PROTOCOL FOR QUALIFICATION O D H S F A E T N .:……………. A M L · 2020. 12. 21. · 5.3.2.2 Validation Port The unit is provided with a 1 Validation port for the insertion

PROTOCOL No. PERFORMANCE QUALIFICATION

PROTOCOL

FOR

DRY HEAT STERILIZER

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EFFECTIVE DATE

PAGE No. Page 1 of 40

PHARMA SCHOLARS MICROBIOLOGY DEPARTMENT

PROTOCOL

FOR

QUALIFICATION

OF

DRY HEAT STERILIZER, FULLY AUTOMATIC

EQUIPMENT TAG NO.:…………….

AREA: MICROBIOLOGY LAB

Document No.

Supersedes

Effective Date

No. Of Pages 41


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PROTOCOL CONTENTS

S. No. Section Title Page No.

1.0 Protocol Approval ......................................................................... 3

2.0 Overview ......................................................................................... 4

2.1 Objective .............................................................................. 4

2.2 Purpose and Scope……………………..………………….4

2.3 Responsibility ...................................................................... 4

2.4 Qualification Team .............................................................. 4

3.0 Training Record ............................................................................ 5

4.0 Qualification Requirements .......................................................... 6

5.0 System / Equipment Description .................................................. 7

5.1 System / Equipment details ................................................ 7 5.2 System/Equipment Identification/Description…………...7

5.2 Generic Design .................................................................... 7

5.3 Safety features ................................................................... 10

6.0 Performance Qualification Procedure ....................................... 12

6.1 Air Velocity Test………………………………………….12

6.2 HEPA Filter Integrity Testing…………………………...13

6.3 Air Flow Pattern Test…………………………………….14

6.4 Heat Distribution Study………………………………….15

6.5 Heat Penetration Study…………………………………..15

6.6 Endotoxin Challenge Study……………………………...18

6.7 Non-viable Air Borne Particle Count.…………………..19

7.0 Acceptance Criteria ..................................................................... 20

8.0 Qualification Report .................................................................... 21

9.0 Approval of Qualification Report .............................................. 21

10.0 Observed Deviation ..................................................................... 22

11.0 List of Exhibits/Annexure ........................................................... 23

11.1 List of Exhibits .................................................................. 23

11.2 List of Annexure ................................................................ 23

12.0 Reference Documents ................................................................. 23


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1.0 Protocol Approval

This is a specific protocol for Qualification of Dry Heat Sterilizer (Fully Automatic)

which is installed, in the New Microbiology Lab.

Initial Approval

This protocol has been approved by the following:

Prepared By:

Name Designation Department Signature Date

Checked By:


Final Approval:



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2.0 Overview

2.1 Objective

To establish the methodology for the Qualification of Dry Heat Sterilizer (Fully Automatic).

2.2 Purpose and Scope

The purpose of this protocol is to establish Documentary evidence that Dry Heat Sterilizer

(Fully Automatic) has been qualified to perform Dry Heat Sterilization.

This Protocol is Applicable for the Qualification of the DRY Heat Sterilizer (Fully Automatic)

which is installed in the Autoclave and DHS loading room in Microbiology Lab.

2.3 Responsibility

• Protocol / Report Preparation: Microbiology Executive

• Execution of Qualification Activity: Microbiology Executive/ QA Executive

• Protocol / Report checking : Manager QC / Manager QA

• Approval of Protocol / Report: Quality Control Manager / QA Manager and above

2.4 Qualification Team

• Microbiology Executive / Manager

• Quality Control Manager/Head

• Quality Assurance Executive / Manager


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3.0 Training Record

3.1 Purpose

The purpose of the training is to familiarize the trainees with the overall strategy of

performance qualification of Dry Heat Sterilizer (Equipment No.:………).

3.2 Scope

This Training is applicable to the Dry Heat Sterilizer (Fully Automatic).

3.3 Topics

The following topics shall be covered during training:

• Overall strategy of qualification process.

• General precautions/guidelines to be followed during qualification.

• Training records shall be attached with the report as Annexure – 01.


