REVITAL HEALTHCARE (EPZ) LTD Document No.

: Z/F/06/14D
Revision No.: 00
STERILIZATION PROCESS VALIDATION Effective Date: 19/04/2022

ETO STERILIZATION
PERFORMANCE QUALIFICATION REPORT
PRODUCT: 0.5ML SYRINGE
EQUIPMENT NO.: STL/MC/06

Report No: PQR01010622

EFFECTIVE DATE: - 19/04/2022

NEXT DUE DATE: - 19/04/2025

ETO Sterilizer Make: HUAQING

Equipment ID: STL/MC/06
Capacity: 15 M3/15034Ltr.
Operator Name: Mr. Jonathan Safari
Supervisor Name: Mr. Robert Mutua
Department (Place): Sterilization Area

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REVITAL HEALTHCARE (EPZ) LTD Document No.: Z/F/06/14D
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REVISION HISTORY
Revision no. Revised Page(s) Description Date
00 00 Original Issue 19/04/2022

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TABLE OF CONTENTS

S. No. Description Page No.

1.0 Report approval 4

2.0 Responsibilities 5

3.0 Purpose 5

4.0 Scope 5

5.0 Equipment Description 6

6.0 Sterilizing agent characterization 6

7.0 Product and packaging description 6

8.0 Calibration 7

9.0 Process qualification 7

10.0 Validation summary 13

11.0 Revalidation 14

12.0 Personnel 14

13.0 14

14.0 Reference document 15

15.0 Conclusion/recommendation 15

1. REPORT APPROVAL SHEET

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NAME DESIGNATION SIGNATURE DATE

Prepared by
QA Assistant 19/04/2022

Reviewed by
Manager QC 19/04/2022

Reviewed by
Manager QA 19/04/2022

Reviewed by
Production manager 19/04/2022

Approved by
HOD (QC/QA) 19/04/2022

Approved by
Operation Director 19/04/2022

2.0 RESPONSIBILITY
Following persons are responsible for the validation & routine control:

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Sterilizer operators
i. To physically load and offload the sterilizers.
ii. Placement of BIs and post treatment of the same.
Production designee
i. To make available the required quantity of the product.
Quality assurance.
i. Preparation of protocol and report
ii. Review and approval of protocol and report.
iii. Execution of validation activity as per the approved protocol.
Quality Control
i. Initial bio-burden of the product.
ii. Post treatment sterility testing
Trucal services
i. Adequate qualification of equipment and calibration of instrument and machine prior to
execution of the validation study.
Accuprec Labs
ETO residue testing of worst case samples post treatment.
3.0 PURPOSE: -
The report has been evaluated and confirmed the criteria for the process validation of
15m³ sterilizer chamber volume, utilizing ethylene oxide gas, for the sterilization of
medical devices manufactured in Revital Healthcare (EPZ) Limited. It has established
and documented the sterilization validation and demonstrated that the cycle as described,
will repeatedly deliver the sterility assurance level (SAL) of 10-6 to the product when
processed using the validated operating conditions presented in the validation summary
report. This report is designed to be consistent with ISO 11135-1 – Sterilization of health
care products – Ethylene oxide – Part1: “Requirements for development, validation and
routine control of a sterilization process for medical devices”.

4.0 SCOPE: -
This report is limited to validation of sterilizer located at Revital Healthcare (EPZ)
Limited (RHEL), the ETO sterilizer machine No. STL/MC/06, a 15 m³ chamber
measuring L 670cm X W132cm X H170cm.
The scope of the validation will be applicable to 0.5ml AD Syringe as a representative of
other products manufactured and sterilized at Revital Healthcare (EPZ) LTD using
sterilizer No.6. The STL/MC/06 has a capacity of 80 cartons for 0.5ml 2Part AD Syringe
and whose product density is 8.074g/cm3.

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Physical and Microbial PQ was performed with 0.5ml 2P AD Syringe.

This report is limited to validation and qualification of STL/MC/06 located at RHEL for 0.5ml
AD Syringe.

