PROCESS VALIDATION

PROTOCOL

FOR

CEFTRIAXONE INJECTION IP 2 g

1. List of contents:

Sr. Contents
No.
Protocol Contents
1
Protocol Approval
2
Objective
3
Scope
4
Responsibility
5
Process
6
Acceptance Criteria
7
Methods for Recording and Evaluating Results
8
Observed Deviation
9
Qualification Protocol
20
Approval of Qualification Protocol
22
Re-Validation Criteria
22
Validation Report
23
Evaluation and Approval of Validation Report
24
Use of the Batches
25
List of Annexure /Exhibits
26
Reference Documents
27
1.0 APPROVAL OF PROCESS VALIDATION PROTOCOL:
The process validation protocol has been prepared, checked and approved for
implementation by the signed:

Department Name Signature Date

PREPARED
BY Quality
Assurance

Department Name Signature Date

Production
CHECKED
BY Quality control

Quality
Assurance

Department Name Signature Date

APPROVED
BY Quality
Assurance

2.0 OBJECTIVE:

As an important regulatory requirement, process of manufacturing of Ceftriaxone

Injection IP 2 g has to be qualified in process validation. Process validation
becomes an important integral part of the validation exercise to establish
consistency in the process of manufacturing.
 To establish documented evidence, this will provide a high degree of assurance and
reliability to produce the product with reproducible results. This procedure is
tailored to challenge the exact process or product to be validated using normal
operating procedures. Validation will demonstrate that by using these parameters,
the quality product intended can be produced consistently.

The purpose of this concurrent process validation protocol is to provide information

regarding the process parameters and the tests / checks that needed to be performed
during processing. It also provides acceptable limits / ranges that will establish
quality and constant process of manufacture.

.
3.0 SCOPE:

This protocol is applicable to the manufacturing process for the product mentioned as
under:

Name of Product: Ceftriaxone Injection IP 2 g

Claim: Each vial contains:

Sterile Ceftriaxone Sodium IP
Eq. to Anhydrous Ceftriaxone 2000 mg

4.0 RESPONSIBILITY:
DEPARTMENT ACTIVITY RESPONSIBILITY

Preparation of process validation protocol Officer

Approval of process validation protocol Head

QUALITY
ASSURANCE Planning & organizing and implementing process
Head
validation of three consecutive batches
To withdraw the samples as per sampling program Officer
 Evaluation of analytical reports Head

To review the validation protocol Head

Analysis of collected samples and preparation of
Officer
QUALITY the analysis report
CONTROL To check the results and submission of analytical
Officer
report to QA
To review the validation report Head

To review the validation protocol Head

Conducting Process validation of 3 consecutive
Officer
batches
PRODUCTION To ensure performance of activities as per protocol Officer / Sr. Officer

To co-ordinate the activity Head

To review the validation report Head

5.0 Process:
5.1 Process Validation Methodology
5.1.1 During the course of validation the documentation systems, laboratory
controls, in process checks and filled vials shall be evaluated.
5.1.2 The Process Validation Methodology shall consist of three basic parts: process
parameters monitoring, the routine in process and final product release testing &
additional validation sampling and testing.
5.1.3 The type of validation to be carried out is concurrent validation as the process is
well understood and the area and equipment to be used have been validated
previously.
5.1.4 The validation exercise for three consecutive batches of production scale will be
done to assess the process consistency and the individual batches should have
raw material (API) of same manufacturing batch number.
 5.1.5 The validation protocol requires pre-approval for sampling plan and acceptance
criteria before execution.
5.1.6 At the conclusion of the process validation, a capability study will be performed
to determine whether the process is consistent and capable.
5.1.7 In order to measure these two process characteristics will be established and
evaluated to provide a high degree of assurance that the manufacturing process is
capable of repeatedly and reliably producing a finished product of the required
quality.
5.1.8 Product Summary
5.1.9 Certificate of analysis of all following materials shall be included in the
validation protocol.
5.1.10 A.R. No. and quantity per batch to be verified and recorded in the validation
protocol.
5.2 Batch Details
5.2.1 Product Name: Ceftriaxone Injection IP 2 g
Composition:
Each vial contains:
Sterile Ceftriaxone Sodium IP
Eq. to Anhydrous Ceftriaxone 2000 mg