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4.0 Qualification Requirements

Following instruments shall be calibrated at the time of the Re-Qualification.

S.No. Instrument Name Instrument

Code/Sr. No.

Calibration

Certificate No.

Calibration

Due on

1. Inbuilt RTD sensors








9. Temperature indicator

controller of panel

10. External Data Logger

11. RTD Sensor of Data Logger
















27. Aerosol Photometer

28. Non Viable particle counter

Calibration Certificate shall be attached as Annexure-06.


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5.0 System / Equipment Description

5.1 System / Equipment details

The Dry Heat Sterilizer (Fully Automatic) Equipment ID No.:................ shall be used for

the Depyrogenation of BET tubes and Sterilization of Other glassware in Microbiology Lab.

5.2 System /Equipment Identification/Description

Component Specifications

Name of equipment Dry heat Sterilizer (Fully Automatic)

Equipment ID No -----

Serial Number -----

Name of the Manufacturer Fabwell Engineers

Dimensions 600 (W) X 900 (H) X 900 (D) mm

Working Temperature Up to 2800C

Chamber Volume 0.486m3

Doors Hinged

Details of Purchase Order No. ------

Equipment Location New Microbiology laboratory

5.3 Generic Design

5.3.1 Working Principle

The Dry Heat Sterilizer (Fully Automatic) can be efficiently used to perform Dry Heat

Sterilization and Depyrogenation processes: It is base on following two principles.

5.3.1.1 Heat Conduction:

Conduction is accomplished either 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 conductor of heat.


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5.3.1.2 Radiant Heating Process:

Radiant heating is the process in 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.

5.3.2 Brief Machine Description

5.3.2.1 Chamber & Outer Cover

The outer body of the DHS is made of Stainless steel and inside chamber is also made out of

stainless steel sheet. The inside of the DHS is tapered to get uniform air circulation.

Portion between the two walls is having a mild steel framework. Top sheet is of Stainless

Steel for mounting re-circulation fan motor, Dampers and also for junction box.

5.3.2.2 Validation Port

The unit is provided with a 1 Validation port for the insertion of flexible RTD, PT-100

sensors. The Port end connections is Flange type with gasket for ease of insertion

5.3.2.3 Insulation

The chamber is insulated with fibre-glass. The insulation is held in place by a stainless steel

cover. This ensures less heat transfer to outside & also safety of handling.

5.3.2.4 Door

The equipment is having Two Stainless Steel Doors made. S.S. doors will also be insulated

with fibre glass with and covered with S.S. on the outer side. Doors are rugged and hinged to

external wall of Sterilizer frame-work for ease of opening.

5.3.2.5 Door Components

Other Door components such as Radial Locking Arms, Hinge Plate etc. are also of Stainless

Steel.


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5.3.2.6 Door Gaskets

The door gasket is of Silicone rubber (square cross section) fitted in the Groove provided on

the door.

5.3.2.7 Heating System

A heater of S.S. has been provided inside the chamber. Heaters have been provided in two

zones. Each Zone is controlled separately.

5.3.2.8 Air Circulation

The air inside the sterilizer has been provided Hot Air Circulation arrangement consists of a

blower fan and motor.

5.3.2.9 Blower Fan

Blower fan is made of S.S. 316 & is dynamically balanced. Suitable Silicon gasket has been

provided for motor and motor mounting plate.

5.3.2.10 Motor

It has been supplied with TEFC Motor of 3 HP, 1440 rpm, operable on 3 phase, 415 Volts,

50 cycles.

5.3.2.11 Filter Module

The DHS is provided with pressure module mounted on to the sterilizer which maintains

positive pressure inside to ensure that no outside air enters the sterilizer. It consists of

Pre-filter of 5 micron. & HEPA filter of 0.3 micron.

5.3.2.12 Exhaust Duct

The sterilizer is provided with exhaust duct with HEPA filter necessary Valves etc. to

exhaust hot air. The exhaust air can be taken out of the room by means of suitable duct

(clients supply).


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5.3.2.13 Exhaust damper

An Exhaust damper is provided at the top of the unit. The damper during the heat-up phase

opens to ensure complete removal of moisture and during the cooling phase.

