1.0 IDENTIFICATION OF CRITICAL CONTROL MONITORING PARAMETER
Before
proceeding for Validation of Autoclave cum bung processor, following parameters
To be
Checked –
· Biological Indicator spore concentration must
be 106.
· Used media must be tested for its growth
promoting test.
·
Calibration of Pressure Gauge
· Calibration of Temperature Gauge
· Calibration of PT -100 Sensors
2.0 VALIDATION PROCEDURE
Validation
of an Autoclave will be considered qualified for consistent and reliable performance (validated) on successful completion of the following –
· Vacuum Leak Test
· Bowie-Dick Test
· Heat Distribution Study
· Heat Penetration Study
·
Biological Challenge Test
·
Bung Processing
· Flip off Seal
To
qualify these tests, the equipment should fulfill the acceptance criteria
described in the individual test procedures.
3.0 GENERAL CONSIDERATION/PREREQUISITE
A. Approved standard operating procedure of the equipment shall be availableB. Approved analytical methods for testing the samples collected during the processing
C. The impact analysis of the equipments shall be recorded in the summary sheet.
D. The installation and operational qualification of the equipment shall be successfully completed before the execution of the performance qualification.
E. All the deficiencies and discrepancies related to the equipment which affect the product quality and corrective action taken shall be recorded in the appropriate section of the protocol.
F. The analytical test results and other reports related with the equipment shall be attached with the performance qualification of the equipment and finally verified.
3.1 CALIBRATION DETAIL OF TEMPERATURE SENSORS
Equipment
Name
|
Equipment I.D
|
Data of calibration
|
Due
Date
|
Date of Pre
calibration
performed
|
Date of
Post
calibration
performed
|
Done by
|
Checked
by
|
3.2 CALIBRATION OF TEMPERATURE SENSORS:
• Pre & Post Calibration of Temperature
Sensors.
• Pre & post calibration shall be carried
out before starting and after completion of heat distribution cycles as well as
heat penetration cycles.
PRE & POST CALIBRATION OF TEMPERATURE SENSORS:
• PREPRATION OF ICE BATH:
Prepare a container with crushed ice and add enough purified
water to ensure a proper
Slush allow the temperature to stabilize ensure to add
sufficient crushed ice to maintain the equilibrium state of ice and water.
• PROCEDURE :
Temperature
sensors which are used for qualification study shall be calibrated in ice bath at
approximately 0oc and in high temperature reference block at
50,100,121,150oc prior to its usage in qualification.
Record
the temperature of all the sensors while putting them in ice bath after one minute
of temperature stabilization. Put
the individual sensor to the slot of high temperature reference block which is
stabilized at required temperature. Record
the temperature for five minute by data logger and attach the report with print
outs with reports.
ACCEPTANCE CRITERIA:
No temperature sensors vary by 1oc in ice bath from the
mean of temperatures shown by the calibrated thermometer during the data
logging period.
No temperature sensors vary by 1oc in high temperature
reference block from the mean of temperatures Shown by the calibrated
thermometer during the data logging period.
4.0 RE-VALIDATION CRITERIA:
Replacement
of major component / instrument.
Major
modification in the existing equipment / utility.
During
monitoring if system is found to be malfunctioning.
Shifting of the equipment/Instrument from one location to another
5.0 SYSTEM DESCRIPTION:
Autoclave cum Bung processor is designed to sterilize the
components which are used in the Aseptic area while processing the sterile
products. Autoclave cum Bung processor having the chamber size 900×900×1500mm
and is made up of SS 316L provided with 3 validation ports having 7 Nos of
probes in Each validation port. Argon welding is done in joints of piping. Vacuum
Break filter 5 inches long Having 0.2 micron porosity is provided Bung & flip
off carriage made of SS 316L, 2 Nos. trolley made of SS 304L for keeping the components in autoclave cum bung processor provided during
sterilization. All the system is controlled through PLC.
6.0 PERFORMANCE QUALIFICATION:
The Autoclave Cum Bung Processor will be qualified after
validating (As per the methods outlined in this Protocol) the equipment for
desired performance and its ability to sterilize different components and/or
loads at the set parameters & set loading patterns, repeatedly &
consistently.
The Autoclave Cum Bung Processor will be considered qualified for
consistent and reliable performance (Validated) on successful completion of the
following tests.
