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php/journal/index AJPRD 2019; 7(1): 50-61

A
J Asian Journal of Pharmaceutical Research and Development
P Open Access to Pharmaceutical and Medical Research
R
D © 2013-18, publisher and licensee AJPRD, This is an Open Access article which permits unrestricted
non-commercial use, provided the original work is properly cited

Original Article
Validation of Sterile Water for Injection in
Pharmaceutical Industry and Othersterile Facility
Choudhary Neetu*1, Patil Bhagyashri1, Choukse Raju1, Varma Ajit Kumar2, Bairagee
Deepika 2, Kulkarni Sweta 3
1
Schoolof Pharmacy, Dr.A.P.J. Abdul Kalam University, Dewas Bypass road, Indore (M.P.)
2
Orienral College of Pharmacy & Research, Oriental University, Sanwer Road, Opp. Rewati Range, Gate No-1 Jakhya, Indore,
(M.P.) India.
3
Chameli Devi Institute of Pharmacy, Khandwa Road ,Indore, Madhya Pradesh, India.

ABSTRACT

Sterile facilities for all pharmaceutical product specially to parentral preparation, is a must important back bone of
sterile formulation andor pharmaceutical dosage form. There is most important to sterile of the areas where the
formulation process proceed from initial to final stage. The sterile injectable products are very critical and sensitive
products as they are administered directly into blood circulation. These products are designed such that it should be
free from micro-organisms, pyrogens and unacceptable particulate matter. Any failure in quality and purity of these
products may directly affect the safety of patient being treated. FDA, WHO, ISO and Good Manufacturing Practiceshas
established the guides to the development of sterile pharmaceutical preparation facilities for health care
establishments.This report covers all summaries that the three batches of Methylcobalamine injection 2 ml have been
validated with the support of process validation protocol.

Keyword: USFDA, SOPS, CGMP, HVAC, ICFU,FPM.

A R T I C L E I N F O: Received: 18 Oct. 2018; Review Completed: 20 Jan.2019; Accepted: 9 Feb .2019; Available online: 15 Feb. 2019

Cite this article as:

Neetu Choudhary* ,Bhagyashri Patil ,Raju Choukse, Sweeta Kulkarni, Ajit Kumar Varma,Deepika Bairagee, Validation of
Sterile Water for Injection Inpharmaceutical Industry and Othersterile Facility, Asian Journal of Pharmaceutical Research and
Development. 2019;7(1):50-61
DOI: http://dx.doi.org/10.22270/ajprd.v7i1.446
*Address for Correspondence
Neetu Choudhary*, Schoolof Pharmacy, Dr.A.P.J. Abdul Kalam University, Dewas Bypass road, Indore (M.P.)

INTRODUCTION: and palatable dosage form of drug administration, the

prospective process validation could be easily and

V alidation is a concept that has been evolving

continuously since its first formal appearance
in the United States in 1978. Validation as it is
known today has developed from the need to maintain
thoroughly studied on this topic. Methylcobalamine is
used to produce red blood cells in pernicious anemia
and to maintain the good health.
quality, consistency and above all public safety. The Types of the validation (6,7):
present project reflects the current trends and serves as
Process validation:-
an educational tool in our progressive industry1.
It is conducted during the manufacturing process of the
Definition (USFDA): “Process validation is
product.
establishing documented evidence that provides a high
degree of assurance that a specific process will Types of process validation:-
consistently produce a product meeting its
predetermined specifications and quality characteristics  Prospective validation
(2,3)
”. Since Methylcobalamine injection (500 mcg) is a  Concurrent validation
new formulation which is going to be administered in  Retrospective validation
the form of IM route for the instant effect.The  Revalidation
injectable form is easily accepted, safe, user friendly

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Validation process (4.5) – flow diagram:-

Pre-Validation Activities

Validation Protocol Preparation

Validation Protocol-Review & Approval

Protocol Execution

Data Analysis

Validation Report & Sign Off

Change Control
Revalidation

Fig. 1.1: Validation Process- Flow Diagram

Equipment validation:- (Qualification)

The equipment should be designed and/or selected as per the product specifications are consistently achieved.

