PQ Protocol - Final

Medivac Surgical Pvt. Ltd.-Kutch, Gujarat India. Doc No : MVS/QP/19.
0/A-06
Revision No : 0.0
PROCESS VALIDATION - ETO STERILIZER
Effective Date : 01-06-2018
F88 PERFORMANCE QUALIFICATION OF STERILIZER
EO STERILIZER- MSPL/ETO/01
<<<X<
1. APPROVAL OF PERFORMANCE QUALIFICATION PROTOCOL
The protocol has been prepared, checked, and approved by the concerned personnel for
implementation by the undersigned:
DEPARTMENT NAME DESIGNATI SIGNATURE DATE

PREPARE ON
D BY
Quality
Gaurav Microbiologist
Assurance
joshi
DEPARTME NAME DESIGNATI SIGNATURE DATE

NT ON
Alpesh
AGM
Production Ghoghari
CHECKED
BY Utility Jayesh Pandya In-Charge
Plant & Aadish

Director
Project Aggarwal
DEPARTMENT NAME DESIGNATION SIGNATURE DATE

APPROVE
D BY
Quality Manager
Viral Sheth
Assurance
Page No Prepared By Reviewed By Approved By

Gaurav Joshi Alpesh Ghoghari Viral Sheth
1 of 34
Medivac Surgical Pvt. Ltd.-Kutch, Gujarat India. Doc No : MVS/QP/19.0/A-06
Revision No : 0.0
<
2. OBJECTIVE:
The objective of performance qualification of EO sterilizer is as under:
 The objective of validation is to documents the evidence required to provide a high degree of assurance
that a specific process will consistently produce product meeting the required sterility assurance level (SAL).
Product sterilized in the validated process should be shown to meet predetermined specification and quality
characteristics related to product functionality and safety.
 To determine that Ethylene oxide sterilization process consistently performs as intended by running the
system and recording all relevant information. The Data and Test results must demonstrate that the
process meets pre-determined specifications under normal conditions as well as worst case conditions.
 Performance qualification of rigorous microbiological and physical testing, beyond routine monitoring, to
demonstrate the efficacy and reproducibility of the sterilization process.
 The results of the equipment validation shall meet pre-determined acceptance criteria(s) under normal
conditions of operations, and where appropriate for worst-case situations. If any variation is observed
between the set acceptance criteria and the actual data obtained, then same should be explained
reasonably.
 On the basis of validation data, the Work Instructions (WI) for routine monitoring of sterilization shall be
evaluated for its suitability.
In case any deviation is observed during validation, it shall be recorded in Deviation Section.
 The site Quality Assurance In-charge shall decide whether deviation is acceptable or not. If deviation is
acceptable and it does not impact the quality and Performance of sterilizer, continue the validation. If
deviation is not acceptable and it has impact on the quality and Operational performance of sterilizer,
necessary corrective action shall be taken before qualification.
 The equipment conforms to the complete qualification requirements and shall permit a formal “RELEASE” of
the equipment for the routine use in sterilization process.
<<<<X<
3. SCOPE:
The scope of this validation is as under for Sterilization of
 3 Way stop cock with/without extension tube  Infusion (IV) Set / Transfusion (BT) set
 I.V. Flow regulator  Measured Volume Infusion Set
 I.V. Cannula  Extension Line/ Pressure Monitoring Line
 Hypodermic Syringes  Hypodermic Needle
 Other similar packaged products 
The detail of equipment is as under:

2 of 34
Revision No : 0.0
Name of the Equipment : EO (Ethylene Oxide) Sterilizer- 01
Technical Identification No. : MSPL/ETO/01

Hangzhou Unique disinfection equipment co., Ltd.
Equipment Make :
Model No. : HDX-EN-20
Inner container volume : 20 m3
Equipment’s Location : Ground floor near Finished Goods.
4. RESPONSIBILITY
DEPARTMENT DESIGNATION RESPONSIBILITY
Shall prepare validation Protocol & report as

Quality Assurance Microbiologist per the guideline of relevant SOP and
validation procedure.
An execution team shall be made by A.G.M.
QA to execute the validation as per the
protocol and analyse the data and
preparation of validation
report.
Maintenance, Shall review the validation protocol & report
Assistant Manager /
Production, & for correctness, completeness, and technical
Deputy Manager/
Quality excellence.
Manager/ A.G.M.
Control, Quality
Assurance
Shall approve the validation protocol &

Quality Assurance Manager
report for correctness and completeness

3 of 34
Revision No : 0.0
<X<
5. MATERIALS, INSTRUMENTS AND DOCUMENTS
S. Name of Material
No.
Three way Stop Cock
1. Packed in blister (unit container) made of medical grade paper and PP+PE film. Further
unit pack is packed in duplex of 50 Blister Packed three-way stop cock and finally these
duplex boxes are packed in Corrugated box.
2. Calibrated Temperature & RH sensors
3. Validation procedure (QP/QAD/25, Rev. No.-09)

Routine Operation And Monitoring of ETO Sterilization Cycle For I.V. Cannula, 3 Way Stop
4.
Cock & 3 Way Extension Tubes (WI/IVC/25)
5. Biological indicator (Bacillus Atropheous, Population-106)
6. Chemical indicator Tape
7. Ethylene oxide gas cylinder (90% EO & 10% CO2)
8. Calibrated Weighing balance
9. Bio burden on product (GTP/QC/12)
10. Population verification of Biological indicators
11. EO Sterilizer
6. REFERENCE INTERNATIONAL STANDARDS USED FOR VALIDATION
 ISO 11135:2014/ EN ISO 11135-1: 2015, Sterilization of health care products- Ethylene oxide- - Requirements
for development, validation and routine control of a sterilization process for medical devices
 ISO 10993-7:2008, Biological evaluation of medical devices- Part 7: Ethylene oxide sterilization residuals
 BS EN ISO 11737-1:2006, Sterilization of medical devices - Microbiological methods -Part 1:

Determination of a population of microorganisms on products
 BS EN ISO 11737-2: 2009, Sterilization of medical devices - Microbiological methods - Part 2: Tests of
sterility performed in the validation of a sterilization process
 ISO 14161 : 2009, Sterilization of health care products —Biological indicators — Guidance for the
selection, use and interpretation of results

4 of 34
Revision No : 0.0
<<<<X<
7. SYSTEM/ EQUIPMENT DESCRIPTION - Brief description of equipment (sterilizer)
The Ethylene oxide sterilization process uses ethylene oxide as a sterilant to destroy viable microorganisms.
The ethylene oxide sterilization is suitable for the products which are heat and moisture sensitive.
The sterilizer (ETO-01) is having inner dimension of chamber as under:
Length: 9.0 meter

Width: 1.35 meter
Height: 1.7 meter
Sterilizer Chamber: The sterilization chamber having total internal volume is 20 m3 & max. usable volume
15.8 m3 which have double walled, corrosion and gas resistant and is constructed of stainless steel and is
designed to withstand the extreme pressures and elevated temperatures utilized in the EO sterilization
process. The inner surface of sterilizer is smoothly finished to minimize gas deposits. The chamber is
insulated against heat emission and the jacket is connected to the warm water circulation arrangement.
The sterilizer doors have a quick release locking arrangement with door opening. Suitable safety locking
arrangement has provided for the door.
The sterilizer has been provided with an automatic programmable panel with memory for preset operating
sequence of all programs of operation. Review the temperature of each channel. T1 is selected as a master
channel to control the cycle.
Independent system such as pressure gauge, temperature gauge, has been provided with the sterilizer for
proper operation and monitoring of sterilizing process to cross check between control and monitoring which
identifies any discrepancies and indicates fault so that an ineffective process doesn’t appears effective.
The ETO gas sterilizer is fully automatic controlled by computer. The control system layout for the PC based
system for ETO sterilizer is as under:
Main
Anal Relay Powe Temperat
mother Digital Control
og Outp r ure
board I/O Panel
Inpu ut Suppl Transmitt
with Intel Cards
t Cards y er Cards
Pentium
Card Card
Processor
s s
CPU & Control Box
System Monitor Normal computer

key board
Visual user interface
Operator interface

5 of 34
Revision No : 0.0
<<<<X<

STRATEGY FOR PERFORMANCE QUALIFICATION
 Loading configuration of 3 way stop cock is used for performance qualification of EO

sterilizer (ETO-04), throughout performance qualification, we are taking 10 no of cycle. 9 cycles are for PPQ
with parallel MPQ. & 10th cycle is additional cycle for PPQ. As a PCD (as an internal PCD), we are choosing
the 3 way stop cock with extension tube. The location of BI’s shall be selected where temperature was
found less in empty cycle study (i.e. cold point) during OQ run.
 If the fresh (Non-sterile) load is not available then we will take sterile load for fractional &
half cycle to performed the validation & testing will be performed after validation cycle (fractional & half
cycle)
 For Physical performance qualification testing, product packaging & functional testing will
performed.
 For microbiological performance qualification testing product bio-burden, Biological

Indicator test & product sterility test, LAL test, EO residual test would be performed.
 Biological indicators (BI) shall be placed inside the articles as defined in diagram-1.0,
product for sterility testing, BET test & product functioning testing shall be placed at different locations in
sterilization chamber.
 Each new validation cycle shall be taken after getting the results of biological indicators of
previous validation cycles.
Sterilization load configuration
Selection of Load used for Performance qualification (PQ)
The 3 way stop cock product load used for PQ is representative of that to be sterilized routinely/ most
frequently and shall be defined based upon the most challenging routine load among the family. Product
three way stop cock which have 288 corrugated boxes during sterilization load for validation of EO
sterilization process. Loading pattern for three way stop cock has maximum density & occupies max. volume
of sterilizer among the family which is sterile in EO sterilizer (ETO-04) as mention in Table -1. Factors, which
may be considered in this load, are those likely to affect penetration of heat, humidity and gas.
Table – 1
Maximu Max. Overa

Maximum Densi
Name of m load Sterilize ll
S. Product quantity ty
Product or r densit
No. to be sterilize in (kg/
Corrugat usable y
one cycle m3)
ed Box in volume (kg/
/C-box
Nos. m3) of
load
Three way 139.71 117.14
1 288 Nos. 2,88,000 Nos. 15.8 m3
stop cock kg/m3 kg/m3
.
Three way
112.32 87.07
2 stop cock with 120 Nos. 90,000 Nos. 15.55 m3
extension tube kg/m3 kg/m3
.
IV Cannula
101.29 76.55
3 with three 150 Nos. 1,20,000 nos. 15.48 m3
kg/m3 kg/m3
. way stop

6 of 34
Revision No : 0.0
<<<<X<

cock

7 of 34
Revision No : 0.0
<<<<X<

Maximu Max. Overa

Maximum Densi
Name of m load Sterili ll
S. Product quantity ty
Product or zer densi
No. to be sterilize in (kg/
Corrugat usabl ty
one cycle m3)
ed e (kg/
Box in volum /C-box
m 3)
Nos. e of load
A.V. Fistula 131.59 106.89
4 204 Nos. 51,000 Nos. 14.71 m3
Needle kg/m3 kg/m3
.
IV. Flow 128.30 99.45
5 120 Nos. 54,000 Nos. 15.55 m3
regulator kg/m3 kg/m3
.
94.81 73.50
6 Polyspike 138 nos. 1,38,000 nos. 15.32 m3
kg/m3 kg/m3
.
113.92 88.31
7 Polysyte 120 Nos. 1,20,000 Nos. 15.55 m3
kg/m3 kg/m3
.
Product Load Configuration
Three way sop cock as a Product load used for PQ should be at least as difficult to sterilize as the most
challenging load. For validation cycles, the maximum quantity of product representing the densest load
configuration that will be processed in full routine cycles must be loaded in the EO sterilizer chamber.
Product Three-way stop cock
Unit packing One Three-way stop cock
Multi unit Duplex box containing 50 blister

packing
Transit Containing 20 multi unit container
container
Actual load A total number of 288 transit container in sterilizer chamber per cycle
The transit containers are placed inside the chamber as per the loading pattern presented in Diagram 1.1.