· Vacuum Leak test (3 Trials with probe)
· Bowie –Dick Test for Steam penetration (3
trials on 3 different days)
· Steam qualification tests (3 trials on 3
different days)
· Empty Chamber Heat Distribution studies (3
trials) with temperature mapping probes at different locations of the
sterilizer chamber.
· Loaded Chamber heat penetration studies (3
trials) for each sterilization load of fixed loading pattern, with temperature
mapping probes inside the innermost possible layer of the load subjected for
sterilization.
· Bio-Challenge studies using Bacillus
stearothemophilus spore strips (containing 106 or more spores
per strip) during the heat distribution & heat penetration studies.
· Estimation of the FO value
achieved during the sterilization hold period at each temperature-mapping
probe.
· Quality of the steam condensate collected
from the sampling point provided in the sterilizer chamber condensate
drain line for all loads.
· Vacuum break filter integrity
testing.
To qualify these tests the equipment should fulfill the
acceptance criteria described in the individual test procedures.
6.1 VACUUM LEAK TEST
• Objective:
To
verify the leakage in sterilization chamber during Vacuum Hold when the
sterilization chamber is empty.
• Procedure:
· Operate the equipment as per SOP .
· Set the parameters in PLC.
PROCESS PARAMETERS:
PRE
VACCUM
:
DELAY
BEFORE HOLD :
VACCUM
HOLD TIME :
CHAMBER
PRESSURE :
· Record the vacuum Hold start pressure and
vacuum Hold end pressure & calculate the actual leakage by subtracting the
former from later.
Three
consecutive cycles shall be carried out as per above parameters and procedure.
• ACCEPTANCE CRITERIA
Vacuum leak rate should not be more than 0.013 BAR / 10
minute.
• RESULTS:
Record the Observations in Annexure # 1
6.2 STEAM QUALITY TESTS
A continuous supply of saturated steam is required for steam sterilization. Too high a level of non-condensable gases will prevent the
attainment of sterilizing conditions; too little moisture carried in suspension
may allow the steam to become superheated during expansion into the chamber,
while excess moisture may cause damp loads.
For all these tests, the steam should be sampled from the steam
service pipe. The measurements are taken during a period of maximum steam
demand, when steam is first admitted to the sterilizer chamber.
Presence of non-condensable gases more than the desired level
might get accumulated within the sterilization chamber in the form of air
pockets, which might affect steam penetration this leading to probable
non-sterile unit.
NOTE: Silicone rubber tubing is porous to steam and should not be
used to carry steam in these tests.
6.2.1 Non-Condensable Gas Test
Objective
Objective of this test is to ensure that,
· The pure steam supply of the High Pressure
High Vacuum Steam Sterilizer does not contain non-condensable gases more than
the desire level (NMT 3.5%) when measured on-line during the standard
sterilization cycle.
Procedure
This test is used to demonstrate that the level of non-condensable
gases in the steam will not prevent the attainment of sterilization conditions
in any part of the load. The method described should be regarded not as
measuring the exact level of non-condensable gas, but a method by which the
provision of acceptable steam quality can be demonstrated. The apparatus is shown and described in Figure No.1 (all sizes are
nominal). Connect the needle valve to the steam service pipe as shown in Figure
No.1. Assemble the apparatus so that condensate will drain freely from
the long rubber tube into the sampling pipe. If the tube is too short, copper
or stainless steel tubing may also be used. Fill the container with cold water until it overflows. Fill the
burette and funnel with cold water, invert them and place them in the
container. Draw out any air that has collected in the burette. With the steam
sampling pipe out of the container, open the needle valve and allow steam to
purge the air form the pipe. Place the pipe in the container, locate the end
within the funnel, and add more cold water until it flows through the overflow
pipe. Place the empty measuring cylinder under the container overflow. Adjust the needle valve to allow a continuous sample of steam into
the funnel sufficient to cause a small amount of “Steam Hammer” to be heard.
Ensure that all the steam is discharged into the funnel and does not bubble out
into the container. Note the setting of the needle valve. Close the valve.