5.0 EQUIPMENT DESCRIPTION

i. ETO sterilizer – 15m3 in volume
ii. Vaporizer
iii. Vacuum pump
iv. ETO filter
v. Gas cylinder containing ETO gas
vi. Weighing scale
vii. Hot water generating and circulating unit
viii. Steam generating unit
ix. Extra temperature and humidity indicators
x. Biological indicators

6.0 STERILIZING AGENT CHARACTERIZATION.

The Ethylene Oxide gas used for sterilization is of the following concentration: ETO
100% (20:80 for Ethylene-oxide: Carbon dioxide).

7.0 PRODUCT AND PACKAGING DESCRIPTION.

7.1. Product for validation
This validation exercise is being carried out for 0.5ml 2P AD Syringe.

7.2. Packaging of the product

The products are packed in paper poly pouch which consists of medical grade paper for
primary packaging, an inner box contains 100 pcs of syringes for secondary packaging
and 3600 pcs in shipper cartons for tertiary packaging.

7.3. The validation loads.

80 cartons of the product formed one sterilizer cycle load. Five such loads were provided,
thus 400 cartons were utilized for the whole validation exercise. Following is the details
of the products used for the validation exercise:

0.5ml 2piece AD syringe

Product raw material Product batch/lot No. of cartons Quantity in

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details number pieces

Barrel- polypropylene 030922 160 576000
Plunger- high density
polyethylene 031022 160 576000
031122 80 288000

8.0 CALIBRATION.
All controlling, measuring and indicating instruments were calibrated according to their
calibrating system identified in the company procedure manuals. The calibration
results/records were reviewed before the commencement of the validation.

9.0 PROCESS QUALIFICATION

The process qualification was conducted once the sterilizer was calibrated and the
ancillary instruments successfully installed. The report for sterilizer qualification consists
of the following sub steps:

9.1. INSTALLATION QUALIFICATION (IQ)

IQ demonstrated that the sterilization equipment and any ancillary items have been
supplied and installed in accordance with their specification. Drawing and description of
the critical ancillary systems have been documented in the IQ Report (IQR). The
calibration records were checked and verified.

9.2. OPERATIONAL QUALIFICATION (OQ)

Prior to OQ, the calibration of all instrumentation used for monitoring, controlling,
indicating or recording was confirmed. OQ demonstrated that the installed equipment is
capable of delivering the specified process within defined tolerances.
Sterilizer qualification report has been documented in the OQ Report (OQR), number
OQR010620. The conclusions for the same along with the Protocol and Report are
attached as Annexure 9

9.3. PERFORMANCE QUALIFICATION – PHYSICAL (PPQ)

The product for PPQ was packed in an identical manner to that when routinely presented
for sterilization. In this case 80 cartons were loaded as per the loading pattern Annexure-
01. The following additional instruments were used for the PQ:
i. 15temperature sensors and 6 humidity sensors.
ii. ETO gas cylinders.

One simulation trial was carried out wherein the following steps in the sterilization
process were monitored:

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9.3.1. Proforma for chamber leak-rate:

The vacuum till the set value of -0.63±0.01 kg/cm³ was induced. The vacuum
level was maintained for 10min. Dip in vacuum for 9min was monitored. The leak
rate was determined for 90 minutes.
There is no expected occurrence of chamber leak. In case of chamber leak, the
allowed tolerance for a pressure change during leak test is -0.63kg/cm³ /10
minutes, which implies that the sterilizers are performing as per the standard.
The data for the chamber leak trial is attached as Annexure 12

9.3.2. Temperature Profile of loaded sterilizer:

15temperature sensors and 6 humidity sensors were inserted into the sterilizer and
fixed at various positions in the product load. These positions were noted and
enclosed in Annexure 7.
The sterilizer door was closed tightly. Hot water circulation for heating the
sterilizer. Scanner readings were noted at every 10 minute. The min and max
temperatures were noted down. The temperature difference is also calculated to
check if it was in the accepted range of ±10°C.

Temperature readings (°C)

Max. Min. Difference.
54.9 45.1 9.8

9.3.3. Pre-Exposure evacuation:

The initial pressure in the sterilizer was recorded. The vacuum pump was
switched on and the vacuum was drawn up to -0.63kg/cm³, which is the set value.