Change
New Facility, Utility/ Equipment
Control

Construction & Installation

Calibration
Program
Qualification or Requalification

QA/Safety Approval to use

Preventive
Maintenance Maintained in “Qualified” state

Close out Qualification Package

Extended
Monitoring
Facility, Utility and/or
Equipment Retired from service

Fig.1.2 Qualification Life Cycle

8,9
Types of Equipment Qualification Installation Qualification (IQ):
Design Qualification (DQ): It verifies the installations such as machines, measuring
devices, utilities, manufacturing areas used in a
It is the documented verification of the proposed design
manufacturing process.
of the facilities, systems and equipment for the intended
purpose. It involves following parameters:make, type, Operational Qualification (OQ):
model number, material of construction, size and shape
OQ checks the facilities, systems and equipment that
of different parts of the equipments.
are operating with standard conditions. It tests whether
or not the system works as expected.

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Performance Qualification (PQ): Validation of Utility:

PQ is the documented verification that the facilities,  HVAC system (AHU):
systems and equipment can perform effectively to  HEPA filter integrity test (DOP test).
perform approved process and deliver product  Air velocity across HEPA filter.
specification consistently  Air changes per hour.
Analytical method validation:  Non viable and viable particle count.
 Decontamination time.
Method validation defined as, “The process by which, it  Temperature and humidity monitoring.
is established by laboratory studies, that the
 Air flow pattern.
performance characteristics of the method meet the
Validation of Equipment:
requirements for the intended analytical application".
A) Autoclave validation.
Cleaning validation:-
B) Ampoule sterilizing tunnel validation
Cleaning validation is a process of attaining and
documenting sufficient evidence to give reasonable MATERIALS AND METHODS:
assurance given the current state of Science and
Materials:
Technology.
Drug-Methylcobalamine Injection- 2 ml
The whole plan of validation of sterile facility is
Strip- Bacillus stearothermophilus spore strips
divided into following steps10,11.
Strip- Chemical integrator strips (Steam –Clox Cards)

Table 1.1: List of Equipments

Sr. No. Equipments Manufacturer

1 Weighing balance Motter Toledo
2. Ampoules washing machine Pyroklenz
3. Autoclave Metalchem industries
4. Ampoule sticker labeling machine Maharshi Udyog
5. Ampoules filling machine Kembert
6. Ampoules sterilizing tunnel Klenzaieds
7. Filter integrity test apparatus Global Eng.
9. Particle counter Met one
10. Carton Packing machine Pam-Pac120(Hi-Cart machine)

METHODS:- filters). Velocity usually increase the possibility of

contamination as these can have an effect on
VALIDATION OF HVAC SYSTEM (Heating
unidirectional airflow in validation.
Ventilation and Air Conditioning system):
Air changes per hour:
To regulate room temperature, humidity and air flow
ensuring that such elements remain within their To evaluate the air is exchangedwith fresh or filtered air
acceptable ranges is the primary use of HVAC. in each hour (numbers of time).
DOP Test: The air changes is calculated in following ways
The purpose of performing regularly scheduled leak Non- viable and viable particulate count:
tests, also to detect leaks from the filter media, filter Environmental monitoring
frame or seal.Leak tests should be performed at suitable
Its include testing of patticle count (number of particles
time intervals for HEPA filters in the aseptic processing
per volume of air)!of various surfaces for
facility.
microbiological quality.
Air Velocity Measurement:
No. of location = √Area
To conduct periodic monitoring of uniformity of
velocity across the filter (and relative to adjacent

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Table 1.2- Air Classification