5.4 Safety Feature Description

5.4.1 Alarms and Warnings

The DHS shall be equipped with several alarms and warnings which alert the operator (or the

User’s main control system) that system operation is outside of established parameters. Fatal

alarms switch the system into the shutdown sequence. Non-fatal warnings allow the system to

continue to run, but it will display “warning” messages and sound the alarm horn.

Alarms and warnings are separated into “fatal alarms” and “non-fatal warnings” based on the

Following criteria:

• Fatal alarms shut the system down when there is a reasonable risk of damage to the

system or of contamination of, or damage to, the user’s distribution loop or final product or a

reasonable risk of injury to personnel.

• Non-fatal warnings do not shut the system down, as there is little or no risk of damage

to the system or of contamination of, or damage to, the user’s distribution loop or final

product and little or no risk of injury to personnel.

5.4.1.1 Fatal Alarms

An alarm horn shall sound, and alarm message shall be displayed, and operation shall be

determinate.

When a fatal alarm occurs. Fatal alarms shall include:

• Door Open.

• Chamber high-temperature alarm

• Chamber low-temperature warning.

5.4.1.2 Non-Fatal Warnings

An alarm horn shall sound and a warning message shall be displayed when a non-fatal warning

occurs, but operation shall continue. Non-fatal warnings shall include:


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• Compressed air low-pressure warning

• Other Side Door Open warning

• Incorrect password entry warning

6.0 Qualification Procedure

The following procedure shall be used for the performance qualification of the Dry heat

Sterilizer Fully Automatic.

All the Calibration certificates shall be verified as per Exhibit-01 and the calibration

certificate shall be attached as Annexure-06. The Calibration and Temperature Verification

Checklist shall be as per Exhibit -E02

The following procedure shall be used for the Re-qualification of the Dry Heat Sterilization.

Air Velocity Test

HEPA Filter Integrity Test (PAO Test)

Air Flow Pattern Test

Heat Distribution Study

Heat Penetration Study

Endotoxin Challenge Study

Non Viable Air Borne Particle Count.

6.1 Air Velocity Test

6.1.1 Objective:

To demonstrate that the system is capable of delivering air velocities, as per the requirement, to

maintain continuous laminarity inside the Dry Heat Sterilizer.


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6.1.2 Procedure:

• Operate the Dry Heat Sterilizer as Per SOP, “Operation and Maintenance of Dry Heat

Sterilizer.

• Ensure that the blower is switch “ON” at least 20 minutes prior to the start of the

observations and keep the heaters bank “OFF”.

• Measure the air velocity at Distance of 6 inches from the filter face of each HEPA filter, at

5 locations (four corners and centre) with the Anemometer.

• Calculate the Average Velocity (V, in feet per minute) of each HEPA filter/ riser as:

Where, i = 1 to 5 and v = Velocity observed at each point.

Acceptance Criteria

The Average air velocity shall be within 90 ± 20% FPM.

Rationale for Acceptance criteria:

For Circulation of Temperature more air Flow required, hence more Air Required to Maintain

A Grade area.

The observations shall be recorded as per Exhibit – E10

6.2 HEPA Filter Integrity Testing (PAO Test)

1 2

5

3 4 4 3

2 1

5


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6.2.1 Objective

HEPA filter leak test shall be checked in order to find out that filters are not damaged and there

is no leakage from the filter media joints and frames etc.

PAO test shall be carried out as per test procedure stated below for leakage of HEPA filters. The

observations shall be recorded as per Exhibit–E08 & Test reports shall be attached as

Annexure-05.

6.2.2 Test Procedure:

• Filter integrity testing shall be checked using cold PAO (Poly alpha olfein) smoke generator

aerosol photometer

• Aerosol shall be introduced at the upstream side of HEPA filter, such that it gets the actual

concentration of 10 μg/L to 30 μg/L

• Actual upstream concentration shall be set as 100 %.

• The downstream of the filter shall be scanned with the probe at the speed of 10 FPM.