9.3.4. Steam Injection:

6 humidity sensors introduced were geometrically placed within the load to
capture maximum variation in the sterilizer, Annexure 7. The steam generator
was started and steam injected after the vacuum was drawn up to -0.62 kg/cm³.
The steam valves were opened and the time noted.
As soon as the vacuum was reduced to -0.61 kg/cm³, the steam valve was closed.
Vacuum was again drawn up to -0.60 kg/cm³ and then steam injected till -0.64
kg/cm³. The humidity readings during a humidification dwell time of 10 minutes
were recorded. The data for this was tabulated from the output given out by the 6
Humidity sensors.
The humidification time is obtained from the PLC output (controlling output)

During Exposure
Max (%) Min (%)
59.8 30.1

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9.3.6 Sterilant injection

The weight of the sterilant cylinder on the weighing scale was recorded. The
sterilant injection in the sterilizer was started and initial time was noted. The
injection was continued till 28.5 of the gas was released in the sterilizer as
indicated on the weighing scale on the LED display. EO Valve auto shut down
after the injection of the 28.5kg of EO GAS was checked. The time taken for
injection of sterilant was recorded and also the dip in vacuum was noted.

Batch Weight of Initial Pressure Time Temperature of

No. EtO pressure after taken sensor measuring
introduced (kg/cm3) EtO (min) EtO Entry point
(kgs) Injection (oC)
(kg/cm3)
Min. Max.
03092 28.5 0.02 1.06 17 45.1 54.8
2
03102 28.5 0.02 1.06 15 45.1 54.8
2
03112 28.5 0.02 1.06 16 45.1 54.8
2

9.3.7 Post Exposure Evacuation & Aeration:

The vacuum was drawn till the set value of -0.67 kg/cm³. Once the vacuum level
was attained, the same was maintained by ETO injection at 1.02 kg/cm³. The
above steps of evacuation and ETO injection were repeated followed by an
evacuation up to -0.03 kg/cm³.
Aeration: The air valves were then opened immediately after the last evacuation
such that filtered air was continuously allowed into the sterilizer as well as drawn
out by vacuum creating an air cycle. This was continued for 60 minutes.

9.4 PERFORMANCE QUALIFICATION-MICROBIOLOGICAL

This validation was conducted according to the protocol number PQP01010622 which is
designed to incorporate elements of half-cycle validation methods as outlined in the standard
ISO 11135 Sterilization of healthcare products-ethylene oxide-requirements for the development,
validation and outline control of sterilization process for medical devices. The method involved
is the Overkill approach.
Half-Cycle Qualification:
This method determined the minimum time of exposure to ethylene oxide with all other process
parameters except time remaining constant, at which there were no survivors. Two further

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experiments were also performed to confirm the minimum time. All trials showed no growth in
the biological indicators. The specified exposure time will be at least double this minimum time.
In each cycle, 35 biological indicators containing about 106 viable spores of Bacillus atrophaeus
ATCC9372, were used which comply with AAMI/ISO 11138.
9.4.1. Fractional cycle
The microbiological trials were conducted by using 80 boxes for the fractional cycle batch. The
35 BI’s (IPCD) were placed inside 35 shipper cartons. These cartons were then loaded into the
sterilizer. Additionally, 35 Chemical Indicators were also placed (EPCD) on the outside of the
cartons. The placement is enclosed in annexure 8 – BI location. On all the cartons, chemical
indicators (stickers) were also pasted.
For fractional cycle trial, the batch report is enclosed as Annexure 13.
The summary of the fractional cycle is as follows:
Batch No. 030922
PARAMETERS OBSERVATIONS
Loading Time 15±5 min.
No. of chemical indicator 80
No. of Biological indicators 35
Minimum temperature of product
27-30˚C
entering the sterilization process
Pre- conditioning Temperature 45-55˚C
Pre- conditioning Time 60Minutes
Pre- conditioning Humidity 30-60 %
Initial Vacuum 0.02 Kg/cm2
Initial Vacuum hold time 10 Minutes
Conditioning time 15 minutes
Gas Injection/ Charging pressure -0.67-1.06kg/cm2
EO Injection time ≤20 min
Volume of EO used 28.5 kg
Gas Exposure/ Hold time 90 minutes
Cycle temperature during exposure/
45-55˚C
hold
Relative Humidity During Exposure 30-60%
Gas Pressure during Exposure/hold 1.06 kg/cm2
No. of air wash 05
Air wash vacuum -0.60±5 kg/cm2
Data recording time during Exposure 10 minutes
Aeration-1 10±1min
Aeration-2 8±1min
Aeration-3 60±5min
Temperature during Aeration/ Air 45-55˚C

