Surge Laboratories (Pvt.) Ltd.

ASEPTIC PROCESS VALIDATION

PROTOCOL (Dry Powder
Injectable Area)

Quality Assurance Department

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 1 of 13

There are 12 numbers of pages in this Protocol.

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Surge Laboratories (Pvt.) Ltd.

ASEPTIC PROCESS VALIDATION
PROTOCOL (Dry Powder
Injectable Area)

Quality Assurance Department

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 2 of 13

We have reviewed the Aseptic Process Validation Protocol Micro-10-QA-002 for the Dry
Powder Injectable and found it acceptable.

Prepared By: ____________________________

Date: _______________

Umair Zubair (Microbiologist)

Reviewed By: ____________________________

Date: _______________

Kamran Atif (Manager QA)

Approved By: ____________________________

Date: _______________

Asad-Uz-Zaman (GM P & P)

Approved By: ____________________________

Date: _______________

Mutasma Yasmeen (Head of Technical Operations)

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Surge Laboratories (Pvt.) Ltd.

Quality Assurance Department

ASEPTIC PROCESS VALIDATION

PROTOCOL (Dry Powder
Injectable Area)

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 3 of 13

TABLE OF CONTENTS
S. No.

TITLE

01

Objective

02

Unit Operation

03

Acceptance Criteria

04

Reference

05

Process Flow

06

Equipment

07

Material

08

Procedure

09

Incubation & Inspection

10

Deviation & Change control

11

Analysis of Data

12

Re-Validation

13

Conclusion & Issuance of Certificate

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Surge Laboratories (Pvt.) Ltd.

ASEPTIC PROCESS VALIDATION
PROTOCOL (Dry Powder
Injectable Area)

Quality Assurance Department

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 4 of 13

HISTORY SUMMARY SHEET

S. No

Nature of Change

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Page/Line

Sign.

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Surge Laboratories (Pvt.) Ltd.

ASEPTIC PROCESS VALIDATION
PROTOCOL (Dry Powder
Injectable Area)

Quality Assurance Department

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 5 of 13

1.0 OBJECTIVE:
1.1 To validate that Aseptic Filling of Dry Powder Injectable meet all pre-established
cGMP requirements to conform as Sterile Product and to demonstrate that online filling
operations of sterile Polyethylene Glycol 8000 (PEG 8000) Powder & Tryptic Soya Broth
Media conform to the actual filling process.

2.0 UNIT OF OPERATION:

2.1 Online Simulated media fill trial with Sterile Polyethylene Glycol 8000 (PEG) and
Tryptic Soya Broth (TSB) in Vial Filling Area.

3.0 ACCEPTANCE CRITERIA:

3.1 Minimum three thousand vials or units of 250mg will be filled with sterile
Polyethylene Glycol 8000 (PEG 8000) powder & Tryptic Soya Broth Media
3.2 Minimum three thousand vials or units of 1000mg will be filled with sterile
Polyethylene Glycol 8000 (PEG 8000) powder & Tryptic Soya Broth Media
3.3 Less than or equal to 0.1% contamination rate is acceptable . Not more than six vials
should be observed positive from the both filling processes out of 6000 or Not more than
three vials from each three thousands (3000) vials of 250mg & 1000mg.

4.0 REFERENCE:
Validation of Aseptic Processes (PI 007-5 1 July 2009), Section 4.3.1.

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Surge Laboratories (Pvt.) Ltd.

Quality Assurance Department

ASEPTIC PROCESS VALIDATION

PROTOCOL (Dry Powder
Injectable Area)

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 6 of 13

5.0 PROCESS FLOW:

Issuance of TSB Media
(0.954 Kg)

Preparation of media.
(31.8L) in WFI & Media in Vessel
(Capacity 50L)

Issuance of 6100 Vials of

both 250mg and 1000mg
capacity

2000 vials
for 500mg

Washing & Sterilization of

Vials.
o
Autoclavation at 250 C for 2
hrs

Sterilization of media at
121oC for 15 min

1950
Sterilization of Filling Accessories
e.g Machine Parts, Uniforms,
Rubber Stopper S.Gloves &
Dusters at 121oC for 1hr

vials for 1000mg

Issuance of PEG 8000 Powder

sterilized by gamma radiation
( Approximately 4.4Kg)

Assembling of Machine Parts

Sterile Process Simulation

Filling of Powder PEG 8000 as fill weight with TSB media (5.3ml per vial)
Sealing of vials
Expose the Settle plates during filling
IPQA check weight & Process after every 45min
Quantity: 3000 vials for 250mg & 3000 vials for 1000mg
Fill Weight: 250mg (225 - 288mg) & 1000mg (900 1150mg)

Transfer of Filled Vials to QC for

Incubation and Visual check for
any turbidity

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Surge Laboratories (Pvt.) Ltd.