5.2.2 Three batches are studied of equal Batch Size and Shelf Life and details of
batches as per Annexure – I.

5.3 Validation Procedure:

5.3.1 Process flow chart.

DECARTONING OF
VIALS

2. Load
2. Soaking Time INSPECTION OF
3. Rinsing with VIALS
Speed
Purified Water
Rubber Bung Particle Count
4. Rinsing with
washing Sterility
WFI WASHING OF VIALS Endotoxin
5. Siliconization
Time
Sterilization 2. Conveyer
of Rubber DE-PYROGENATION Speed
2. Load Bung OF VIALS 2. Sterilization.
2. Sterilization Temp.
Time
3. Sterilization Sterilization
Temp. of Aluminum FILLING & SEALING Speed of
4. Pressure Seal filling &
5. Vacuum Sealing M/C
Drying
 Sterilization
of Machine OPTICAL INSPECTION

LABELING & PACKING

FINISH GOODS STORE

5.4 Description of Equipment Involved in Manufacturing Process

5.4.1 All the major equipments used for process, as listed below have been verified for
Installation, Operational and Performance Qualification.
5.4.2 Integrity testing of HEPA filter has been verified.
5.4.3 Standard operating Procedure for Filter Cleaning, Maintenance, Integrity testing
of HEPA filters and sanitization of water system has been verified.

Equipment
System Description ID No.

Vial Washing M/C

Depyrogenation Tunnel
Bung processor cum Autoclave
Dry injection Filling, Rubber Stopper

Vial Sealing Machine

5.5 MANUFACTURING, ENVIRONMENT AND PRODUCT

SENSITIVE FACTORS:
Batch no.->
Area Temp.(0C) %RH Temp.(0C) %RH Temp.(0C) %RH
Raw material
dispensing
Vial washing &
depyrogenation
Vial filling &
stoppering
Vial sealing
packing

*Refer attached annexure-II

5.6 RAW & PRIMARY PACKING MATERIAL DETAILS:
5.6.1 RAW MATERIAL DETAILS:

Sr. Name of the raw Specification A.R. no. Manufacturer/

No. material supplier
2. Sterile Ceftriaxone
Sodium IP

5.5.2 PRIMARY PACKING MATERIAL DETAILS:

Sr. No. Name of the packing A.R. no. Manufacturer/supplier
material
2. 20ml moulded glass vials
USP type III
2. 20mm grey butyl rubber
stopper
3. 20mm Blue color Flip Off
Seals
* Details of Raw Material and Packing Material as per Annexure – III.

5.7 Reference Documents

5.7.1 Master Documents:
5.7.1.1 Master document shall be approved, signed and dated by appropriate and
authorized persons prior to commencement of the batch processing.
5.7.1.2 Verify the following master documents.
5.7.1.2.1.1 Batch Manufacturing Record.
5.7.1.2.1.2 Raw Material Specification.
5.7.1.2.1.3 Packaging Material Specifications.
5.7.1.2.1.4 Finished Product Specification to be included in the
Validation Protocol.