8 of 34
Revision No : 0.0
<<<<X<

Selection of family representative
A range of product is to be sterilized together in one cycle. The load used for sterilization process
qualification reflects the most difficult combination based on the product design of different sizes, length or
thickness, design complexity & amount and type of packaging as mention below:
 Physical comparison:
Material of Material of Quantity packed in

S. Name of Product
unit transit one Carton
No.
packaging packaging
Three way stop Virgin craft &
1 Paper & PP+PE 500 pcs.
cock with extension Semi craft paper
Film
tube
Paper & Virgin craft &
2 Three way stop 1000 pcs
Tyvek/PVC Semi
cock Film craft paper
IV Cannula with Paper & Virgin craft &
3 500 / 400 pcs.
three way stop Tyvek/PVC Film Semi craft paper
cock
Virgin craft &
4 Infusion (IV) Set Paper & PE Film 500 / 400 pcs.
Semi craft paper
Virgin craft &
5 A.V. Fistula Needle Paper & PE Film 25 set of blister
Semi
. craft paper
Virgin craft &
6 Polyspike Paper & PE Film 1000 pcs.
Semi craft paper
.
Virgin craft &
7 Polysyte Paper & PE Film 500 & 1000 pcs.
Semi craft paper
.
 Dimensional comparison:
S Dimension of Dimension of Dimension of transit
Name of Product
. unit packaging Duplex packaging
N
o
.
Three way stop
1 220 x 70 mm 250 x 230 x 165 850 x 470 x 270 mm
cock with
mm
extension tube
2 Three way stop cock 38 x 75 mm 145 x 135 x 105 550 x 280 x 310 mm
mm
IV Cannula with
3 133 x 50mm 170 x 135 x 50 695 x 325 x 185 mm
three way stop
mm
cock
4 Infusion (IV) Set 100 x 235 mm N/A 590 x 470 x 400 mm
5 A.V. Fistula Needle 450 x 65 mm 500 x 110 x 140 575 x 525 x 305 mm
. mm
6 Polyspike 105 x 55 mm 215 x 205 x 160 850 x 450 x 225 mm
. mm

9 of 34
Revision No : 0.0
<<<<X<

7 Polysyte 220 x 70 mm 250 x 230 x 165 840 x 460 x 260 mm

. mm

10 of 34
Revision No : 0.0
<<<<X<
PRE-REQUISITE FOR VALIDATION
The following requisite must be fulfilled before the ethylene oxide sterilization process validation.
Certification of EO gas
Ethylene oxide gas use as sterilant in the ratio of EO gas (30 %) + CO 2 (70 %). Concentration of EO gas
shall be verified by supplier’s certificate.
Calibration
All critical measuring devices e.g. temperature sensors, humidity sensor, pressure gauges, utilized on
ethylene oxide sterilizers which may impact the quality of the process are calibrated to traceable national or
international standards.
Assessment of product Bio-burden
Population of viable microorganisms on finished product shall be determined before the sterilization process.
It is collected to quantify the amount of product contamination. At least 5 unsterilized samples from the
representative EO sterilized product families to be tested for bio-burden. The product bio burden is tested as
per GTP/QC/12 and record is maintained in F/QA /40.
Product Bio- Burden is tested as per ISO 11737-1. To ensure the product with low bio- burden the product is
manufactured in class 7 at rest and class 8 in dynamic condition.
Biological Indicator (Spore Strips-106)
For the validation study, biological indicators (BIs) of Bacillus Atrophaeous with a minimum population of
spore of 106 are used to challenge EO cycles. The population label claim from each lot of biological
indicators used for validation testing must be verified as per in-house specification. A supplier certificate can
be supported to verify the population of biological indicator count.

11 of 34
Revision No : 0.0
<<<<X<

Requirement for BI’s/ Temperature sensors/ Humidity sensor for validation
A minimum of 40 biological indicators (Spore strips/ or ampoules), 20 temperature sensors and 8 humidity
sensors are to be placed in one validation cycle (Table –2). Highlighted areas are the locations of spore
strips (BIs) as per the Diagram - 1.1. Temperature sensors and humidity sensors kept inside the corrugated
box. Except from these, two temperature sensors & two humidity sensors are placed in to the sterilizer
chamber for monitoring. One temperature sensor (T-3) is kept inside the gas supply line to record the EO
gas inlet temperature.
Table-2:
Minimum no. of BI’s / Temperature sensors/ Humidity
Sterilizer Size of sensors to be used for PQ
No. sterilize
BI’s Temperature Humidity
r
sensor sensor
ETO-04 20 m3 40 Nos. 20 8
Selection of Process challenging device (PCD)
A process challenge device is a device used to assess the effective performance of a sterilization process by
providing a challenge to the process that is equal to or greater than the challenge posed by the most
difficult item routinely processed.
The selection of process challenging device is based on product design which is influence the sterilization
process. The three way stop cock with extension tube is designed in such a way that product is fully closed
from all side of device & tubing product with longest lumen, having most convoluted passageways on basis
of design. Average Product bio-burden is higher than the other product from the family. We compared to
various product into the family at various aspect for selection of PCD as mention in table -3 & 4 which
deemed difficult to sterile product among the family & will be used as the BI’s (Bacillus atropheous) carrier.
Table-3
Overall
Average
S Name of density Unit
Raw material used product
. Product (kg/m3) packaging
Bio-
N of load material
burden
o used
.
Three way stop HDPE, Paper &
1 117.14 5 CFU/ Device
cock Polycarbonate, Tyvek/PVC Film
kg/m3
Polypropylene
Three way stop
HDPE,
2 cock with 87.07 kg/m3 Paper & PE Film 10 CFU/
Polycarbonate,
extension Device
Polypropylene, ABS
tube
IV Cannula HDPE, Polycarbonate,
Paper &
3 with 76.55 kg/m3 Polypropylene, 6 CFU/ Device
Tyvek/PVC Film
three way Poly Vinyl
12 of 34
Revision No : 0.0
<<<<X<