6.2.3 Acceptance Criteria:

Penetration shall not exceed 0.01% at 100 % upstream concentration.

6.3 Air Flow Pattern Test:

Objective:

To verify the unidirectional parallel airflow and non-turbulent up to working zone & movement

of air flow due the pressure gradient throughout the chamber maintaining the Class 100

environmental conditions.

6.3.1 Equipment / Instruments and Chemical Used:

Water based Smoke Generator


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6.3.2 Method applied:

• The temperature of chamber of the Dry Heat Sterilizer should not be more than 5°C above

the ambient temperature during the study.

• Ensure that blower of the Dry Heat Sterilizer is running and heater is OFF.

• Verify and record the path of smoke as per acceptance criteria.

• Above procedure can also be performed by using water fogger.

Acceptance criteria:

The smoke flow should be unidirectional from the HEPA filter face to return air riser.

The observations shall be recorded as per Exhibit – E11.

6.4 Heat Distribution Study

6.4.1 Objective

The Dry Heat Sterilizer when operated with Empty Chamber is capable of producing the

temperature profiles as per the temperature set points in the equipment and the temperature

distribution is uniform throughout the sterilization period when the chamber is empty.

6.4.2 Procedure

6.4.2.1 Heat distribution study of the Dry heat sterilizer shall be done at temperature 250 0C for not less

than 30 min using 16 flexible RTDs placed inside the chamber. Three runs of Empty cycle shall

be run at 250 0C for 30 min.

6.4.2.2 None of the RTD shall touch the body of the chamber of the sterilizer while doing RQ study.

6.4.2.3 Drawings showing location of the flexible RTDs in the chamber for heat distribution study shall

be as per Annexure-02.

6.4.2.4 Detail of each cycle run shall be recorded as per Exhibit-E03 and print outs shall be attached as

Annexure -07.

6.4.3 Acceptance criteria:

There should be uniform distribution of temperature NLT 250°C at each probe in sterilizer

chamber during sterilization hold period. Upper targeted limit for temperature will be 270°C.


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Based on validation study results for upper limit will be finalized in report.

6.5 Heat Penetration Study

6.5.1 Objective

Purpose of heat penetration studies is to check the penetration of heat into the loaded articles

when the Dry Heat Sterilizer when operated with loaded chamber is capable of producing the

temperature profiles as per the temperature set points in the equipment and the temperature

distribution is uniform throughout the sterilization period.

6.5.2 Procedure

6.5.2.1 Heat penetration study shall be done using not less than 16 flexible RTDs with the help of data

logger at 250 0C for not less than 30 min.

6.5.2.2 Three runs of sterilization cycle shall be run with loaded chamber to perform the heat

Penetration study.

Three runs of sterilization cycle with Minimum Load.

Three runs of sterilization cycle with Maximum Load.

6.5.2.3 Each probe shall be inserted inside the articles to be sterilized and one endotoxin challenge vial

Containing 100000 EU/vial shall be placed along with each sensor.

6.5.2.4 Detail of the challenge vial recorded as per Exhibit-E05.

6.5.2.4 Five Biological Indicators strips containing Bacillus atrophaeous ATCC No. 9372 having

population count not less than106 CFU shall also be placed in each cycle inside the article which

are kept in the coolest part of the chamber determined during Heat distribution study.

6.5.2.5 Details of each cycle run has been recorded as per Exhibit-E03.

6.5.2.6 The observation of Heat Penetration Study have been recorded as per Exhibit E04.

6.5.2.7 Details of the loads shall be as per Annexure-03.

6.5.2.8 Drawings showing location of the flexible RTDs in the chamber for heat penetration study


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shall be as per Annexure-04.

6.5.2.9 At the end of the cycle examined the measured temperature and noted the following:

The parts of the usable chamber space that are the fastest and the slowest to attain the

sterilization temperature.

The parts of the usable chamber space that are the Hottest and the coolest during the

sterilization holding time.

6.5.2.10 FD at a temperature other than 250°c is the time required to provide the lethality equivalent to

that provided at 250°c for a started time.

FD shall be calculated as follows

Where

∆T = Time interval between two successive measurements.