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wash
Unloading Time 15±5min

9.4.2. Half Cycles

Three successful cycles were performed using 35 biological indicators as IPCD, 35 Chemical
Indicators as EPCD as per protocol for the same load configuration. On all the cartons, chemical
indicator (stickers) were also pasted.
For half cycle’s trial, the batch report is enclosed as Annexure 14.
The summary of the half cycles is as follows:
Batch No. 030922,031022
PARAMETERS OBSERVATIONS OBSERVATIONS OBSERVATIONS
Loading Time 15±5 min. 15±5 min. 15±5 min.
No. of chemical indicator 80+35 80+35 80+35
No. of Biological
35 35 35
indicators
Minimum temperature of
product entering the 27-30˚C 27-30˚C 27-30˚C
sterilization process
Pre- conditioning
45-55˚C 45-55˚C 45-55˚C
Temperature
Pre- conditioning Time 60Minutes 60Minutes 60Minutes
Pre- conditioning
30-60 % 30-60 % 30-60 %
Humidity
Initial Vacuum 0.02 Kg/cm2 0.02 Kg/cm2 0.02 Kg/cm2
Initial Vacuum hold time 10 Minutes 10 Minutes 10 Minutes
Conditioning time 15 minutes 15 minutes 15 minutes
Gas Injection/ Charging
-0.67-1.06kg/cm2 -0.67-1.06kg/cm2 -0.67-1.06kg/cm2
pressure
EO Injection time ≤20 min ≤20 min ≤20 min
Volume of EO used 28.5 kg 28.5 kg 28.5 kg
Gas Exposure/ Hold time 180 minutes 180 minutes 180 minutes
Cycle temperature during
45-55˚C 45-55˚C 45-55˚C
exposure/ hold
Relative Humidity During
30-60% 30-60% 30-60%
Exposure
Gas Pressure during
1.06 kg/cm2 1.06 kg/cm2 1.06 kg/cm2
Exposure/hold
No. of air wash 05 05 05
Air wash vacuum -0.60±5 kg/cm2 -0.60±5 kg/cm2 -0.60±5 kg/cm2
Data recording time
10 minutes 10 minutes 10 minutes
during Exposure

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Aeration-1 10±1min 10±1min 10±1min

Aeration-2 8±1min 8±1min 8±1min
Aeration-3 60±5min 60±5min 60±5min
Temperature during
45-55˚C 45-55˚C 45-55˚C
Aeration/ Air wash
Unloading Time 15±5min 15±5min 15±5min

9.4.3. Full Cycle

One successful full cycle was performed with an exposure time of six hours which is double the
half cycle exposure time for the same load with 35 biological indicators as IPCD and 35
Chemical Indicators as EPCD. On all the cartons, chemical indicator (stickers) was also pasted.
For full cycle trial, the batch report is enclosed as Annexure 15.
The summary of the full cycle is as follows
Batch No. 031122
PARAMETERS OBSERVATIONS
Loading Time 15±5 min.
No. of chemical indicator 80
No. of Biological indicators 35
Minimum temperature of product
27-30˚C
entering the sterilization process
Pre- conditioning Temperature 45-55˚C
Pre- conditioning Time 60Minutes
Pre- conditioning Humidity 30-60 %
Initial Vacuum 0.02 Kg/cm2
Initial Vacuum hold time 10 Minutes
Conditioning time 15 minutes
Gas Injection/ Charging pressure -0.67-1.06kg/cm2
EO Injection time ≤20 min
Volume of EO used 28.5 kg
Gas Exposure/ Hold time 360 minutes
Cycle temperature during exposure/
45-55˚C
hold
Relative Humidity During Exposure 30-60%
Gas Pressure during Exposure/hold 1.06 kg/cm2
No. of air wash 03
Air wash vacuum -0.60±5 kg/cm2
Data recording time during Exposure 10 minutes
Aeration-1 10±1min
Aeration-2 8±1min
Aeration-3 60±5min
Temperature during Aeration/ Air 45-55˚C