stop cock Chloride

13 of 34
Revision No : 0.0
<<<<X<

Overall
Average
S Name of density Unit
Raw material used product
. Product (kg/m3) packaging
Bio-
N of material
burden
o load used
.
A.V. Fistula Poly Vinyl
4 106.89 Paper & PE 7 CFU/
Needle Chloride, LDPE, Film
kg/m3 Device
HDPE, ABS
Polycarbonate,
Rigid Polyvinyl
5 IV. Flow 99.45 kg/m3 Paper & PE 7 CFU/
chloride, TPR, Rigid Film
regulator Device
Polyvinyl chloride,
HDPE, ABS
Acrylonitrile
6 Polyspike 73.50 kg/m3 Butadiene Paper & PE 5 CFU/ Device
Styrene (ABS), Film
Polypropylene
Table-4
Internal Max. Size of

S Name Status of Opening Closures
surface Lumen
. of of Product (Less/ More)
area (Product
N Product Length)
compariso
o
n
.
Having a opening
Three way Less Less closure
1 from 4 cm
stop cock internal present
protective cap &
surface
from the thread of
area
luer lock.
Three way Fully closed. EO Gas
More more
2 stop pass only from Luer 200 cm
internal closure
cock with lock & protective
surface area present
extension cap.
tube
IV Cannula Having a opening
Less Less closure
3 with from 12 cm
internal present
three way protective cap &
surface
stop cock from the thread of
area
luer lock.
Having a opening
A.V. Fistula More Less closure
4 from 36 cm
Needle internal present
protective cap &
surface area
from the thread of
luer lock.
Having a opening
IV. Flow More Less closure
5 from protective cap 50 cm
regulator internal present
& from
the thread of luer surface area
lock.

14 of 34
Revision No : 0.0
<<<<X<

Having a opening
Less Less closure
6 Polyspike from cover & from 6 cm
internal present
the thread
surface
of luer lock.
area

15 of 34
Revision No : 0.0
<<<<X<

Placement of Biological Indicator at difficult to sterilize locations:
To create a challenge to the sterilization process, PCD, Bacillus atropheous having minimum 10 6 population,
by placing biological indicators in the product at location where sterilizing condition are most difficult to
achieve. Although the product are designed in such a way that EO gas reaches to all locations in the product
for example the passage from luer cover help EO gas to reach inside the channel &, handle, screw cap,
tubing etc. But still biological indicators are kept at the locations indicated below diagram 1.0.
Spore Strips
Diagram – 1.0

16 of 34
Revision No : 0.0
<<<<X<

Location of Data logger (Sensors), Bi’s & samples for physical &
microbiological testing in to the product load volume
Three way stop cock is choosing as product load for validation. Temperature and humidity sensors should be
located within the sterile barrier system or amongst the unit packages in the sterilization load different
location i.e trolley centers, trolley edges & surfaces. A minimum of 20 numbers of the sensors for
temperature and 8 numbers of sensors for humidity measurement are located at different locations into the
product load volume. Samples for physical testing (like product functional testing & packaging testing) as
well as microbiological testing (like product sterility, BET, EO etc.) are placed different location. Following
are the location for the placement of temperature sensors, Humidity sensors & Biological indicator are given
below:
Trolley Test cases (Corrugated Boxes) Remarks

No.
1 1T, 8, 20H, 23T, 25, 29H, 42T
 Each Test case
2 57T, 59H, 69T, 75, 80, 95T contains 1 Biological
indicator.
3 98T, 111, 113T, 123T, 128T, 130, 143H
 Test cases with
4 145T, 157, 164T, 167, 174H, 190T ‘T’ contains
5 196T, 204H, 211, 213, 220T, 230T, 238 temperature sensor.
 Test cases with like
6 242H, 253T, 260, 266T, 273H, 274T, ‘H’ contains humidity
282T sensor.
Chemical indicator tape is pasted on all containers kept inside the sterilizer chamber. All boxes containing the
biological indicators shall be marked for proper identification.
LOCATION PATTERN FOR PRODUCT
TROLLEY-1 TROLLEY-2
2 15 10 5 68 63 58 53
0
19 14 9 4 67 62 5 52
7
18 13 8 3 66 61 56 51
17 12 7 2 65 60 55 50
16 11 6 1 64 5 54 49
9
28 24 76 72
27 23 75 71
26 22 74 70
25 21 73 69
48 43 38 33 96 91 86 81
47 4 37 32 9 90 85 8
2 5 0
17 of 34
Revision No : 0.0
<<<<X<

46 41 36 31 94 89 84 79
45 40 35 30 93 88 83 78
44 39 34 2 92 87 82 77
9

18 of 34
Revision No : 0.0
<<<<X<

TROLLEY-3 TROLLEY-4
11 1 10 10 1 15 15 14
6 1 6 1 6 9 4 9
1 4
11 11 10 10 16 15 15 14
5 0 5 0 3 8 3 8
11 10 10 99 16 1 15 14
4 9 4 2 5 2 7
7
1 10 10 9 16 15 15 14
1 8 3 8 1 6 1 6
3
11 10 10 97 16 15 15 1
2 7 2 0 5 0 4
5
124 120 172 168