TA = Actual temperature at that point.

TB = Base temperature i.e. 121.1oC.

Z = 45(Change in temperature required to alter the D value by a factor of 10 i.e.1 log

Reduction).

6.5.2.11 Calculated the SAL values as per following formula and record the data as per Exhibit-E04.

FD

SAL = ------

D250

Where D250 is the time required to reduce the microbial population by 90% i.e. 1 log reduction.

6.5.2.12 D250 shall be calculated as follows:

TA-TB

------------

Z

D250 = D160 x 10

D250 is the D value at temperature 250°c.

D160 is the D value at temperature 160°c (Certified value).


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TA is 160°c

TB is 250°c

Z is the z value at 160°c.

6.5.3 CALCULATION OF FH VALUE

Definition of FH value (Lethal Rate / accumulated lethality):

The FH value is a "unit of lethality" and is defined as the number of minutes equivalent of

depyrogenation at reference temperature (250° C), calculated using a know Z value.

FH = dt x 10 (T-250) / Z

Where

dt: Interval of temperature monitoring in a certain location i.e. 01 minute.

T: Temperature at a certain time in certain location during depyrogenation hold

Z: Z Value which indicates relationship of lethality to temperature. (45ºC)

6.5.4 Acceptance criteria:

There should be temperature NLT 250°C at each probe in sterilizer chamber during

sterilization hold period.

Upper targeted limit for temperature will be 270°C. Based on validation study results for

upper limit will be finalized in report.

Minimum of 3 log reduction shall be achieved using Endotoxin challenge vials.

Minimum of 12 log reduction shall be achieved using Biological Indicators.

6.6 Endotoxin Challenge Study

Endotoxin Challenge Study is the part of Heat penetration study and shall be done for the set

load by placing endotoxin challenge vial next to each temperature sensor.

6.6.1 Take an Endotoxin challenge vial and prepare the Endotoxin dilution containing

100,000EU/ml and add 0.1 ml of 100000 EU/ml in each vial.

6.6.2 Take 18 No. of de-pyrogenated vials for each cycle to be challenged for depyrogenation.

6.6.3 Air dry the vials, seals them and mark them the respective location and cycle no.


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6.6.4 Place 16 vials containing 0.1ml of 10,000EU/ml along with each sensor inside the Chamber and

process at 250°C for not less than 30 minutes.

6.6.5 Keep two vials as such without processing them as positive control.

6.6.6 The Endotoxin challenge vial shall be placed along with each temperature sensor as per

Annexure-04

6.6.7 After processing let the vials cool to room temperature.

6.6.8 Reconstitute each processed vial and one unprocessed vial with 1ml of LRW and vortex for 15

min.

6.6.9 Take 0.1ml from each processed vial in reaction tubes in duplicate and add 0.1ml of lysate

(Sensitivity 0.125EU/ml).

6.6.10 For performing positive control prepare a dilution of 1:10,000 containing 1EU/ml of endotoxin

content from one unprocessed vial.

6.6.11 Take 0.1ml of 1EU/ml in reaction tubes in duplicate and add 0.1ml of lysate.

6.6.12 The details of the dilution preparation is as per Exhibit E06.

6.6.13 Perform Control curve from the unprocessed challenge vial also.

6.6.14 Incubate the Reaction tubes at 37±1°C for 60±2 minutes.

6.6.15 Details of the control curve have been recorded as per Exhibit E07.

6.6.15 Calculate the minimum log reduction using following formula

Minimum log reduction= log Endotoxin concentration of the unprocessed control vials- log

Endotoxin concentration of the processed vials

Where Endotoxin concentration = Lysate sensitivity x reconstituted volume x dilution

6.6.16 Observations:

- A positive reaction indicates Endotoxin content greater than the calculated Endotoxin value.

- A negative reaction indicates content less than the calculated Endotoxin value.

- A positive reaction in the 1/10,000 dilution of the unprocessed control vials indicates an initial


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Endotoxin concentration greater than 1250 EU/ml.