Grade Class USFDA ISO Designation 0.5μm/cu ft 5μm/cu ft.
A 100 M3.5 5 100 par/ cu ft 0 par/ cu ft.
B 1000 M4.5 6 1000par/ cu ft 7 par/ cu ft.
C 10000 M5.5 7 10000 par/ cu ft 70 par/ cu ft.
D 100000 M6.5 8 100000 par/ cu ft 700 par/ cu ft.
*Note: par/cu ft- Particles per cubic feet
VALIDATION OF EQUIPMENT:

Dispensing of raw material

Washing, sterilization &drying of accessories

Preparation of drug solution in mixing

vessel

Filtration

Washing & sterilization ofampoules in

washing machine and sterilizing tunnel

Filling and sealing

HPHV leak test

Visual inspection

Labeling

Packing

Fig 1.3: Process flow chart of manufacturing operation

Table 1.3: Manufacturing Critical Control Parameter

Test description Limit
In let WFI temperature 80±5 ºC
Cooled WFI temperature NLT 35 ºC
Nitrogen pressure NLT 5 kg/cm2
Bubble point of membrane filter NLT 2.5 kg/cm2
pH 7.2 to 7.5
Final mixing time NMT 30 min
Nitrogen purging Whole process

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Table 1.4: Machine critical control parameter

Test description Limit
Washing machine
Recycled water NLT 1.5 kg/cm2
Compressed air NLT 1.5 kg/cm2
WFI NLT 1 kg/cm2
Tunnel
Sterile zone temperature NLT 280 ºC
Pressure differential
Sterile zone 20 pascal
Cooling zone 12 pascal
Autoclave
Temperature 121±1 ºC
Steam pressure 1.2 kg/cm2
Vacuum pressure(leak test) -0.600 bar

Table 1.5: Validation of In Process Parameter

Stages Test/ Process Parameters Limit

Rawmaterial verification Balance calibration Calibrated

RM weight verification Verified
WFI pH 5-7
Conductivity <1.3µs/cm2
Bioburden 10 CFU/100ml
BET <0.25 EU/ml
Clean steam BET < 0.25 EU/ml
Total bacterial count 10 CFU/100ml
Washing Before washing bioburden <10CFU/ampoule
After washing bioburden <1 CFU/ampoule
After sterilization Bacterial endotoxin <0.25 EU/ml
Sterility after depyrogenation <1 CFU/ampoule
Set temperature of tunnel >280 ºC
Conveyor speed 72 mm/min
Mfgpreparation of drug Bioburden of drug solution < 100 CFU/ml
solution pH 7.2-7.5
Mixing efficiency 10 min (90-110%)
Temperature 40-50ºC
Filtration Bioburden < 4 CFU/100ml
Sterility No growth
Pre integrity pressure NLT 2.5kg/cm2
Post integrity pressure NMT 3.2 kg/cm2
Filter duration NMT 2 hour
Pressure for filtration 1.2kg/cm2
Compressed air and Bioburden < 1CFU
nitrogen gas Sterility No Growth
Stages Test / Process Parameters Limit
Filling and sealing Volume of ampoules 2-2.2 ml
Sealing OK
Nitrogen flushing OK

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Visual inspection OK
Sterility No Growth
Start filling Sterility No Growth
pH 7.1-7.2
Assay 90 -110%
Middle filling Sterility No Growth
pH 7.1-7.2
Assay 90-110%
End filling Sterility No Growth
pH 7.1-7.2
Assay 90-110%
HPHV leak test Leak test time NMT 15 min
Rejected ampoules LT 1%
Visual inspection Clarity OK
Output Ok
Labeling Clarity of over printing w.r.t. output OK
Packing Sealing temperature 170 ºC
Leak test OK
Clarity of over printing w.r.t. blister per minute Clear
Finished goods analysis Sterility No Growth
Assay 90-110%
Yield Filling yield NLT 90%
Packing yield NLT 90%
Visual inspection NLT 90%
Batch yield NLT 90%

Table 1.6: Worst Case Study (Bracketing Method)