5.8 Manufacturing Process Description

5.8.1 Decartoning of vials.
5.8.2 Inspection of Vials.
 5.8.3 Washing and De-pyrogenation of vials.
5.8.4 Washing, Siliconization, Sterilization and drying of rubber plugs.
5.8.5 Autoclaving of seals.
5.8.6 Washing and Sterilization of Machine Parts.
5.8.7 Weighing & verification of quantity of raw material & transfer to filling area
5.8.8 Filling and Sealing.
5.8.9 Visual Inspection.
5.8.10 In process Quality Control Checks.
5.8.11 Labeling & Packaging.
5.9 Manufacturing Details:
5.9.1 Decartoning of Vials
5.9.1.1 Dedust the cartons.
5.9.1.2 Decartoning and remove the vials.
5.9.1.3 Arrange in the SS trays and feed to the vial washing machine.
5.9.2 Inspection of Vials
5.9.2.1 Inspect the vials visually for absence of black particles, broken glasses,
fibers and other damages.
5.9.2.2 Segregate the rejected vials, and destroy.
5.9.2.3 Account for the rejection.
5.9.3 Washing and Depyrogenation of Vials.
5.9.3.1 Feed the good vials into the conveyor belt of Linear Vial Washing
Machine.
5.9.3.2 Vials are washed from inside and outside with 5 filtered recycled
Water in the first cycle followed by air.
5.9.3.3 Vials are further washed from inside and outside in the second cycle
with 5 filtered Purified water followed by air.
5.9.3.4 Finally vials are washed from inside with 0.2 filtered WFI (above
80°C) followed by filtered air.
5.9.3.5 Washed vials are passed to the conveyor of the depyrogenating tunnel.
5.9.3.6 Vials are depyrogenated at 320°C – 360°.
5.9.3.7 Depyrogenated vials are cooled, stabilized and passed directly to the
sterile filling area.
5.9.3.8 The data shall be recorded in Annexure –IV.
5.9.4 Washing, Siliconization, Sterilization and Drying of Rubber Plugs:
5.9.4.1 Open the poly bags and remove the rubber bungs and verify physically
to ensure cleanliness.
5.9.4.2 Load the rubber bungs in S.S. perforated container and load in Bung
Processor.
5.9.4.3 Fill purified water and add Teepol for washing of bungs for 25 min.
 5.9.4.4 Then rinse and drain the purified water.
5.9.4.5 Wash the bungs with fresh purified water for 25 minutes.
5.9.4.6 Wash the bungs with Water for Injection for 20 minutes.
5.9.4.7 Do Siliconization with water for Injection for 20 minutes (200ml silicon
per load).
5.9.4.8 Sterilize thee bungs with temperature of 222ºC for 30 minutes.
5.9.4.9 Dry the bungs with Vacuum for 30 minutes
5.9.4.10Check and the bungs.
5.9.4.11 Unload the plugs in S.S. container and keep under LAF in cool zone and
take the plugs in to the sterile filling area while process is going to be
start.
5.9.4.12The data shall be recorded in Annexure – V.
5.9.5 Sterilization of Flip-Off Aluminium Seals.
5.9.5.1 Open the poly bags and remove the Aluminum seal and verify physically
to ensure cleanliness.
5.9.5.2 Load the Aluminum seal in S.S. perforated container and load in
Autoclave.
5.9.5.3 Sterilize the Aluminum seal with temperature of 222ºC for 30 minutes.
5.9.5.4 Unload the seals in S.S. container and keep under LAF in cool zone and
take the seals in to the sterile filling area while process is going to be
start.
5.9.5.5 The data shall be recorded in Annexure – VI.

5.9.6 Sterilization of Machine Parts

5.9.6.1 Sterilize the Machine parts in Autoclave at 222C for 30 minutes. Dry
the parts under vacuum for 30 minutes.
5.9.7 Weighing & Verification of Quantity of Raw Material
5.9.7.1 Checking of Raw material: Check and note the Gross Wt., AR No, Mfg.
Date and Exp. Date of RM Container.

5.9.8 Filling and Sealing

5.9.8.1 Take the sterile powder for filling in 20 ml previously sterilized vials
under nitrogen using vial filling and capping line adjusted to calculated
fill weight.
5.9.8.2 Load the powder to the hopper of the filling machine after ensuring that
the hopper, fiber disc, porthole and Stoppering unit assembly is
sterilized.
5.9.8.3 Weight adjustment: Adjust the weight of the powder delivered into the
 sterile vials to the calculated quantity (calculated fill weight depending
on the potency). Note the time.
5.9.8.4 Vials must be closed by sterile plugs and aluminium flip-off seals.
Ensure that the plugs are tightly fitted and seal is crimped properly.
5.9.8.5 Get the vials sealed immediately.
5.9.8.6 Check vials from each porthole during filling and sealing operation for
fill weight on electronic balance at predetermined intervals.
5.9.8.7 Finally inform QA to take the sample for analysis.
5.9.9 Inspection of Vials:
5.9.9.1 Perform Optical inspection of all vials for visual inspection, breakage
and quality of sealing under Optical Inspection table.

5.9.10 In Process Quality Control Checks:

5.9.10.1 Q.C Department shall check the various parameters such as fill weight,
constituted solution, sealing integrity, etc. as per in process specification.
And record the data in Annexure –VII.
5.9.11 Labeling & Packaging:
5.9.11.1Take line clearance from Quality Assurance Officer.
5.9.11.2Start labeling and packing operation according to SOP.
5.10 Critical quality attributes summary and critical process parameters:

S.No. Process stage CPP CQA CPC Rationale for Sampling/

selection Process
verification
2. Environmental Microbial Viable Particle. Automatic Environmental Check
factors monitoring, count & Non and manual factors affect Temperature,
particle count, Viable Particle control. the product %RH,
temperature, quality. Differential
%RH & Pressure &
differential Particle Count
pressure as per limit
monitoring
throughout
the process.
2. Dispensing Temperature, Degradation, Hygrometer Product AP,
% Relative Physical nature monitoring, quality impact temperature,
humidity, degradation Pre filter & %RH, PPM
differential HEPA filter status & light
pressure (Only for light
monitoring sensitive
light material) shall
sensitivity be verified as
per BMR.
3. Vial Washing Water Clarity HMI and Visual quality Check the
Pressure manual check. Particles if any
 Compressed control. present
Air pressure
4. Depyrogenation Differential Depyrogenation BET testing BET free Vials should be
of vials pressure, sterilized
Temperature,
Conveyer
speed
5. Rubber stopper Clarity of MLT,Sterility, HMI and Product Rubber Stoppers
washing & water and BET&Moisture manual quality impact for sterilized
sterilization Sterilization content control.
temperature.
6. Filling Machine Weight HMI and weight Check the fill
speed, variation manual variation and weight
vacuum control. sterility
pressure,
dosing
pressure,
nitrogen
pressure.
7. Sealing Machine Seal integrity& HMI and Sterility and Check integrity
speed,seal clarity manual sealing of sealing &
integrity & control integrity reconstitution.
reconstitution.

5.11 Sampling Plan:

Stage Samping & Location Test Acceptance Analysis/ Sample size
criteria checked done
by
Washing Area Settle Plate <50 cfu/4hrs Micro NAP
count(cfu/4hr
s)
Settle Plate <5 cfu/4hrs NAP
Count(cfu/4h
rs)
Surface <5cfu/plate cfu/25cm2
Monitoring
at end of Micro
Operation(cf
Environmen Filling Area u/4hrs)
tal Factor Personal <5cfu/plate cfu/25cm2
Monitoring-
Gloves&
Garments(cf
u/4hrs)
Under LAF Settle Plate <2 cfu/4hrs Micro °°°
Count(cfu/4h
rs)
Before Sterilization MLT NMT 25cfu 20 No’s
Rubber
Stoppers After Sterlization Pooled Sterility,BET Should Be Micro 20 No’s
sample of initial, Middle &Moisture Sterile
 & End content
Minimum speed 60
vials/min
Washed Optimum speed 80 Clearity and Visibly Clear PR and Micro 20 vials
Vial(Before vials/min bio burden and NMT from each
Depyrogena 20cfu speed
tion) Maximum speed 200
vials/min
After Pooled sample of initial, BET NMT 0.20 Micro 3vials from
Depyrogena middle & end Endotoxin each
tion unit/mg stage(initial,
middle, end
& Pooled)
Minimum speed 50
vials/min
Filling & Optimum speed 75 22 vials
Stoppering vials/min Fill weight NMT the limit IPQA from each
Machine in BMR stage will be
speed at Maximum speed 200 checked by
three speeds vials/min IPQA
Description,
Average
content of
vials, pH,
Filling & Minimum speed 50 water Should Chemical 25 vials
Stoppering vials/min content, comply as per from each
Machine Optimum speed 75 Clarity& the speed
speed at vials/min colority of specification
three speeds the
reconstitutio
n solution &
Maximum speed 200 Particulate
vials/min matter by
visible
particle
Filling & Initial, middle & end Sterilty Should be Micro 20 vials
Stoppering sterile from each
stage(initial,
middle, end
& Pooled)
Sealing Minimum speed Sealing Should be IPQA 6 vials from
Machine 40vials/min integrity sealed each stage
Speed at will be
three speeds checked by
IPQA

6.0 ACCEPTANCE CRITERIA:

Finished product specification : AH/FPS/028/R02

S.No. Test Specification

 1. Description A white or almost white powder filled in a 20 ml
colourless glass vial, sealed with grey butyl rubber
stopper and blue coloured FLIP-OFF aluminium seal.
2. Identification Should be positive for Ceftriaxone sodium.