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wash
Unloading Time 15±5min
9.4.4. The biological indicators were retrieved after all the sterilization cycles and incubated for
seven days in the incubator at Revital microbiology lab. The biological indicator viability reports
enclosed as Annexure 11
All the Chemical Indicators were handed over to Chemical lab to check the Color change. All the
sticker labels which were pasted outside of the cartons were also checked for change in Color.
(Refer Annexure 11)
Product Cycles subjected to Purpose
Group 1 One Fractional cycle One Additional Full Worst case
Cycle
Group 2 Three half cycles Three additional half Full Cycle
cycle
Group 3 One Full cycle ---------- Full Cycle

9.4.5. The bio-burden reports of the untreated products have been attached as Annexure 10
9.4.6. The sterility reports of the products treated in the one Fractional cycle, three Half Cycles,
and one Full cycles have been enclosed as Annexure 16
10. VALIDATION SUMMARY
10.1 The physical qualification study performed has met the required specifications as
follows:
10.1.1 Throughout the exposure time, the sterilization load attained the minimum
specified temperature range of 50±5oC and the temperature range across the
product load was ±5oc at any given time during the sterilant exposure.
10.1.2 The RH distribution throughout the load was greater than 30% i.e. 45±15%
10.1.3 Time taken to achieve the vacuum of -0.60 kg/cm³ was 10 minutes.
10.1.4 There was no leakage observed in the chamber.
10.1.5 Time taken for ETO gas injection was ≤20minutes
10.1.6 Time taken for the admission of 28.5kg of sterilant gas in the sterilizer was
≤20minutes
10.1.7 Time taken for post exposure evacuation & aeration is roughly 60 minutes
10.2 The microbiological qualification obtained a SAL 10 -6 as the following criteria were
met:

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10.2.1 The biological indicators used during half and full cycles showed no
growth upon 7 days incubation.
10.2.2 The Chemical Indicators used during fractional, half and full cycles shown
Colour change from brown red to yellow.
10.2.3 All other chemical indicators pasted on the outside of the carton shown
Colour change from Reddish brown to green.
10.2.4 The residual level gases are within the limits as specified i.e ethylene oxide
(EO) 1ppm, ethylene chlorohydrin (ECH) 50ppm and ethylene glycol (EG)
620ppm. Annexure 18
10.2.5 The sterility test carried out on products treated in the half cycles, full
cycles and fractional cycle passed. Annexure 16
10.2.4 The product used during validation met all performance criteria as
stipulated by Revital healthcare (EPZ) LTD.
10.3 The process parameters to be used for normal commercial batches are attached as
Annexure 3

11. REVALIDATION
Revalidation shall be performed under the following conditions:
11.1 New Sterilizer
11.2 New facility
11.3 Change in validated process parameters
11.4 Any change identified to have direct impact on the sterilization process
11.5 Customer/regulatory authority requirement
11.6 Routine revalidation once in three years
11.7 New product introduced for which equivalence with validated product cannot be
established.
12. PERSONNEL
The personnel required for conducting the validation trials are sufficiently trained in the
validation process and operation and their individual records are available.
13. PRODUCT RELEASE AFTER STERILIZATION

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The product three lots was used during the fractional cycle, half cycles and full cycle
(400cartons).
The number of cartons used during the fractional cycle was also utilized an additional full
cycle. The number of cartons treated in the 3 half cycles was utilized to conduct the 3
other half cycles, one each to complete the cycles to full cycles.
The performance characteristics of these products were tested Revital healthcare (EPZ)
Limited.
14. REFERENCE DOCUMENTS:
14.1. ISO 11135, Sterilization of health-care products –Ethylene oxide - Requirements
for the development, validation and routine control of a sterilization process for medical
devices.
14.2. ISO 11138-2, “Sterilization of health care products - Biological indicators - Part 2:
Biological indicators for ethylene oxide sterilization processes”
14.3. ISO 10993-7: Biological evaluation of medical devices – Part 7: Ethylene oxide
sterilization residuals.
15. CONCLUSION/RECOMMENDATION:
The results of the validation were found to be complying with the acceptance criteria
detailed in protocol. Thus STL/MC/06 stands validated for sterilizing Revital’s various
varieties of hypodermic syringes. O.5ml 2piece AD Syringe among other syringes
manufactured at Revital are made and packed using the same Raw and Packaging
material, therefore, 0.5ml 2piece AD Syringe which has the highest density is a
representative of all other syringes.

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