Stages Assets Test Parameters Limit

Filling line speed at 150 Washing machine Particulate matter Absent

amp/min Breakage NMT 1%
No. of break down No major break down
Tunnel depyrogenation Sterilit No Growth
residence time NLT 3min Endotoxin < 0.25EU/ml
No of breakage down No major break down
Volume of filled ampoules 2 to 2.2 ml
Filling machine Sealing defect <1%
Particulate matter Absent
Break down No major break down
Washing machine Particulate matter Absent
Filling line Speed at 250 Breakage NMT 1%
ampoules/min No of break down No major break down
Tunnel depyrogenation Sterility No Growth
residence time NLT 3min Endotoxin < 0.25EU/ml
No of breakage down No major break down
Filling machine Volume of filled ampoules 2 to 2.2 ml
Sealing defect <1%
Particulate matter Absent
Break down No major break down

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RESULT& DISCUSSION:
DOP Test:
Acceptance Criteria: The leakage should not be more than 0.01%
Air Velocity Measurement:
Table 1.7: Air Velocity Result
Room No. Room Name Class Filter No. Velocity(FPM) Average
Velocity(FPM)
V1 V2 V3 V4 V5
PG1.107 Ampoule B AHU-29/PG1.107/S/01 102 95 92 89 94 94.4
Filling AHU-29/PG1.107/S/02 87 95 94 90 85 90.2
AHU-29/PG1.107/S/03 101 85 94 87 91 91.6
AHU-29/PG1.107/S/04 94 87 94 101 96 94.4
AHU-29/PG1.107/S/05 92 95 98 101 97 96.6
AHU-29/PG1.107/S/06 95 101 85 94 89 92.8
AHU-29/PG1.107/S/07 104 102 95 97 100 99.6
AHU-29/PG1.107/S/08 95 106 101 88 87 95.4
Acceptance Criteria: Average velocity must be in range of 90±20% FPM.
Calculation of Air Changes:
RoomName:Filling area
Room Volume:2160 .86 Cu ft
Area of Filters: 2 ft X 2 ft= 4 Sq ft
CFH: Average velocity X area of filter X 60 min

Table 1.8: Calculation of Air Changes

Filter No. Average Velocity(FPM) CFH
AHU-29/PG1.107/S/01 94.4 22656
AHU-29/PG1.107/S/02 90.2 21648
AHU-29/PG1.107/S/03 91.6 21984
AHU-29/PG1.107/S/04 94.4 22656
AHU-29/PG1.107/S/05 96.6 23184
AHU-29/PG1.107/S/06 92.8 222272
AHU-29/PG1.107/S/07 99.6 23904
AHU-29/PG1.107/S/08 95.4 22896
∑ CFH 181200
Air changes per hour = ∑ CFH ÷Room Volume in Cu ft
= 181200÷2160.86
=83.85 air changes
Acceptance Criteria: Min 25 air changes per hour.
Viable Particle Count: Settle Plate Method
Table 1.9: Viable Particle Count
Sr. No. Location Grade No. of Samples Count /plate

L1 L2 L3 L4 L5
1. Under LAF A 2 <1 <1 - - -
2. Filling Room B 5 <1 <1 <1 <1 <1
3. Filtration Room B 4 <1 <1 <1 <1 -
4. Cooling Zone B 3 <1 <1 <1 - -
5. Leak Test Room B 3 <1 <1 <1 - -

Pressure Differential:
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Table 1.10: Pressure Differential

Area w.r.t. area Diff Pressure Limit

Morning Reading Evening Reading

Filling Vs Filling Corridor 9.00 A.M. 8 6.30 P.M. 8 NLT 6 Pa
Cooling Vs Cooling Corridor 9.05 A.M 8 6.35 P.M. 8 NLT 6 Pa
Filling Vs Staging 9.10 A.M. 18 6.40 P.M. 18 NLT 15 Pa
Filtration Vs Sterile Corridor 9.15 A.M. 20 6.45 P.M. 20 NLT 15 Pa
Amp Filling Vs Amp washing 9.25 A.M. 18 6.55 P.M. 18 NLT 15 Pa