Ensure that the container is topped up with cold water and that
the measuring cylinder is empty. Draw out any air present in the burette. Ensure that the sterilizer chamber is empty except for the usual
chamber furniture. Select and start the operating cycle. When the steam supply to the chamber first opens, open the needle
valve to the previously noted setting, allowing a continuous sample of steam
into the funnel sufficient to cause a small amount of steam hammer to be heard. Allow the steam sample to condense in the funnel. Any
non-condensable gases will rise to the top of the burette. Overspill formed by
the condensate and the water displaced by the gases will collect in the
measuring cylinder.
When the temperature of the water in the container reaches 70-75oc close the needle valve. Note the volume of gas collected in the burette (Vb)
and the volume of water collected in the measuring cylinder (Vc).
Calculate the fraction of non-condensable gases as a percentage as
follows.
When the temperature of the water in the container reaches 70-75oc close the needle valve. Note the volume of gas collected in the burette (Vb)
and the volume of water collected in the measuring cylinder (Vc).
Calculate the fraction of non-condensable gases as a percentage as
follows.
Fraction of non-condensable gases = 100 x (Vb/Vc).
The test should be considered satisfactory if the fraction of
non-condensable gases does not exceed 3.5 %.
The test should be done two more times to check consistency. If
the results of the three tests differ significantly, then the cause should be
investigated before proceeding further.
Figure No.1
Acceptance Criteria:
The measured non-condensable gases in the pure steam should not
cross 3.5%
Observations & Results
Record the observations and results in formats enclosed as
Attachement-2, Annexure-1
6.2.2 Super Heat
Objective
This test is used to demonstrate that the amount of moisture in
suspension with steam from the service supply is sufficient to prevent the
steam from becoming superheated during expansion into the chamber.
The method described here uses a low-volume sample, continuously
taken from the center of the steam service pipe. The level of superheat
determined by this method cannot be regarded as indicative of the true dryness
of the steam in the pipe since condensate flowing along the inner surface is
not collected. However, devices designed to separate free condensate are
incorporated into the steam delivery system to the chamber and therefore the
level determined by this method is representative of steam conditions likely to
prevail within the chamber during the plateau period.
Procedure
This test should normally follow a satisfactory test for non-condensable
gases.
This test, and the subsequent dryness value test, requires a pitot
tube as shown in figure No.2. The rest of the apparatus is shown and described
in figure No.3. All sizes are nominal. Fit the Pitot tube concentrically within the steam service pipe as
shown in figure No.3. Fit the sensor entry gland to the steam service pipe. Insert one
of the sensors through the gland and position the axis of the pipe. Insert the second sensor through the gland in the expansion tube
and position it on the axis of the pipe. Wrap lagging around the expansion
tube. Push the tube on to the pitot.
Figure No.2.
Ensure that the sterilizer chamber is empty except for the usual
chamber furniture. Select and start the operating cycle.
From the measured temperatures, note the temperature in the steam
service pipe (for use in the dryness test) and in the expansion tube (Te) when
the steam supply to the chamber first opens. Calculate the superheat in oC
from the following equation:
Superheat = Te - To
Where:
To is the boiling point of water at local atmospheric pressure.
The test should be considered satisfactory if the superheat
measured in the expansion tube does not exceed 25oC.
Record the observations and results in formats enclosed as
Attachement-2, Annexure-2.
6.2.3 DRYNESS TEST
The accurate measurement of the percentage of moisture content in
the steam is difficult, and the traditional methods where constant steam flow
is required are not suitable for sterilizers. This test should be regarded not
as measuring the true content of moisture in the steam, but as a method by
which the provision of acceptable steam quality can be demonstrated.
6.2.4 Objective
This test is used to demonstrate that the dryness value is not
less than 0.90 (if metal loads be processed, the dryness value should not be
less than 0.95); throughout the operating cycle, the temperature measured in
the steam service pipe is within 30C of that measured during
the superheat test.
6.2.5 Procedure
The test is conveniently carried out immediately after the
superheat test.
This test requires a pitot tube as shown in Figure No.2. The
apparatus is shown and described in figure No.4. All sizes are nominal. A
laboratory balance is also required, capable of weighing a load up to 2 kg with
an accuracy of 0.1g or better. If it is not already fitted, fit the Pitot tube concentrically within
the steam service pipe as shown in figure No.3. If it is not already fitted, fit the sensor entry gland to the
steam service pipe. Insert a temperature sensor through the gland and position
it on the axis of the pipe.
