
150
165
170
172
174
174
174
175
165
150
140
130
110
105
of Lethal Rates :
10.1912
FH = t x of Lethal
Rates
t = 5 min.
FH = 5 x 10.192
FH = 50.961 min. at
170 0C .
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PART-1
DRY HEAT
STERILIZATION
AND
DEPYROGENATION VALIDATION
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DRY HEAT
STERILIZATION & DEPYROGENATION
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Thermodynamical Aspects of
Heating Process:
1. Convection Heating Process:
The heat transfer through a medium
by motion of it s parts. Natural
convection is a result of differences in
density caused by temperature
gradients in the fluid mass.
Forced convection heating is effected
by the action of a mechanical device.
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Thermodynamical Aspects of
Heating Process:
2. Conduction Heating Process:
Conduction is accomplished ether by a
molecular interaction from higher energy
level to a lower energy level or by free
electrons.
Thus, the ability of solids to conduct heat
varies directly with the free electron
concentration. Pure metals are best
conductors and non
- metals are the poorest.
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Thermodynamical Aspects of
Heating Process:
3. Radiant Heating Process:
Radiant heating is the process which
energy flows from high temperature
body to a lower temperature.
The geometry of both source and the
exposure section of the unit will affect
the uniformity of the radiation density
in a unit.
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Manuel Transfer
Filling
Manuel Transfer
1. CONTINUOUS PROCESSING:
Washing
Washing
Sterilization
(Tunnel)
Continuous Transfer
Filling
Filling
Continuous Transfer
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Tunnel Sterilizer
Pressure Differential
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Temperature Distribution:
External monitoring and recording instruments
shall be calibrated before and after the OQ/PQ
studies (3 point calibration, 0.5 0C tolerance).
Uniformity of the temperature distribution in
case of Min&Max. loading should be verified by
using Thermocouples with 3 replicates.
T/C (Thermocouple) placement shall be
documented on a diagram.
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Temperature Distribution:
Min. 10-12 T/C s shall be used and they
should not be inserted in the load. Data should
be recorded during the whole cycle at 1 min.
intervals.
At least one T/C shall be placed adjacent to the
equipment temperature controller.
Location of the cold spot should be
determined and documented.
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S7
b7
S4
7
a14
a15
4
a8
b8
a9
b15
S5
b10
a10
a17
b17
a11
b11
9
b18
END
a18
b12
b3
S2
b4
a4
a5
b5
S6
a2
a3
a16
b1
b2
b9
b16
S9
a1
S1
b14
S8
a7
b13
3
a12
MIDDLE
S3
b6
START
a6
S1-S9 :
Thermocouples
a1-a18: BioIndicators
(Bacillus
Subtilis)
b1-b18:
Endotoxin
challenged
containers.
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PART-2
STEAM STERILIZATION
VALIDATION
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STEAM STERILIZATION
Various types of steam sterilzers are
commercially available;
Saturated Steam
Water Immersion
Water Cascade System
Air-Steam Mixtures
Gravity Air Displacement (unpacked materials
sterilization)
Vacuum air Displacement (Packed materials)
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STEAM STERILIZATION
Microbiological aspects of Steam Sterilization
and Dry Heat Sterilization are basically the
same;
-D Value is determined in the same way
-Z Value = 10 0C
Tb = 121 0C
Lethality (F0 ) can be calculated in the same
way.
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Temperature
(0C)
100
103
106
109
112
115
118
121
121
121
118
115
112
109
106
103
100
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Facility layout.
Utility requirements and specifications.
Required capacity of the sterilizer.
Type of materials to be sterilized
(Liquids, wrapped ,hollow or porous materials)
Requirement for Gravity and/or Prevacuum
cycles.
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Temperature Distribution:
External monitoring and recording instruments
shall be calibrated before and after the OQ/PQ
studies (3 point calibration, 0.5 0C tolerance).
Uniformity of the temperature distribution in
case of Min&Max. loading should be verified by
using Thermocouples with 3 replicates.
T/C placement shall be documented on a
diagram.
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Temperature Distribution:
At least one T/C shall be placed located in the
steam exhaust line or adjacent to the
equipment temperature controller.
Min. 10-12 T/C s shall be used and they
should not be inserted in the load. Data should
be recorded during the whole cycle at 1 min.
intervals.
Location of the cold spot should be
determined and documented.
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STERILIZATION VALIDATION-GENERAL
CHANGE CONTROL AND REVALIDATION
Any changes to the sterilization equipment and/or
related utilities should be evaluated by a Change
Control Procedure.
Typical Changes Requiring Revalidation
Any changes in operating cycle (i.e:temperature ,
time, belt speed, chamber pressure)
Change in load configuration.
Change in sterilized materials.
Major maintenance work on critical
instruments/elements or utilities.
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..The END..
Thanks For Your Attention,
Any Questions Please?
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