Temperature and Humidity Monitoring:

Room Name: Filling Room
Table 1.11: Temperature and Humidity Monitoring
Time Temperature Humidity Limit
10.00 A.M. 21.3ºc 53% Temp:23±2ºc Humidity:NMT55%
3.00 P.M. 24.5ºc 49% Temp:23±2ºc Humidity:NMT55%
6.00 P.M. 22.6ºc 47% Temp:23±2ºc Humidity:NMT 55%

Validation of the Sterilization Process in Autoclave:

Table 1.12: Temperature recorded in Autoclave
Sterilization RTD1 RTD2 RTD3 RTD4 RTD5 RTD6 RTD7 RTD8
time
(ºC) (ºC) (ºC) (ºC) (ºC) (ºC) (ºC) (ºC)

10:31:01 121.2 121.4 121.3 121.5 121.4 121.2 121.6 121.5

10:32:02 121.8 121.9 121.7 121.9 121.8 121.6 121.8 121.7
10:33:01 121.6 121.7 121.7 121.9 121.8 121.7 121.7 121.6
10:34:02 121.5 121.5 121.6 121.7 121.8 121.6 121.5 121.4
10:35:01 121.4 121.6 121.7 121.6 121.8 121.5 121.6 121.5
10:36:01 121.6 121.7 121.7 121.5 121.6 121.4 121.5 121.6
10:37:01 121.5 121.6 121.7 121.4 121.4 121.3 121.2 121.4
10:38:01 121.4 121.7 121.6 121.5 121.5 121.4 121.3 121.3
10:39:01 121.5 121.6 121.7 121.4 121.2 121.3 121.4 121.5
10:40:01 121.6 121.7 121.6 121.3 121.3 121.2 121.5 121.6
10:41:01 121.7 121.8 121.7 121.4 121.5 121.4 121.6 121.7
10:42:01 121.6 121.9 121.8 121.6 121.6 121.7 121.8 121.9
10:43:01 121.7 121.6 121.5 121.5 121.4 121.6 121.7 121.8
10:44:01 121.8 121.7 121.7 121.4 121.3 121.5 121.3 121.5
10:45:01 121.6 121.5 121.3 121.5 121.2 121.2 121.2 121.6
Average 121.6 121.7 121.6 121.5 121.5 121.4 121.5 121.6
MIN. (ºC) 121.2 121.4 121.3 121.3 121.2 121.2 121.2 121.3
MAX. (ºC) 121.8 121.9 121.8 121.9 121.9 121.7 121.8 121.9
Coolest point 121.2ºC

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Table 1.13: Manufacturing critical control parameter

Test description Batch No. X Batch No. Y Batch No. Z
In let WFI temp 84.2 ºC 84.0ºC 83.2ºC
Cooled WFI temp 28.3ºC 26.2ºC 28.00ºC
Nitrogen pressure 5.0 kg/cm2 5.2 kg/cm2 5.3 kg/cm2
2 2
Bubble point of membrane filter 3.0 kg/cm 3.2 kg/cm 3.3 kg/cm2
pH 7.9 7.8 7.9
Final mixing time 30 min 30 min 30 min
Nitrogen purging Whole process Whole process Whole process

Table 1.14: Machine critical control parameter

Test description Batch No. X Batch No. Y Batch No. Z
Washing machine
Recycled water 2.0 kg/cm2 2.0 kg/cm2 2.0 kg/cm2
Compressed water 2.0 kg/cm2 2.0 kg/cm2 2.0 kg/cm2
WFI 1.2 kg/cm2 1.2 kg/cm2 1.2 kg/cm2
Tunnel
Sterile zone temp(ºC) 330,328,326,324 330,328,326,324 330,328,326,324
Pressure differential
Sterile zone(pa) 23 26 23
Cooling zone(pa) 14 14 14
Autoclave
Temp.(ºC) 121.4 121.3 121.4
Steam pressure(kg/cm2) 1.2 kg/cm2 1.2 kg/cm2 1.2 kg/cm2
Vacuum pressure(Leak test) -0.600 bar -0.600 bar -0.600 bar