Connect the rubber tube to the longer of the pipes in the stopper,
place the stopper in the neck of the vacuum flask, weigh the whole assembly and
note the mass (M1). Remove the stopper and tube assembly and pour 650 +50 ml of
cold water (below 27oC) in to the flask. Replace the stopper and
tube assembly, weigh the flask and record the mass (M2). Support the flask close to the pitot, and ensure that the rubber
tube and flask are protected from excess heat and draughts, do not connect it
to the pitot tube yet.
Introduce the second temperature sensor through the shorter of the
two pipes in the stopper and into the water in the flask. Note the temperature
of the water in the flask (To). Ensure that the sterilizer chamber is empty except for the usual
chamber furniture. Select and start the operating cycle.
When the steam supply to the chamber first opens, connect the
rubber tube to the pitot discharge and wrap lagging around it. Arrange the
rubber tube to permit condensate to drain freely into the flask. Not the
temperature in the steam service pipe (TS).
When the temperature of the water in the flask is approximately 80oC,
disconnect the rubber tube from the pitot, agitate the flask so that the
contents are thoroughly mixed, and note the temperature of the water (T1).
Weigh the flask and stopper assembly and note the mass (M3).
The initial mass of water in the flask is given by Mw = M2 – M1
The mass of condensate collected is given by MC = M3 -
M2.
Calculate the dryness value of the steam from the following
equation:
D= (T1-T0) (4.18MW + 0.24) / LMC - 4.18
(TS-T1) / L
Where:
T0 = Initial temperature of the water in the flask (oC);
T1 = Final temperature of the water and condensate in the flask (oC);
TS = Average temperature of the steam delivered to the sterilizer
(oC);
MW = Initial mass of water in the flask (Kg);
MC= Mass of condensate collected (Kg);
L= latent heat of dry saturated steam at temperature TS (kJ Kg-1).
6.2.6 Acceptance Criteria
The test should be considered satisfactory if the following
requirements are met.
A) The dryness value is not less than 0.90 (if metal loads are to
be processed, the dryness value should not be less than 0.95);
B) Throughout the operating cycle, the temperature measured in the
steam service pipe is within 3oC of that measured during the
superheat test.
6.2.7 Observation and results
Record the observations and results in formats enclosed as Attachement-2,
Annexure-3 See next page for fig no.4.
6.3 BOWIE-DICK TEST
• Objective:
To
ensure that the vacuum pulses applied before the sterilization hold period are
sufficient to remove the entrapped air or non condensable gases so as to
facilitate the event and rapid steam distribution into all parts of load and
maintaining these conditions for specified temperature holding time.
6.3.1 PROCEDURE
· Operate the equipment as per SOP.
· Set the following parameters in PLC.
PRE
VACUUM
PRE
PRESSURE
STERILIZATION
HOLD TEMPERATURE
STERILIZATION
HOLD TIME
· Place one Bowie dick test kit pack
in the center (Near drain) of the sterilization chamber supported
approximately 100 to 200 mm above the sterilization chamber base.
· The printouts taken
during Bowie dick test cycle & bowie dick indicator
shall be attached as per exhibit.
· Compile the observation and take three
consecutive cycles.
Acceptance
Criteria
The Bowie Dick test indicator shows the uniform color change. No
change, no uniform change and / or Air entrapment (bubble) spot on the test
pack indicate inadequate air removal from the sterilization base chamber.
Results
6.4 HEAT DISTRIBUTION STUDY (EMPTY CHAMBER)
• Objective:
Objective
of this test is to verify that the temperature uniformity throughout the
chamber and to Locate the cold spot in empty Chamber.
· The sterilizer is capable of attaining the temperature
of 121.0oc during the sterilization
Hold
period.
· The profile point having the lowest
temperature or slowest to heat is designated as the cold spot.
Procedure:
Insert
16 nos. of temperature sensors inside the chamber through the validation port of
sterilizer .seal the port with silicone sealant to ensure that no steam leakage
during operation of sterilizer.
Fix
the probe at the location at the location in the sterilizer so that sensors do
not touch the metallic surface of the chamber. Connect the temperature sensors
to the data logger which can scan and print the actual temperature at different
locations. Set the following parameters & operate the autoclave cum bung
processor and also start the data logger to record the actual temperature.