123 119 171 167
122 118 170 166
121 117 169 165
14 13 13 12 19 18 18 17
4 9 4 9 2 7 2 7
1 13 13 1 19 18 18 17
4 8 3 2 1 6 1 6
3 8
14 13 13 12 1 18 18 17
2 7 2 7 9 5 0 5
0
14 13 13 12 18 18 17 1
1 6 1 6 9 4 9 7
4
14 13 1 12 18 18 17 17
0 5 3 5 8 3 8 3
0
TROLLEY-5 TROLLEY-6
21 20 20 19 2 25 25 24
2 7 2 7 6 5 0 5
0
2 20 20 1 25 25 24 24
1 6 1 9 9 4 9 4

19 of 34
Revision No : 0.0
<<<<X<

1 6
21 20 20 19 25 2 24 24
0 5 0 5 8 5 8 3
3
20 2 19 19 25 25 24 2
9 0 9 4 7 2 7 4
4 2
20 20 19 19 25 25 24 24
8 3 8 3 6 1 6 1
220 216 268 264

219 215 267 263
218 214 266 262
217 213 265 261
24 23 2 22 28 28 27 2
0 5 3 5 8 3 8 7
0 3
23 23 22 22 28 2 27 27
9 4 9 4 7 8 7 2
2
2 23 22 22 28 28 27 27
3 3 8 3 6 1 6 1
8
23 23 22 22 28 28 27 27
7 2 7 2 5 0 5 0
23 23 22 22 28 27 2 26
6 1 6 1 4 9 7 9
4
Diagram -1.1

20 of 34
Revision No : 0.0
<<<<X<

LOADING PICTURE FOR PRODUCT - 3 WAY STOP COCK

21 of 34
Revision No : 0.0
<<<<X<
Schematic Diagram Showing Location of RTD Sensors & humidity sensor inside chamber
EO gas pipeline
UNLOADING SIDE
T3
RH 2
T1
T2
RH 1
Steam Inlet point
Steam Inlet point

LOADING SIDE
Location of Sensors:
T2, & RH2 : Left wall of sterilizer from un-sterile side

T 1 & RH 1 : Right wall of sterilizer from un-sterile side
T3 : EO Gas line

22 of 34
Revision No : 0.0
<<<<X<
PERFORMANCE QUALIFICATION – PHYSICAL (PPQ)
Physical PQ (PPQ) shall demonstrate
 That the specified acceptance criteria are met throughout the load for the duration of the proposed
routine process specification, and
 Reproducibility of the process
The PPQ shall include following planned qualification cycles in table-3, in which all the specified acceptance
criteria shall be met. PPQ is performed in parallel with MPQ runs; therefore a minimum of one additional
PPQ run (280 minutes) shall be performed using the full routine process specification.
Table-3
Sr. Cycle no. Exposure Time Cycle Description
No.
1 1st cycle 20 Minutes Fractional cycle
2 2nd cycle 40 Minutes Fractional cycle
3 3rd cycle 60 Minutes Fractional cycle
4 4th cycle 80 Minutes Fractional cycle
5 5th cycle 100 Minutes Fractional cycle
6 6th cycle 120 Minutes Half Cycle
9 9th cycle 280 Minutes Full cycle
10 10th cycle 280 Minutes Full cycle
The EO sterilizer is equipped with computer control software to meet the above requirement. EO Sterilization process
steps are described below:
Minimum Temperature of product to enter the Sterilization Process shall be recorded.

Pre-Humidification Phase: The sterilization load is placed inside the sterilization chamber. The load is
first subjected to humidification for 5 ± 1 minute to attain the required humidity for preconditioning.
Preconditioning: Treatment of product prior to the sterilization cycle in the chamber to attain specified
limits for temperature and relative humidity. The sterilization chamber is used as pre conditioning area. The
load is kept on pallets as per loading pattern.
The load in pre-conditioning is kept for 60 minutes to attain a minimum temperature of 45ºC ± 5ºC and
humidity between 30% to 90% at the end of preconditioning.
Conditioning: Treatment of product within the sterilization cycle, but prior to sterilant admission, to attain
a predetermined temperature and relative humidity. This part of the sterilization process is carried out under
vacuum.
Since the pre-conditioning & conditioning area are same so all parameters like loading pattern and location
of temperature & humidity sensors remain same.
 Initial Vacuum Phase

To safely deliver the EO process, air must be removed from the chamber. The air is removed by pulling a
deep vacuum up to -0.75kg/cm2 to remove the air. The evacuation rate is selected to assure package
integrity is maintained. This will assure that the
23 of 34
Revision No : 0.0
<<<<X<
package’s sterile barrier properties are maintained, thus ultimately protecting the sterility of the product
once the process is complete. The rate of vacuum up-to -0.75kg/cm2 is approximate 30 ± 15 minute.
 Vacuum Hold phase (Depth & Rate of Attainment of Vacuum):

A vacuum to -0.75 kg/cm2 is drawn for evacuation of air inside the chamber for effective sterilization. The
vacuum of sterilizer is kept on hold for 10 ±1 minutes. The vacuum shall not drop during this period. If
there is no leakage the vacuum will remain to the same pressure.
If there is any leakage is detected, the sterilizer is not taken for sterilization unless the problem is solved
and approval given by Maintenance department.
 RH Injection phase & Dwell Phase