Table 1.15: Validation of In Process Parameter Result

Stage Test/Process parameter Result

Batch No. X Batch No. Y Batch No. Z

Raw material weight Balance calibration Calibrated Calibrated Calibrated
verification RM weight verification Verified Verified Verified
WFI Bacterial endotoxin <0.25 EU/ml <0.25 EU/ml <0.25 EU/ml
pH 6.17 5.44 5.67
Bioburden <1CFU/100ml <1CFU/100ml <1CFU/100ml
2 2
Conductivity 0.5423µs/cm 0.7483µs/cm 0.7463µs/cm2
Clean steam Bacterial endotoxin <0.25 EU/ml <0.25 EU/ml <0.25 EU/ml
Total bacterial count <1CFU/100ml <1CFU/100ml <1CFU/100ml
Ampoule washing & Before washing bioburden 03 CFU/ampoule 07 CFU/ 05 CFU/ ampoule
sterilization/depyroge Particulate matter absent ampoule
absent absent
nation
After washing bioburden <1CFU/amp <1CFU/amp <1CFU/amp
After sterilization/depyrogenation
Bacterial endotoxin <0.25 EU/ml <0.25 EU/ml <0.25 EU/ml
Sterility No growth No growth No growth
Set temp. of tunnel(ºC) 330,328,326,324 330,328,326,324 330,328,326,324
Conveyour speed 67 mm/min 67 mm/min 67 mm/min
Mfgpreparation of Bioburden of drug solution 2 CFU/100ml 4 CFU/100ml 3 CFU/100ml
drug solution pH 7.2 7.1 7.1
Mixing efficiency 100.1% 96.5% 98.5%

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Filtration Temperature 32 ºC 33 ºC 35 ºC
Sterility No growth No growth No growth
Post integritypressure
Pre integrity pressure 3.2 kg/cm22
2.5 kg/cm 3.2 kg/cm22
2.5 kg/cm 3.2
2.5 kg/cm
2
kg/cm2
Filter duration 60 min 75 min 70 min
Bioburden <1CFU <1 CFU <1 CFU
2 2
Pressure for filtration 3.2 kg/cm 3.2 kg/cm 3.2 kg/cm2
Compressed air Bioburden <1 CFU <1 CFU <1 CFU
&nitrogen gas Sterility No growth No growth No growth
Filling and sealing Volume of ampoule 2.2 ml 2.2 ml 2.2 ml
Sealing OK OK OK
Nitrogen flushing OK OK OK
Visual inspection(rejection) 07 04 08
Sterility No growth No growth No growth
HPHV leak test Leak test time 11 min 12 min 10 min
Rejected ampoule 05 08 07

Assay of Methyl Cobalamine during start, middle and end of filling:

Table 1.16: Assay of Methyl Cobalmine
Series Assay of Methyl Cobalamine (%)
Batch No.
X Y Z
S M E S M E S M E
01 100.72 100.73 104.45 100.64 98.62 97.32 99.71 98.72 101.78
02 100.65 101.28 100.06 99.47 100.82 99.76 100.33 98.21 100.62
03 99.22 100.25 101.44 100.05 101.17 98.07 100.94 97.44 100.16
04 101.34 102.14 100.05 98.15 98.82 101.36 100.86 97.94 100.19
05 101.09 100.39 101.49 100.74 101.12 99.91 99.54 100.14 99.03
06 100.56 100.79 99.09 98.05 102.07 99.27 99.06 97.67 100.96
07 101.08 100.47 98.94 102.35 101.22 100.41 100.11 99.11 100.66
08 99.91 100.96 98.82 100.79 98.47 99.12 100.74 98.35 100.95
09 100.22 101.37 101.83 98.29 98.67 102.01 100.28 99.61 100.13
10 101.04 99.73 99.78 98.68 101.32 98.27 101.34 97.98 99.44
Max 101.34 102.14 104.45 102.35 102.07 102.01 100.94 100.14 101.78
Min 99.22 99.73 98.82 98.05 98.47 97.32 99.06 97.44 99.03
Mean (%) 100.58 100.87 100.60 99.72 100.23 99.55 100.29 98.52 100.39
% RSD 0.84 0.67 1.72 1.44 1.40 1.47 0.71 0.87 0.79