SET
PARAMETERS :
Sterilization
Hold temp = 121.0oc
Sterilization
Hold time = 30 min
Chamber
high pressure = 1.5 BAR
· After completion of sterilization collect
thermograph from the multipoint temperature recorder of the autoclave cum bung
processor.
· Download the data from data logger to the
computer for data analysis & graph prep.
· Perform the three consecutive cycles to
demonstrate the consistency of sterilizer.
• ACCEPTANCE CRITERIA:
1. Throughout
the cycle time, all temperature measured in the chamber do not fluctuate by
more than 20C.
2. Throughout
the cycle time, all temperatures measured in the chamber do not differ from
each other by more than 20C.
3. The
interval of time between the attainment of the sterilization temperature in the
hottest and coldest part of the chamber does not exceed 60 seconds.
• Results: Record the observations.
LOCATION
OF TEMPERATURE SENSORS INSIDE THE CHAMBER
SENSOR NO.
|
LOCATION OF SENSORS IN THE CHAMBER
|
S1
|
In the drain of
autoclave chamber
|
S2
|
Lower left
front corner of the non sterile side
|
S3
|
Upper left
front corner of non sterile side
|
S4
|
Upper right
front corner of non sterile side
|
S5
|
Lower left
front corner of non sterile side
|
S6
|
Middle left
side of the chamber
|
S7
|
Middle right
side of the chamber`
|
S8
|
Middle front
non sterile side of the chamber
|
S9
|
Middle back
sterile side of the chamber
|
S10
|
Lower left
sterile side of the chamber
|
S11
|
Upper left
sterile side of the chamber
|
S12
|
Upper right
sterile side of the chamber
|
S13
|
Lower right
sterile side of the chamber
|
S14
|
Middle of upper
most surface of the chamber
|
S15
|
Middle of lower
surface of the chamber
|
6.5 HEAT PENETRATION STUDY
• Objective
Objective
of this test is to ensure that, the steam is sufficiently penetrating into the
innermost portion of the load subjected for sterilization to achieve desired
temperature i.e. 121.40C during the sterilization cycle.
Heat
penetration studies shall be carried out with the following different loads:
a. Maximum
garment load
b. Minimum
garment load
c. Accessory
load (filling machine parts load)
d. Bung
load
e. Flip
off seal
Temperature
probe and biological indicator placement in the maximum garment
load
Load
details –
Sterile
garments - 24 pieces
Booties - 48 Nos.
Head
Gear -
24Nos.
Gloves
(7 inches) - 24 pairs
LOAD
CONFIGURATION:
One
Perforated box (C1) containing 8 pair of garments placed on lower front of
trolley (non sterile door side).
One
Perforated box (C2) containing 8 pair of garments placed on lower back side of
trolley (sterile door side)
One
Perforated box (C3) containing 8 pair of garments placed on upper front of
trolley (non sterile side)
One
perforated box (C4) containing 24 pair of gloves wrapped in parchment paper &
04 pair of garments placed on upper back side of trolley (sterile door side)
Temperature
Sensors with Chemical & Biological Indicator Placed In Maximum Garment Load
Sensor no.
|
Location of sensors in the chamber
|
S1
|
In the drain of
autoclave chamber
|
S2
|
Center of the
garment pack located at the lower surface in C1
|
S3
|
Center of
garment pack located in the middle of C1
|
S4
|
Center of the
garment pack located at the upper surface in C1
|
S5
|
Center of the
garment pack located at the lower surface in C2
|
S6
|
Center of the
garment pack located in the middle of C2
|
S7
|
Center of the
garment pack located at the lower surface in C2
|
S8
|
Center of the
garment pack located at upper surface in C2
|
S9
|
Center of the
garment pack located at lower surface in C3
|
S10
|
Center of the
garment pack located in middle of C3
|
S11
|
Center of the
garment pack located in middle of C3
|
S12
|
Center of the
garment pack located at upper surface in C3
|
S13
|
Center of the
garment pack located at lower surface of C4
|
S14
|
Center of the
garment pack located at upper surface in C4
|
S15
|
Center of
gloves pack in C4
|
Temperature
probe and biological indicator placement in the minimum garment load
Load
Details
Sterile
garments – 6 pairs
Gloves
(6 inches) – 8 pairs
Load
configuration:
One
perforated box containing 6 pairs of garments & 8 pair gloves wrapped in
parchment paper placed on lower middle of chamber.