If humidification is required, it is added by injecting steam. Steam is utilized to humidify the load and is not
intended to be a sterilant. The consistency of steam supply can be determined by analysis of condensate.
The recording system is provided which will record the pressure & time of injected steam. Usually the
ambient humidity of the product is mostly within the required limit hence injection of steam is not required.
In any case if humidity is less steam can be injected for one time for 2-5 minutes & RH is kept hold for 10
±1 minutes. Chamber pressure after insertion of steam (Relative Humidity) shall be maintained -0.75 to -
0.70 kg/cm2. In case, steam is injected more than 0.05kg/cm 2, then evacuation start for vacuum drop
compensation. Chamber temperature should attain 45±5°C, Temperature of the sterilization load shall meet
35±5°C, Humidity of the sterilization load at the end of conditioning load shall be in the range of 30-90 %.
Gas Injection phase (Gas charging)
After conditioning time (RH dwell phase), the gas injection is started. The gas, which is admitted inside the
chamber, is preheated and is in vapor form with a temperature between 20ºC~60ºC. If the temperature
comes below 20ºC, the injection of EO will be automatically stopped.
The amount of gas added was calculated to a particular weight and weighed using a calibrated balance.
However, the evaluation of the EO gas concentration injected can be correlated with the pressure of
sterilizer chamber.
Correlation between computer system and manual system
Parameter Computer controlled Independent manual

gauge
Pressure rise &
0.25 kg/ cm2 ± 0.05 kg/ cm2 0.25 kg/ cm2 ± 0.05 kg/ cm2
Terminal Pressure
Chamber 45ºC ± 5ºC 45ºC ± 5ºC
Temperature
Since between preconditioning / conditioning are performed in sterilizer chamber, therefore maximum
elapsed time between condition and sterilization is NIL.
The calculation for EO concentration mg/lit is taken as below -
Wt. of EO gas X Mixture of EO in Gas (30%) X 10000 EO concentration (mg/liter) = ----------------------------

---------------------------------------
Area of Sterilizer in Liters (20000)

24 of 34
Revision No : 0.0
<<<<X<

Exposure phase: After the gas has been injected, the exposure phase of the process is performed. This is
the phase in which the product is exposed to heat, relative humidity, and gas for a predetermined amount of
time. The amount of exposure time is determined after careful analysis of the product, load configuration,
and desired level of sterility. During EO exposure period, sterilizer chamber temperature should have
45±5°C, temperature of sterilization load should be 35±5°C.
Aeration
After completion of required exposure time, Ethylene Oxide gas is removed/ flushed from the chamber and
package (product) up to -0.75 kg/cm² with the rate approximate 40±20 minute and the Aeration is started.
Flushing is done by removing the air from chamber and package by creating a vacuum deep to -0.65
kg/cm². The rate of air wash phase is approximate 30±15 minutes. Air flushing is done two times. During
aeration temperature within the chamber/ area (should be 35°C - 60°C) & Temperature of the sterilization
load (30°C – 50 °C) shall be recorded.
ACCEPTANCE CRITERIA FOR PERFORMANCE QUALIFICATION- PPQ
The parameters for standard process used for routine sterilization cycle, which is being validated are as
follows:
Acceptance
Sr. CYCLE PHASE Specificati
Criteria/ Limit
No on
.
Minimum Temperature of product to enter the Sterilization
a) NLT 20 °C NLT 20 °C
Process
Pre-Humidification Phase (Steam Injection)
1
Time 5 minutes  1 minute
b) Preconditioning (if used)
1 Time in Chamber/Area 60 minutes  2 minute
2 Temperature of Chamber/Area 45°C  5°C
3 Humidity of Chamber/Area 60% 30% ~ 90%
4 Temperature of sterilization load 35°C  5C
5 Humidity of sterilization load 60% 30% ~ 90%
c) Vacuum levels and rate of evacuation (if used)
1 Vacuum levels -0.75  0.01Kg/cm2
Kg/cm2
2 Vacuum Rate 30 minutes  15 minutes
3 Holding time under vacuum 10 minutes ± 1 minutes
e) Conditioning (RH Dwell Phase)
Chamber Pressure after insertion of steam (Relative NLT -0.70
1 kg/cm2 NLT -0.70 kg/cm2
Humidity)
One time &
2 Number of steam pulses & Duration 2-5 minutes
2
minutes
3 Conditioning time (RH Dwell Time) 10 minutes ± 1 minutes

25 of 34
Revision No : 0.0
<<<<X<

4 Chamber Temperature 45°C  5C

26 of 34
Revision No : 0.0
<<<<X<

Acceptance
Sr. CYCLE PHASE Specificati
Criteria/ Limit
No on
.
Temperature of the sterilization load at the end of
5 conditioning 35 °C  5C
6 Humidity of the sterilization load at the end of conditioning 60% 30% ~ 90%
f) EO Injection and Exposure
1 EO injection pressure rise (ΔP) 0.25 Kg/cm2 ± 0.05kg/cm2
2 EO injection time 40 min  20 minute
3 Terminal pressure of EO injection phase 0.25 Kg/cm 2
± 0.05 Kg/cm2
4 EO gas Inlet Temperature NLT 20C 20C ~ 60C
5 Mass of EO used 36.7 kg 35 kg ~ 38.3 kg
6 Concentration of EO 550 mg/lt ± 25mg/lit
7 Sterilizer chamber temperature 45°C  5 C
8 Exposure time As point ± 1 minutes
no. 6.0
9 Temperature of the sterilization load 35°C  5 C
h) Aeration
-0.75  0.01 kg/cm2 @ 
1. Gas Discharge phase
kg/cm2 @ 20 minutes
40 Minutes.
2. Air Inlet phase Up to 0 ± 0.05kg/cm2
kg/cm2
-0.65  0.01 kg/cm2 @ 
3. Air washing phase and Rate
kg/cm2 @ 15 minutes
30 Minutes.
4. Air inlet phase Up to 0 ± 0.05kg/cm2
kg/cm2
5. Temperature within the Chamber/Area 45°C ± 15 C
6. Temperature of the sterilization load 40°C ± 10 C
7. Total Aeration 2 Times 2 Times
PERFORMANCE QUALIFICATION– MICROBIOLOGICAL (MPQ)
The total inactivation of microorganisms using EO is attained when sterilizing conditions are met within the
chamber. The four active ingredients required to deliver a successful process are:
• Heat/Temperature
• Moisture
• Gas concentration
• Exposure Time
The microbiological Performance Qualification (MPQ) shall demonstrate that, on application of the
sterilization process, the specified requirements for sterility are met. Exposure time is the key parameter that
is varied during microbiological qualification to provide assurance that MPQ delivers less lethality than the
normal production process. Determination of lethal rate of sterilization process (Bi/ bioburden approach) is
carried out by fraction negative method {Annex A, clause A.1.3. (b)} & conservative determination of lethal