6.7 Non Viable Air Borne Particle Count

6.7.1 Objective

To demonstrate that the Dry Heat Sterilizer meets the requirement for desired cleanliness class

i.e. Class 100 (Grade A).

6.7.2 Procedure

a) This test shall be performed after cleaning of Dry Heat Sterilizer as SOP, “Operation and

Maintenance of Dry Heat Sterilizer.

b) Carry out the studies with Empty Chamber.

c) The temperature of Chamber of the Dry Heat Sterilizer should not be more than 5°C above

the ambient temperature during the study.

d) Ensure that blower of the Dry Heat Sterilizer is running and heater is OFF.

e) Perform the operation of Non-Viable Particle Counter at Centre (Triplicate Sampling) of the

dry heat sterilizer.

Acceptance Criteria:

The number of particles shall not increase more than the limits mentioned below.

• The observations of nonviable particle count shall be recorded as per Exhibit-E09 and test

reports shall be attached in Annexure-05.

6.8 Any deviation observed during performance qualification shall be recorded in the observed

deviation, corrective action and justification report section.

6.9 Observed deviation shall be reported to the department head and quality head.

6.10 If the observed deviation does not have any major impact on the qualification the final conclusion

shall be provided.

Class Max. No. of permitted particles Per cubic Meter (At Rest)

≥ 0.5 µm ≥ 5.0 µm

100 3520 29


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6.11 If the observed deviation has major impact on the qualification, deviation shall be reported to the

manufacturer for the corrective action and qualification activity shall be performed again.

7.0 Acceptance Criteria

Performance qualification shall be considered acceptable when requirements listed in section 6.0 of

this protocol has been fulfilled and all the components of Dry Heat Sterilizer, Fully Automatic

are performing as per our requirement and as per manufacturer recommendations.

8.0 Qualification Report

The performance qualification report shall consist of a summary document, in narrative form,

which shall briefly describe the activity performed along with the observations recorded in

relevant exhibits.

This report shall also include the related documents and attachments/annexure which were

completed at the time of qualification activity.

9.0 Approval of Qualification Report

The report shall be evaluated and proper references / conclusions / recommendations shall be

recorded by quality assurance.

The performance qualification report shall be evaluated and finally approved by quality

assurance.


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10.0 Observed Deviation

S.No. Page No. Point

No.

Observed Deviation Deviation

Reported

By

Deviation

Approved By

Corrective Action Taken Justification of Corrective

Action

Corrective action taken

and justification given by

Report Approved By

Department Head Quality Head


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11.0 List of Exhibits/Annexure

11.1 List of Exhibits

Exhibit No. Exhibit Title No. of Pages

E01 Calibration Certificate Availability Verification Checklist

E02 Calibration and Temperature Verification Checklist

E03 Heat Distribution Study

E04 Heat Penetration Study

E05 Details of Endotoxin Challenge Vial

E06 Dilution Preparation for Endotoxin Challenge Study

E07 Detail of Control curve of Endotoxin Challenge Vial

E08 HEPA Filter Integrity Testing

E09 Non-viable Air Borne Particle Count

E10 Air velocity Test

E11 Air Flow Pattern Test

Total No. of Pages

11.2 List of Annexure

Annexure No. Annexure Title No. of Pages

01 Training Record

02 Drawing of Locations of Temperature Probes in the

Chamber for Heat Distribution Study

03 Details of the Sterilization Loads

04 Drawing of Locations of Temperature Probes in the

Chamber for Heat Penetration Study

05 Test Reports

06 Calibration Certificates

07 Print outs of the Data

Total No. of Pages

12.0 Reference Documents

➢ ISO 14644 of Clean Rooms and Associated Controlled Environments.

➢ Validation Master Plan.

➢ Schedule - M – “Good Manufacturing Practices and Requirements of Premises, Plant and

Equipment for Pharmaceutical Products.”


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Exhibit – E01

Calibration Certificate Availability Verification Checklist

Equipment Name / Description : Dry Heat Sterilizer, Fully Automatic

Tag No . : QB/DHS/001

Location : Microbiology Lab (Ground floor)

S.No. Document

Description

Specifications Actual Observations

1. Calibration Certificates of all the

instruments attached

All the calibration

certificates should

available

Remarks: Observations Comply / Not Comply with the specifications.