TEMPERATURE
SENSORS WITH CHEMICAL & BIOLOGICAL INDICATOR PLACEMENT IN
PARTIAL GARMENT LOAD
SENSOR NO.
|
LOCATION IN THE CHAMBER
|
S1
|
In the drain of
autoclave chamber
|
S2
|
Center of the
garment pack located at the lower surface in C1
|
S3
|
Center of the
garment pack located in the middle of C1
|
S4
|
Center of the
garment pack located at the upper surface in C1
|
S5
|
Center of glove
pack located at upper surface in C1
|
Temperature
probe and biological indicator placement in the accessory load (Filling machine
parts)
• Load details
· Powder Hopper- 2 Nos.
· Piston
Wheel- (2 Nos.)
·
Piston- (32 Nos.)
· Piston Tips- (36 Nos.)
· Spoon-(2 Nos.)
· Scoop-(2 Nos.)
· Tubes (4 Nos.)
· Bung Boxes- (2 Nos.)
· Forceps- (4 Nos.)
· Sterile Garments – 12 pieces
· Booties – 12 pairs
· Gloves (7 inches) – 24 pairs
• Load configuration:
· 12 sterile garments, 12 Booties & 24
Pairs Gloves (6 inches) in SS Perforated Container placed on Top.(Non Sterile
Side)
· One Perforated box containing 2 Piston Wheel,
32 Nos. Piston and 36 Nos. Piston Tips., Placed on Bottom.(Sterile Side)
· One Perforated box containing 4 Nos. Tubes, 4 Nos. forceps, 2 Nos. Spoons, 2 Nos. Scoops, Placed on
Top.(Sterile Side)
· Powder Hopper 2 Nos. & 2 Nos. Bung Boxes
Placed at Bottom (Non Sterile Side).
6.5.1 PROCEDURE
Conduct
the study with loaded chamber cycles with temperature probes and Biological
Indicators. Transfer the load to sterilizer and connect the 15 probes &
Biological Indicators as per the locations. Connect the outputs of all the
probes to the temperature data logger and close the door of sterilizer. Switch ON the MAINS of the control panel and
set the parameters –
Temperature
: 121.4°C
Chamber
Pressure High : 1.5
BAR
Hold
time : 30 minutes
Simultaneously
Insert new chart in chart recorder provided on the control panel of an autoclave
and adjust the start time and temperature of the instrument. Now start the
cycle as per SOP for Operating Instruction. After attaining temperature 121.40C,
record the chamber temperature and pressure for every minute. Simultaneously
start the recording with data logger and take print outs
At
the end of the cycle Switch OFF the cycle. When pressure becomes -0.04 BAR,
open the door with the help of safety gloves. Remove the load. Repeat
the same cycle three times and compile the data.
6.5.2 Acceptance Criteria:
Throughout
the cycle time all temperature measured in the chamber do not fluctuate by more
than 20C.
Throughout
the cycle time, all temperatures measured in the chamber do not differ from
each other by more than 20C.
The
calculated minimum F0 value should be more than biological F0 value
for the Biological indicator.
Each
autoclaved Biological indicator gives the ‘-ve’ test during the incubation.
Results:
Record
the Observations.
6.6 Biological Challenge Test
Objective
To
demonstrate the degree of Process Lethality provided by the Sterilization
Cycle.
Test
Requirement
Spores
of Geobacillus Stearothermophilus
Procedure
After
determining the worst case items and worst locations i.e. cold spots, challenge
these items/locations with biological indicator (spore Geobacillus Stearothermophilus)
Carry
out the Microbial Challenge Studies concurrently with Loaded Chamber Heat penetration
studies.
Previously
population validated biological ampoule indicators of specified 106 or more
spores of Geobacillus Stearothermophilus
should be issued for validation as per procedure.
Place
the biological indicators along with the probes at the same locations,
within each load type of the specified loading configuration and pattern,
as in the Loaded Chamber Heat penetration studies. Retain 2 biological
indicators as positive controls. Operate the Autoclave cum Bung Processor as
per SOP.
Switch
ON the Mains of the control panel and set the parameters –
Temperature
: 121.4°C
Chamber
Pressure High : 1.5 BAR
Hold
time
: 30 minutes
Simultaneously
insert new chart recorder provided on the control panel of an Autoclave and
adjust the start time and temperature of the instrument.