3. Appearance of Solution Complies

4. pH 6.0 to 8.0 (Determine in 20% w/v Solution)

5. Related substances Any individual impurity NMT 2.0 %

Total impurity NMT 5.0%
6. Clarity and colour of solution The solution should be clear and NMT intensely
coloured than reference solution B5 or BY5
7. Average fill weight ±5.0% of average weight

8. Uniformity of Average fill Not more than 2 of the individual weights deviate from
weight the average weight by more than 5 percent and none
deviation by more than 20 percent .
9. Water NMT: 22.0% w/w

10. Sterility Complies with the test for sterility

11. Particular matter
Visual particulate matter Reconstituted solution free from visual particles.
Sub-visible particle count i). Particles≥20µm Not more than 6000 particles
(By LOPC) per vials
ii). Particles≥25µm Not more than 600 particles
per vials
12. Bacterial Endotoxin test NMT 0.2 Endotoxin unit/mg

13. Assay :
Each vial contains:
Sterile Ceftriaxone Sodium Limit :90.0% to 225.0% .
IP Eq. to Anhydrous
Ceftriaxone 2000 mg

7.0 Methods for Recording and Evaluating Results:

7.1 The Protocol should be pre-approved prior to its execution and the Protocol should have
detailed sampling plan and acceptance criteria for process parameters.
7.2 QC/QA department shall take samples as per approved sampling plan.
7.3 All parameters shall be recorded in relevant records (e.g. Forms, BMR)
7.4 For Environmental parameters like viable and non-viable particles Count shall be done
as per the environmental monitoring plans.
7.5 For parameters recorded at beginning, middle and end, all values will be recorded in the
process validation report.
7.6 QC will analyze all validation samples and the raw data will be recorded / attached with
the Protocol.
7.7 Where applicable the graph and data print outs of critical process parameters will be
 obtained and attached.
7.8 Process Validation data will be recorded in the report and the recorded data will be
identified by date and signature of responsible person.
7.9 All documents related to process validation including batch specific information like and
certificates for test instruments used for analysis and certificates for critical instruments
of the processing equipment should be verified.
7.10 Raw data will be analyzed by statistical methods and the analyzed data will be presented
in the report and evaluation will be done.
8.0 OBSERVED DEVIATION:
8.1 Any deviation observed during Process Validation shall be recorded in the observed
deviation, corrective action and justification Protocol section.
8.2 Observed deviation shall be reported to the department head and quality head.
8.3 If the observed deviation does not have any major impact on the Qualification the final
conclusion shall be provided.
8.4 If the observed deviation has major impact on the Qualification, deviation shall be
Protocol to the manufacturer for the corrective action and Qualification activity shall be
redone.
8.5 The observations shall be recorded as per Annexure – VIII.

9.0 QUALIFICATION PROTOCOL:

9.1 The Process Validation Protocol shall consist of a summary document, in narrative form,
which shall briefly describe the activity performed along with the observations recorded
in relevant exhibits.
9.2 This Protocol shall also include the related documents and attachments / annexure which
were completed at the time of Qualification activity.

10.0 APPROVAL OF QUALIFICATION PROTOCOL:

The Protocol shall be evaluated and proper references / conclusions / recommendations
shall be recorded by Quality Assurance.

11.0 RE-VALIDATION CRITERIA:

11.1 If there are any significant changes to the accepted process, a re-validation of the process
must be considered.
11.2 Similarly, if there are any equipment changes, they should be evaluated against this
process validation and a determination must be made if re-validation is necessary.

12.0 VALIDATION REPORT:

 The validation report shall consists of a summary document, in narrative form, which
shall briefly describe the activity performed along with the observations recorded in
relevant exhibits which were completed at the time of qualification activity.
13.0 EVALUATION AND APPROVAL OF VALIDATION REPORT:
13.1 The report shall be evaluated and proper references / conclusions / recommendations
shall be recorded by validation team.
13.2 The validation report shall be finally approved by Head-Validations/Quality
Assurance as per Annexure – IX.

14.0 USE OF THE BATCHES

14.1 The batches will be used to perform process validation.
14.2 Results of chemical, microbial and physical tests in compliance with product
specifications will demonstrate that the process is under control, capable to deliver a
product within the expected specifications and suitable for routine production

15.0 LIST OF ANNEXURE:

Sr. No. Annexure No. of Pages

02 Batch Details for Process Validation 02
02 Manufacturing, Environment and Product Sensitive factors 02
03 Vendor Details of Raw Material and Packing Material 02
04 Vial Washing & Depyrogenation Record 02
05 Rubber Bung Process Record 02
06 Aluminum Seal Record 02
07 Process Parameters During Powder Filling & Sealing 02
08 Deviation Observed 02
09 Finished Product Analysis Report 02
20 Environment monitoring 02

16.0 REFERENCE DOCUMENTS: (If any)

16.1 Pharmaceutical Process Validation (Third Edition)
- Robert A. Nash
16.2 WHO Guideline