Fig 1.4: Assay of Methylcobalamine series 1-3 Fig 1.5: Assay of Methylcobalamineseries 4-6

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Fig1.6: Assay of Methylcobalamineseries 7-10

Filling Line Speed Validation at 150 ampoule/min

Table 1.17: Result of filling line speed validation at 150 ampoule/min
Assets Test parameter Batch No
X Y Z
Washing Particulate matter No particulate matter No particulate matter No particulate
machine Breakage LT 1% LT 1% matter
LT 1%
No. of break down No major break down No major break down No major break
Tunnel Sterility No growth No growth down
No growth
Endotoxin <0.25EU/ml <0.25EU/ml <0.25EU/ml
Breakage LT 1% LT 1% LT 1%
No. of breakage down No major break down No major break down No major break
Filling Volume 2.2 ml 2.2 ml down
2.2 ml
machine Sealing defect LT 1% LT 1% LT 1%
Particulate matter LT 2% LT 2% LT 2%
Break down No major break down No major break down No major break
Labelling Coding on label OK OK down
OK
machine Breakage LT 1% LT 1% LT 1%
No .of break down No major break down No major break down No major break
Cartooning Coding on carton OK OK down
OK
machine Breakage LT 1% LT 1% LT 1%
No .of break down No major break down No major break down No major break
down

Filling Line Speed Validation at 250 ampoule/min

Table 1.18: Result of filling line speed validation at 250 ampoule/min
Assets Test parameter Batch No.

X Y Z
Washing Particulate matter No particulate matter No particulate matter No particulate matter
machine Breakage LT 1% LT 1% LT 1%
No. of break down No major break down No major break down No major break down
Tunnel Sterility No growth No growth No growth
Endotoxin <0.25EU/ml <0.25EU/ml <0.25EU/ml
Breakage LT 1% LT 1% LT 1%
No. of breakage down No major break down No major break down No major break down
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Choudhary et al Available online at http://ajprd.com/index.php/journal/index AJPRD 2019; 7(1): 50-61

Filling Volume 2.2 ml 2.2 ml 2.2 ml

machine Sealing defect LT 1% LT 1% LT 1%
Particulate matter LT 2% LT 2% LT 2%
Break down No major break down No major break down No major break down
Labelling Coding on label OK OK OK
machine Breakage LT 1% LT 1% LT 1%
No .of break down No major break down No major break down No major break down
Cartooning Coding on carton OK OK OK
machine Breakage LT 1% LT 1% LT 1%
No .of break down No major break down No major break down No major break down
SUMMARY:
Validation of HVAC system ensures that all these parameter are within the predetermined specification.
Test/Critical parameter Acceptance criteria
DOP test NMT 0.01%
Air velocity 90±20 % FPM
Air changes NLT 25 air changes
Pressure differential For same class NLT 6 Pa and different class NLT 15 Pa
Temp and humidity Temp:23±2ºc , Humidity:NMT55%
Non-viable count As per ISO specification
Viable count As per IHS guideline
Air flow pattern Uniform up to the operational level
Decontamination time NMT 8 minutes

CONCLUSION: criteria. For the intended indication of new drug (

accurate and reliable assessment) for its effectiveness
Based on the validation test results, review, assessment
and safety, it is necessary before approval of new drug
and evaluation it is concluded that the manufacturing
Pharmaceutical validation and process control are
process of Methylcobalamine injection is validated (as
required facilities.
per cGMP guidelines) for the predetermined acceptance
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ISSN: 2320-4850 [61] CODEN (USA): AJPRHS