Now
start the cycle.
After
attaining temperature 121.4°C, record the chamber temperature and pressure for every
60 seconds.
Simultaneously
start the recording with data logger and take printouts. At the end of the
cycle Switch OFF the cycle. When pressure becomes -0.04 BAR,
open the door of an Autoclave with the help of safety gloves. After incubation observe
the indicator for growth. (+ve when purple color change to yellow color, -ve
when purple color remain as such) Conduct the test until a
cycle time results in three consecutive runs where the biological indicators
show on growth.
• Acceptance Criteria
1. Visually observe the ampoules,
test +ve when purple color change to yellow color, test -ve when
purple color remain as such).
2.
If no
evidence of growth observed in any of the inoculated tube and growth observed
in positive control tube, the test meets the criteria to achieve the Sterility Assurance Level (SAL) 10-6.
6.7 Bung Processing
Objective:
Objective
of this test is to ensure that, the bung washing is proper and bung load
subjected for sterilization to achieve desired temperature i.e. 121.4°C during
the sterilization cycle.
Load
details
· Maximum Bungs cycle (Approx 80000)
· Minimum Bungs cycle (Approx 5000)
Load
configuration
· Bungs in Bung Carriage (Approx 80000)
6.7.1 Procedure
• Load the bungs in Bungs Carriage cassettes.
Approximately 8000 Bung Load in each cassette.
• Place one biological indicator in each
cassette and transfer the load to Autoclave cum Bung processor.
• Switch ON the MAINS of the control panel and
run cycle as per HPHV Process Parameters.
Simultaneously
Insert new chart in chart recorder provided on the control panel of an autoclave
and adjust the start time and temperature of the instrument after attaining
temperature 121.4°C, record the chamber temperature and pressure for every
minute. At
the end of washing cycle send WFI rinse sample to QC for Analysis of
Non Ionic Detergent Content. At the end of the cycle switch OFF the cycle.
When
pressure becomes -0.04 BAR, open the door with the help of safety gloves. Remove
the load. Repeat the same cycle three times and compile the data.
Acceptance
criteria:
The
calculated minimum F0 value should be more than biological F0 value
for the Biological indicator. Each autoclaved biological indicator gives the
‘-ve’ test during the incubation. WFI rinse sample should be free from non
ionic Detergent.
Results:
Record
the Observations
7.0 F0 Calculation
The actual observations obtained during the heat penetration
studies at different temperature sensing locations were compiled in the table
and the observed temperature were subjected for calculation of F0 values at
that particular location. The calculations are done by using the following
formula and the lethality factor computed (during the sterilization period) are
given in the following table.
F0=dt
∑ 10(T-121)/Z __________ (a)
F0=dt
∑ (Sum of lethality factors)
Where,
Dt =
The time interval between successive temperature measurements (1 min).
T
= The observed temperature at that particular time (as per the actual
temperatures recoded
Z
= The change in the heat resistance of Bacillus
stearothermophilus spores as temperature is Changed (10°C)
The calculated minimum F0 value (by equation a)
should be more than the biological F0 value for the biological
indicator strip exposed for the bio-challenge study. The biological F0 value for biological indicator strip exposed during the sterilization can be
calculated as follows.
F0 = D121 (log A – log B) ……… (b)
Where,
D121
|
D value of the
biological indicator at 1210C
|
|
A
|
Biological
indicator concentration or spore population
|
|
B
|
Desired level
of sterility
|
8.0 INTERPRETATION OF RESULTS
All test results shall be compiled and evaluated. Compliance of
all the results with empty chamber and loaded chamber with different loads to
the acceptance criteria shall establish that the sterilization practices,
control and facility are suitable for various required loads.
9.0 REVALIDATION FREQUENCY
Schedule Revalidation
Twice
in a
year.
Unscheduled Revalidation
Unscheduled revalidation shall be carried out in following
situation:
When major changes in the load is taken place.
After major maintenance of existing equipment.
10.0 DOCUMENTATION
Results
and reports shall be compiled in a binder. Binder shall contain the following
sequentially:
Summary Report
Test Reports
11.0 CONCLUSION
Based
on the rigorous studies performed during the execution of the qualification
protocol and as per the results obtained, it is concluded that the Autoclave
cum Bung Processor is qualified / not Qualified for consistent
results.
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