Checked By:____________________ _________ ________

(Name) (Sign) (Date)

Verified By:____________________ _________ ________


Exhibit – E02


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Calibration and Temperature Verification Checklist

Location :

Set Temperature:

Pre calibration/Post Calibration study started at:

Probe Description Temperature Observed (ºC)

Probe 1

Probe 2

Probe 3

Probe 4

Probe 5

Probe 6

Probe 7

Probe 8

Probe 9

Probe 10

Probe 11

Probe 12

Probe 13

Probe 14

Probe 15

Probe 16

Acceptance Criteria – Temperature difference among probe to probe shall not be more than

±0.5ºC.

Remarks: _______________________________________________________________

Checked By: ____________________ _________ ________


Verified By: ____________________ _________ ________



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Exhibit – E03

Heat Distribution Study

Date of test

Cycle No.

Type of load

Set sterilization temperature

Set sterilization period

Process cycle started at Hrs.

Dwell period started at Hrs.

Dwell period ended at Hrs.

Temperature profile

Temp. Sterilization temperature of different sensors

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Max.

Min.

Avg.

Endotoxin

reaction of

vials

Observed

log

reduction

+: Denote gel clot formation -: Denote no gel clot formation


-There should be uniform distribution of temperature NLT 250°C at each probe in sterilizer chamber

during sterilization hold period. Upper targeted limit for temperature will be 270°C.

-Minimum 3 log reduction shall be achieved at each location while doing Endotoxin challenge study.

Remarks: Dry heat sterilizer complies/does not comply with the above acceptance criteria.

Checked By: ____________________ _________ ________


Verified By: ____________________ _________ ________



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Exhibit – E04

Heat Penetration Study

Date of test

Cycle No.

Type of load

Set sterilization temperature

Set sterilization period

Process cycle started at Hrs.

Dwell period started at Hrs.

Dwell period ended at Hrs.

Temperature profile

Temp. Sterilization temperature of different sensors

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

FD Value

SAL

Endotoxin

reaction

of vials

Endotoxin

log

reduction



-There should be temperature NLT 250°C at each probe in sterilizer chamber during sterilization hold

period. Upper targeted limit for temperature will be 270°C.

- Minimum of 3 log reduction shall be achieved using Endotoxin challenge vials.

- Minimum of 12 log reduction shall be achieved using Biological Indicators.

Remarks: Dry heat sterilizer complies/ does not comply with the above acceptance criteria.

Checked By: ____________________ _________ ________


Verified By: ____________________ _________ ________



PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Exhibit – E05

Details of Endotoxin Challenge Vials

Make

Catalogue No.

Lot No.

Potency

Volume to be reconstituted

Expiry date

Checked By:____________________ _________ ________


Verified By:____________________ _________ ________



PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Exhibit-E06

Dilution Preparation for Endotoxin Challenge Study

LAL Reagent Details

Manufacturer : Lot No. : Expiry Date :

Sensitivity :

LAL Water Details

Lot No. : Expiry Date : Date of

Opening :

Dilution Preparation:

Checked By:____________________ _________ ________


Verified By:____________________ _________ ________



PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Exhibit-E07

Detail of Control curve of Endotoxin Challenge Vial

Batch No/Lot No of LAL : ______________________ Date of Test : ________________

Incubation Temperature: 37 ± 1ºC Incubation Period: 60 ± 2 min.

LAL reagent details

Manufacturer: Expiry date:

Sensitivity: Reconstituted on: Use before:

LAL regent water details

Lot No: Expiry date: Vial opened on:

Test Description Tube 1 Tube 2 Inference

Blank

Control 2λ (_______EU/ml)

Control λ (_______EU/ml)

Control λ/2 (_______EU/ml)

Control λ/4 (_______EU/ml)

Geometric mean( GM):

∑ e ∑ e= Summation of all endpoints

GM= antilog ------------- f= number of replicates

f

=

=

Remarks: Sensitivity of LAL reagent is within/not within ± two fold dilution.