27 of 34
Revision No : 0.0
<<<<X<

rate of the sterilization process by Over Kill Approach {clause B.1.2 (a) of EN ISO 11135-1:2007/ ISO
11135:2014}:
The Overkill Approach is most commonly used to validate ethylene oxide cycles used in the sterilization of
medical devices. The overkill method is based on demonstrating that the sterilization of a microbial

28 of 34
Revision No : 0.0
<<<<X<

challenge/ biological indicator (Bacillus atropheous) exceeds the challenge posed by the bio-burden of the
product.
MPQ shall include at least three half cycles for determination of the minimum time of exposure to EO, with
all other process parameters except time remaining constant, at which biological indicators are not showing
any growth. The specified exposure time should be at least double this minimum time.
Following cycles to be taken during the process qualification & calculation of D- Value by Limited Holcomb-
Spearman-Karber Procedure (LHSKP)
Table-2 — calculation of D- Value by LHSKP with constant time intervals and constant number
of samples
Sr. Number of test

No. Cycle Exposure Nature of
samples
Description Time Growth
exposed
1 1st cycle 40 BI’s 20 Minutes Complete Growth
2 2nd cycle 40 BI’s 40 Minutes Partial Growth
3 3rd cycle 40 BI’s 60 Minutes Partial Growth
4 4th cycle 40 BI’s 80 Minutes Partial Growth
5 5th cycle 40 BI’s 100 Minutes Partial Growth
6 6th cycle 40 BI’s 120 Minutes No Growth
10 10th cycle NA 280 Minutes NA
D value calculation: Formula to be used for fraction negative method for D value calculation as per clause
C.3.2 Limited Holcomb-Spearman-Karber Procedure (LHSKP) of ISO 14161:2009 (E).
After exposure, the test samples are inoculated into Soyabean Casein Digest Medium and observe the
growth for 7 days.
ACCEPTANCE CRITERIA FOR PERFORMANCE QUALIFICATION-MICROBIOLOGICAL
Testing for product bio-burden
At least 5 unsterilized samples from the representative EO sterilized product families to be tested for bio-
burden. The product bio burden is tested as per ISO 11737-1 & GTP/QC/12 and record is maintained in
F/QA /40. Product Bio- Burden should be NMT 100 CFU/device.
Testing of biological indicator
 Exposed biological indicator (spore strips) in 40 nos. for each cycle are
inoculated in Soyabean Casein Digest Medium Previously sterilized at 121ºC for 15 minutes and kept at
30-35ºC in BOD incubator for 7 days. . The media is monitored daily for any growth of microorganism.
 Exposed biological indicator (ampoules) (if used) are crush & incubate processed
and control BI’S for 48 hours at 30-35ºC in BOD incubator.
The result shall comply test plan as per clause 7.0 and record shall be maintained F/QA/35. BI’s should be
incubated within 24 hours after cycle completion (unloading).

29 of 34
Revision No : 0.0
<<<<X<
Ethylene Oxide Residual Testing
The sample (minimum 5 in nos.) would be drawn out for EO residual testing as mention below:
 For fractional cycle- Sample would be collected after execution of full routine cycle
 For half cycle- Sample would be collected after execution of full routine cycle.
 For Full cycle – Sample would be collected.
This test is used to determine the concentration of Ethylene Oxide in compliance with the EN ISO 10993-
7:2008 standard. EO residual testing will be performed by external NABL certified Lab. Acceptable residual
limits for Ethylene oxide is NMT 4mg/ 24 hrs, & for Ethylene Chlorohydrin is NMT 9 mg/ 24 hrs. Simulative
used extraction method is using for testing of EO residual.
Note- if the fresh load is not available then we will take sterile load for fractional & half cycle & performed
the EO residual test after the fractional & half cycle execution.
LAL testing (Bacterial Endotoxin Testing)
Take 3 sterilized samples from the EO sterilized product to be tested for bacterial endotoxin test. The
sterilized product shall comply the Bacterial Endotoxin Testing using Gel-clot method as per GTP/QC/10.
Product sterility Testing
20 sterilized samples from the EO sterilized product load to be tested for sterility test. The sterilized
product shall comply the Sterility test as per GTP/QC/09, revision no.-02.
OTHER QA EVALUATION TEST
Product Package Testing
Product is tested randomly for the effect of sterilization on packaging condition, printing matter, sealing or
any other effect due to temperature & humidity as well as functioning of product. The effect of sterilization
shall not have any detrimental effect on the package.
Product Functional Testing

Product shall be tested for functional testing for effect of sterilization processes during finish product testing.
The effect of sterilization shall not have any detrimental effect on the functioning of product.
9.0 COLLECTION OF DATA/ SAMPLES
Data logger, internal PCD & samples shall be collected after the completion of cycle for further evaluation.
For process parameter analysis, collect the data from the routine graph as well as data logger cycle wise.
10.0 REVIEW OF DATA
All the data will be carefully reviewed & should be met as per specification/ acceptance criteria.

30 of 34
Revision No : 0.0
<<<<X<
11.0 DEVIATION REPORT
Any deviation from the acceptance criteria of the specific check point shall be reported in document
F/QA/168 Revision no. - 00 and decision should be taken for the rejection, replacement or rectification of the
equipment/component.
12.0 DISPOSITION OF PRODUCT
Fractional cycle & half cycle are followed by Full sterilization cycle during PPQ & MPQ. So product will be
verified product packaging, functioning test & EO residual test after repeatability sterilization process. If the
product failed to achieve the acceptance criteria in any test like product functioning test, EO residual test &
Product packaging test, then it should be disposed off or discarded in presence of QA & recorded in
validation report.
DATA RECORDING
Data shall be recording in following annexure & format during performance qualification.
 Product bio-burden shall be recorded in format – F/QA/40.
 Testing record for biological indicators shall be recorded in format - F/QA/35.
 Sterility testing record shall be recorded in format - F/QA/09.
 BET testing record shall be recorded in format - F/QA/49.
 Product functional testing shall be recorded in format – F/QA/52.
 For report for effect of EO sterilization on product package shall be recorded in Annexure-I
Annexure-I
PRODUCT : DOCUMENT NO. -
CYCLE NO. : DATE :
EFFECT ON EFFECT
MODE PACKING
OF
OF MATERIA
SEALIN PRINTI DEFORMATIO COLOR HUMIDI
PACKIN L
G NG N TY ON
G
PAPER
Medical
UNIT grade
PACKING paper /
Tyvek &
rigid
PVC
film
DUPLEX Card
CARTON board
paper
31 of 34
Revision No : 0.0
CORRUGA Semi
TED BOX craft
paper

32 of 34
Revision No : 0.0
<<<<X<
14.0 IDENTIFICATION OF PERSONNEL PERFORMING VALIDATION OF EO STERILIZATION PROCESS

Name of the identified persons performing the validation of EO sterilization process are as under:
TRAINING GIVEN BY
Name
Designation
Signature/ Date
TRAINING TO EXECUTION TEAM
Name Designation Department Signature Date
Venue
Date of training
Duration of training
15.0 EVALUATION
The EO sterilizer shall be tested for its killing efficiency by review of the data for biological indicator and RTD
sensors on different locations.
Chamber leak test of EO sterilizer shall be performed (Integrated in Standard process test) and pressure
drop during vacuum hold time shall be observed.
The RTD sensors and biological indicators shall be placed in to the product load volume at different
positions. At any time interval of sterilization hold time no RTD sensor shall show the temperature difference
± 5°C from the mean temperature of all the temperatures. The chemical indicators shall show the colour
changes as per the instructions of the manufacturer.

33 of 34
Revision No : 0.0
<<<<X<
16.0 VALIDATION REPORT
The validation report shall include but not limited to - date of study initiated, date completed, name and
signature of execution team, material and instruments used, observations made, problems encountered,
completeness of information collected, summary of deviation report, results of tests (conclusion), do results
meet acceptance criteria, other information relevant to the study and conclusions on the validity of the
equipment/system.
REQUALIFICATION CRITERIA
EO sterilizer to be re-qualified on: -
 Once in a year
 Major sterilizer repairs and changes
 Changes to construction or relocation
 Unexplained sterility failures in routine sterilization
 Changes to product or packaging
 Modification to the sterilizing agent

34 of 34

PQ Protocol - Final

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

PQ Protocol - Final

Uploaded by

Copyright:

Available Formats

Medivac Surgical Pvt. Ltd.-Kutch, Gujarat India. Doc No : MVS/QP/19.

1. APPROVAL OF PERFORMANCE QUALIFICATION PROTOCOL

DEPARTMENT NAME DESIGNATI SIGNATURE DATE

DEPARTME NAME DESIGNATI SIGNATURE DATE

Plant & Aadish

DEPARTMENT NAME DESIGNATION SIGNATURE DATE

Page No Prepared By Reviewed By Approved By

The scope of this validation is as under for Sterilization of

The detail of equipment is as under:

Name of the Equipment : EO (Ethylene Oxide) Sterilizer- 01

Technical Identification No. : MSPL/ETO/01

Model No. : HDX-EN-20

Inner container volume : 20 m3

Equipment’s Location : Ground floor near Finished Goods.

DEPARTMENT DESIGNATION RESPONSIBILITY

Shall prepare validation Protocol & report as

Shall approve the validation protocol &

Page No Prepared By Reviewed By Approved By

5. MATERIALS, INSTRUMENTS AND DOCUMENTS

3. Validation procedure (QP/QAD/25, Rev. No.-09)

6. Chemical indicator Tape

7. Ethylene oxide gas cylinder (90% EO & 10% CO2)

8. Calibrated Weighing balance

9. Bio burden on product (GTP/QC/12)

10. Population verification of Biological indicators

6. REFERENCE INTERNATIONAL STANDARDS USED FOR VALIDATION

 BS EN ISO 11737-1:2006, Sterilization of medical devices - Microbiological methods -Part 1:

Page No Prepared By Reviewed By Approved By

7. SYSTEM/ EQUIPMENT DESCRIPTION - Brief description of equipment (sterilizer)

The sterilizer (ETO-01) is having inner dimension of chamber as under:

Length: 9.0 meter

System Monitor Normal computer

Page No Prepared By Reviewed By Approved By

Effective Date : 01-06-2018

STRATEGY FOR PERFORMANCE QUALIFICATION

 Loading configuration of 3 way stop cock is used for performance qualification of EO

 For microbiological performance qualification testing product bio-burden, Biological

Sterilization load configuration

Selection of Load used for Performance qualification (PQ)

Maximu Max. Overa

Page No Prepared By Reviewed By Approved By

Effective Date : 01-06-2018

Page No Prepared By Reviewed By Approved By

Effective Date : 01-06-2018

Maximu Max. Overa

Product Load Configuration

Product Three-way stop cock

Unit packing One Three-way stop cock

Multi unit Duplex box containing 50 blister

Page No Prepared By Reviewed By Approved By

Effective Date : 01-06-2018

Selection of family representative

Material of Material of Quantity packed in

Page No Prepared By Reviewed By Approved By

Effective Date : 01-06-2018

7 Polysyte 220 x 70 mm 250 x 230 x 165 840 x 460 x 260 mm

Page No Prepared By Reviewed By Approved By

PRE-REQUISITE FOR VALIDATION

Assessment of product Bio-burden

Biological Indicator (Spore Strips-106)

Page No Prepared By Reviewed By Approved By

Effective Date : 01-06-2018

Requirement for BI’s/ Temperature sensors/ Humidity sensor for validation