ASEPTIC PROCESS VALIDATION
PROTOCOL (Dry Powder
Injectable Area)

Quality Assurance Department

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 7 of 13

6.0 EQUIPMENT:
List of Major Equipment is as below.
Equipment
Name

Vial Washing
Machine

04-VW-073

Dry Oven

04-OV-04

P.V. Sterilizer

04-PVS-51

Vial Area

PQ-04-034

04-VF-55

Vial Filling
Area

PQ-04-028

04-VS-58

Vial Sealing
Area

PQ-04-005

4
5

Equipment
Code

Automatic
Microdoze Vial
Filling Machine
Vial Sealing
Machine

Equipment
Location

Equipment
Operated
Qualification
By
Document #

S. #.

Vial
Washing
Area
Vial
Washing
Area

Checked
By

Verified
By

PQ-04-007
PQ-04-030

7.0 MATERIAL:

Media: Tryptic Soya Broth ~ 0.954 Kg (30g per Liter)

PEG 8000 ~ 4.4Kg
Glass Vials:
3050 Glass Vial 8 ml for 250mg filling operation.
3050 Glass Vial 15ml for 1000mg filling operation.

8.0 PROCEDURE:
Reference: BMO-04-006 & SMP-04-006
8.1 ISSUANCE OF BULK:
Get issue of sealed sterile PEG 8000 powder (Sterilized by Gamma Radiation) from
Material Management & non-sterile TSB media from QC a day in advance as per SOP-

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Surge Laboratories (Pvt.) Ltd.

ASEPTIC PROCESS VALIDATION
PROTOCOL (Dry Powder
Injectable Area)

Quality Assurance Department

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 8 of 13

08-QC-002 and then sterilize it at 121oC for 15 minutes in Steam Sterilizer (Autoclave)
of Dry Powder Injectable Area.
Separately perform sterility test of sterile PEG 8000 powder as per membrane filtration
method as per SOP-08-QC-014. Bioburden of Non sterilized Broth is to be evaluated
beside ve and Positive (+ve) test of Broth for growth formation.
8.2 WASHING & STERILIZATION OF VIALS:
Wash glass vials, with Deionized Water, Filtered Distilled Water, Compressed Air (0.45 &
0.2 Microns) as per SOP-04-005 & SOP-04-008 & sterilize the glass vials as per SOP-04009. Determine the bioburden of washed vials before sterilization as per SOP-08-QC106. Check the sterility of vials after sterilization as per SOP-08-QC-100. Monitor hot air
oven by using Spore Strip bioindicator as per SOP-08-QC-102.
8.3 WASHING & STERILIZATION OF MACHINE PARTS:
Wash & Sterilize the Machine Parts as per SOP-04-009. Monitor autoclave by using
Ampoule bioindicator as per SOP-08-QC-101.
8.4 STERILIZATION OF FILLING ACCESSORIES:
Sterilize the rubber plugs, uniforms, gloves & dusters as per SOP-04-009. Check the
sterility of rubber plugs after sterilization as per SOP-08-QC-100.
8.5 STERILIZATION OF MEDIA:
Prepare the Broth as per SOP-08-QC-002, i.e; 31.8L for 6000 units/vials (5.3 ml in each
vial) in SS container & sterilize it at 121oC for 15 min in Steam Sterilizer (Autoclave) of
Dry Powder Injectable Area. Perform Growth Promotion test of TSB medium as per
SOP-08-QC-027.
8.6 STERILE PROCESS SIMULATION:
8.6.1

All activities are performed under the direct supervision of Qualified Person from
QA, Production & Microbiology Section.

8.6.2

Perform all the activities in the specific classified areas.

8.6.3

Prepare the aseptic area as per SOP-04-001.

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Surge Laboratories (Pvt.) Ltd.

ASEPTIC PROCESS VALIDATION
PROTOCOL (Dry Powder
Injectable Area)

Quality Assurance Department

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 9 of 13

8.6.4

Perform monitoring of Air Changes, Air Velocity of Laminar Flow Hood (LFH) &
Pressure Differential as per SOP-08-QC-058 and air flow pattern as per SOP-08QC-104.

8.6.5

First adjust Powder Filling machine for 250 mg (225 - 288mg) weight & fill
3000vials with(PEG 8000) powder and then immediately fill TSB Broth 5.3 ml in
each vial through Automatic Media Filling Nozzle fitted with compounding vessel
as per SOP-04-012. After Both filling, plug & seal the container.

8.6.6

Perform the filling of 1g on the same day after cleaning & mopping of area with
stay of 30 minutes in the presence of Microbiologist & QA. Now adjust the
machine for 1g (900 1150mg) weight to fill powder & then immediately fill
TSB Broth 5.3 ml in each vial through Automatic Media Filling Nozzle fitted with
compounding vessel as per SOP-04-012. After Both filling, plug & seal the
container.

8.6.7

After sealing of vials, the vials should be inverted and shake gently to ensure that
all internal surfaces of the vials are thoroughly wetted with the broth and PEG is
dissolved in Broth solution.

8.6.8

Perform sterility testing of filtered nitrogen and compressed air as per SOP-08QC-032.

8.6.9

Check the weight of vials for 250mg and 1g vial after every 45minutes as SOP04-025.

8.6.10 Expose the settle plates at specified location during filling as per SOP-08-QC-09.
8.6.11 Perform swab test at rest (before filling) & at operational (during filling) as per
SOP-08-QC-08.
8.6.12 Perform non-viable particle count at rest (before filling) & at operational (during
filling) as per SOP-08-QC-103.
8.6.13 Perform viable particle count by using centrifugal air sampler at rest (before
filling) & at operational (during filling) as per SOP-08-QC-07.

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Surge Laboratories (Pvt.) Ltd.

ASEPTIC PROCESS VALIDATION
PROTOCOL (Dry Powder
Injectable Area)

Quality Assurance Department

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 10 of 13

8.6.14 Check the microbial status of finger of operators after filling as per SOP-08-QC026 then disinfect immediately (70% IPA) inside SS-bowl. This activity should be
performed outside the filling room.
8.6.15 Check the microbial status of clothing of operator after filling as per SOP-08-QC094. This activity should be performed outside the filling room.
8.6.16 Transfer all filled vials to QC for incubation and inspection. Incubate all vials as
in section 9.1.
8.6.17 After Broth Fill Trial, thoroughly wash/ clean, disinfect & fumigate for two
consecutive days as per SOP-04-001 & SOP-04-045
8.6.18 Now perform swab test as per SOP-08-QC-08 & expose settle plates as per SOP08-QC-09.

9.0 INCUBATION & INSPECTION:

After Filling & Sealing of Vials for 250mg & 1g, transfer all units to
Microbiology Section for Incubation & Inspection.

First invert the vials then incubate in straight position.

Incubation Conditions (First incubate at 30-35 oC for 7 days and then incubate at
20-25 oC for next 07 days)

Frequency of checking (0, 7 & 14th Day -100% while daily visual check on
random basis) in Light NLT 2000 Lux.

The Broth should be Clear in Vials-No Turbidity should be observed.

The Vials for +ve and ve test should also be checked

Record all observation on Material / Product Sterility Test Report (F-08-008), if

turbidity is observed in more than acceptable limits then Broth Fill is considered
as Failure of Aseptic Process.

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Surge Laboratories (Pvt.) Ltd.

ASEPTIC PROCESS VALIDATION
PROTOCOL (Dry Powder
Injectable Area)

Quality Assurance Department

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 11 of 13

Perform Identification test for contamination observed & thorough investigation

for the failure of run as per SOP for Handling of Out of Specification (OOS)
{SOP-08-QA-027}.

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Surge Laboratories (Pvt.) Ltd.

ASEPTIC PROCESS VALIDATION
PROTOCOL (Dry Powder
Injectable Area)

Quality Assurance Department

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 12 of 13

10.0 Deviation & Change Control:

Any Deviation (s) & Change Control from the pre approved Protocol must be recorded
investigated & justified as per SOP-08-QA-022.

11.0 Data Analysis:

Collect the data during execution of Protocol for all listed critical parameters & then
evaluate the data in the light of acceptable criteria to conclude whether the process is
validated or not, related formats for data collection as under:Sr. no.

Form Title

Form No.

Aseptic Validation Report

F-08-028

Material / Product Sterility Test Report

F-08-008

Microbial Bioburden Report

F-08-026

Sterility Testing of Nitrogen / Compressed air

F-08-021

Efficacy Testing of Disinfectants

F-08-019

Operators Breathing Test

F-08-022

Pressure Differential Monitoring

F-05-08

CFM Monitoring

F-08-001

10

Vial Washing Record

F-04-11

11

Sterilization Cycle Log

F-04-23

12

InProcess Control Record

F-04-17

13

InProcess Control Record (Vial Washing)

F-08-QA-22

14

Check List for Dry Powder Injectable Filling Area

15

InProcess Control Record (Optical checking)

F-08-QA-054
F-08-QA-23

16

InProcess Control Record (Weight )

F-08-QA-10

17

Surface Swab test report

F-08-037

18

Surface area monitoring of vial filling area

F-08-36

19

Air flow pattern report

F-08-030

20

Air particle count

F-08-002

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Surge Laboratories (Pvt.) Ltd.

ASEPTIC PROCESS VALIDATION
PROTOCOL (Dry Powder
Injectable Area)

Quality Assurance Department

Micro-10-QA-002
Revision No.: - 0
Issue Date: 25-09-2009

Page 13 of 13

21

Ampoule monitoring using ampoule indicator

F-08-027

22

Temp & Humidity Record

23

Bioburden Check for Ampoules and Vials after Washing

RG-QC-26

24

Sterility testing of empty vials/ampoules/rubber stoppers after Sterilization

RG-QC-28

25

Monitoring of Dry Heat Oven using Spore Strip Bioindicator

RG-QC-20

26

Growth Promotion Test

RG-QC-19

27

Sterility Testing of Nitrogen and Compressed Air

RG-QC-29

F-08-QA-031

12.0 RE-VALIADTION
12.1 Re-validation includes:

On Going Simulation will be performed after each six (06) months.

After Major Changes in manufacturing procedure
After Major Changes in the Area/HVAC System.

13. CONCLUSION & ISSUANCE OF CERTIFICATE

In the light of above acceptance criteria, conclude that whether Aseptic Filling Process is
Satisfactory or need some amendments . QA will issue Certificate of Successful Aseptic
Process Validation for Dry Powder Injectables.

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