Acceptance criteria: LAL regent qualifies if GM confirms the label claim sensitivity of the reagent

within ± two fold dilution.


Checked By:____________________ _________ ________


Verified By:____________________ _________ ________



PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Exhibit – E08

HEPA Filter Integrity Testing

Equipment ID No._____________________ Date of test____________________________

Name of the Instrument used for test____________________________________________________

Make_______________________________ Model________________________________

Date of calibration____________________ Due date of calibration___________________

Sr. No._____________________________ Concentration of PAO upstream____________

Filter No. Maximum Downstream

Concentration of PAO observed Remarks

Acceptance criteria: NMT 0.01% Poly alpha Olefin shall penetrate across the HEPA filter and no

leakage shall be observed from the fitment.

Remarks: ________________________________________________________________________

_________________________________________________________________________________

Checked By: ____________________ _________ ________


Verified By: ____________________ _________ ________


Exhibit – E09


PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Non Viable Air Borne Particle Count

Date of test__________________________________ Filter No.________________________________

No. of locations to be sampled__________________ Witnessed By_____________________________

Instrument used for test________________________ Make___________________________________

___ Model______________________________________ Sr. No___________________________________

Date of calibration____________________________ Due date of calibration______________________

Volume of air sampled ________________________ Time for which air sampled__________________

Location Particle count 0.5μ per m3 Particle count 5.0μ per m3

1 2 3 Avg. 1 2 3 Avg.

L1

L2

L3

L4

L5


Class100: Particles ≥ 0.5 μm =NMT 3520 , Particles ≥ 5.0 μm = NMT 29

Conclusion: Air sampled complies/does not comply with as per Acceptance criteria

Checked By: ____________________ _________ ________


Verified By: ____________________ _________ ________


Exhibit-E10

Air velocity


PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Date of test__________________________

Name of the Instrument used for test______________________________________________________

Make_______________________________ Model___________________________________

Date of calibration____________________ Due date of calibration______________________

Sr. No._____________________________

Location Air velocity (FPM)

V1

V2

V3

V4

V5

Average in FPM

Witnessed by

Acceptance Criteria: The Average air velocity shall be within 90 ± 20% FPM.

Remarks: Results are within/ not within limit.

Reviewed By:____________________ _________ ________

(QC) (Name) (Sign) (Date)

Verified By:____________________ _________ ________

(QA) (Name) (Sign) (Date)

Exhibit- E11

AIR FLOW PATTERN TEST


PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Instrument

Name

Date of Test

Make Instrument ID.

Date HEPA filter Location Acceptance Criteria Observation

Observed

By

QA

Sign/Date

Right Hand Side HEPA

Filter

Fumes should follow the path from

supply air filter face to the grill of

return air riser under the chamber.

Left Hand Side HEPA

Filter

Acceptance criteria: The smoke flow should be unidirectional from the HEPA filter face to return air

riser.

Remarks: Air flow direction Comply/does not comply as per the acceptance criteria.

Checked By: ________________________ ___________ ____________


Verified By: ________________________ ___________ ____________



PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Annexure – 01

Training Record

Equipment Name: Dry Heat Sterilizer ( Fully Automatic)

Equipment Tag No.:

Location: New Microbiology Lab

No. of Pages:


PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Annexure – 02

Drawing Of Locations of Temperature Probes in the Chamber

for Heat Distribution Study


Equipment Tag No.:


No. of Pages:


PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Annexure – 03

Details of the Sterilization Loads


Equipment Tag No.:


No. of Pages:


PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Annexure – 04

Drawing of Locations of Temperature Probes in the Chamber for Heat

Penetration Study


Equipment Tag No.:


No. of Pages:


PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Annexure – 05

Test Reports


Equipment Tag No.:


No. of Pages:


PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Annexure – 06

Calibration Certificates


Equipment Tag No.:


No. of Pages:


PROTOCOL

FOR

DRY HEAT STERILIZER

SUPERSEDES NIL

EFFECTIVE DATE



Annexure – 07

Print Outs of the Data


Equipment Tag No.:


No. of Pages: