PROCESS VALIDATION PROTOCOL

Product Name AMIKACIN SULPHATE INJECTION , 250 mg / mL

Protocol No. DL-PV001 Page No. 1 of 6
Effective date Vial Fill Volume

ANNEXURE-5: TEST PROGRAM AND ACCEPTANCE CRITERIA

Samples shall be collected at different intervals as specified and the samples shall be analyzed in accordance with their respective test procedures
to verify that the process is under control and consistently produces a quality product. The following describes the sampling plan.

2 nd Batch
Stage Sample ID Tests Acceptance Criteria 1 st Batch Number Number
3 rd Batch Number

A clear, colorless and

Description odorless liquid
pH Between 5.0 – 7.0
WFI A Conductivity NMT 1.3µs/cm
BET Not more than 0.25 EU/mL
A.R. Number

2 nd Batch
Stage Sample ID Tests Acceptance Criteria 1 st Batch Number Number
3 rd Batch Number

A clear, colorless and

Description odorless liquid
pH Between 5.0 – 7.0
WFI A Conductivity NMT 1.3µs/cm
BET Not more than 0.25 EU/mL
A.R. Number
 PROCESS VALIDATION PROTOCOL

Product Name AMIKACIN SULPHATE INJECTION , 250 mg / mL

Protocol No. DL-PV001 Page No. 2 of 6
Effective date Vial Fill Volume

Type of the Acceptance

Stage Tests Location Sample Size Frequency Sample ID
container Criteria
Bulk solution Mixing at
 Description
from mixing About Cleaned glass 1 batch 1000
st
As per In Process
Compounding  pH B1
vessel (from 140 mL bottle RPM for 30 Specification
 Assay bottom) min
2nd batch 1500
RPM for 30
Bulk solution Cleaned
About 100 min
Compounding  Bioburden from mixing sterilized glass B2 Report the results
mL 3rd batch 2000
vessel bottle
RPM for 30
min
Pre-Integrity:
 Pre – Bubble point NLT
Integrity For capsule Before and 3180 mbar
Filter integrity NA NA NA
 Post - filters after filtration Post-Integrity:
Integrity Bubble point NLT
3120 mbar
During offline Two flushes C1
About 150 Cleaned
filtration and  Description from end of During off line
mL for each sterilized glass Report the results
filtration flush  Assay the filtration filtration
flush bottle C2
( for 2 flushes) device
 PROCESS VALIDATION PROTOCOL

Product Name AMIKACIN SULPHATE INJECTION , 250 mg / mL

Protocol No. DL-PV001 Page No. 3 of 6
Effective date Vial Fill Volume

Type of the Acceptance

Stage Tests Location Sample Size Frequency Sample ID
container Criteria
One sample
Cleaned
After offline from About 100 After off line Should be absence
 Bio burden sterilized glass D1
filtration receiving mL filtration of growth
bottle
vessel

 Description Samples from D2*

About 50 mL
 pH receiving at each
 Assay vessel at interval Initial, 24
After offline Cleaned glass
different time hours, 48 hours Report the results
filtration bottles
intervals as and 72 hours
per HTS D3*
About 100
 Bio burden protocol mL at each
interval

* Keep approximately about 500 mL of offline filtered solution in receiving vessel for real hold time study for any one of the validation batch.
collect approximately 50 mL for each time interval from receiving vessel for (chemical) and 100 mL for (Bioburden) hold time studies
Real time hold study shall be performed up to 72 hours within sample intervals of initial, 24 hours, 48 hours and 72 hours.
 PROCESS VALIDATION PROTOCOL

Product Name AMIKACIN SULPHATE INJECTION , 250 mg / mL

Protocol No. DL-PV001 Page No. 4 of 6
Effective date Vial Fill Volume

Type of the Acceptance

Stage Tests Location Sample Size Frequency Sample ID
container Criteria
Two flushes
During online E1
(each 150 mL During online
filtration and  Description During filling equivalent Filled Vials filtration and Report the results
filling flush  Assay vials shall be filling into vials E2
( for 2 flushes)
collected)
Pre-Integrity:
 Pre – Bubble point NLT
Integrity Before and 3180 mbar
Filter Integrity For filters NA NA NA
 Post - after filtration Post-Integrity:
Integrity Bubble point NLT
3120 mbar
 PROCESS VALIDATION PROTOCOL

Product Name AMIKACIN SULPHATE INJECTION , 250 mg / mL

Protocol No. DL-PV001 Page No. 5 of 6
Effective date Vial Fill Volume

Type of the Acceptance

Stage Tests Location Sample Size Frequency Sample ID
container Criteria
Sample shall
collect at F1
Initial, Middle
and End of:
1st batch -
 Description Minimum F2
Filling and  Content 10 vials speed
10 vials Filled Vials Report the results
Stoppering uniformity during filling 2nd batch -
 Assay optimum /
medium / target
speed F3
3rd batch -
maximum
speed
 PROCESS VALIDATION PROTOCOL

Product Name AMIKACIN SULPHATE INJECTION , 250 mg / mL

Protocol No. DL-PV001 Page No. 6 of 6
Effective date Vial Fill Volume

Type of the Acceptance

Stage Tests Location Sample Size Frequency Sample ID
container Criteria
Sample shall
collect at G1
Initial, Middle
and End of:
1st batch -
G2
08 vials at Minimum
 Seal integrity initial ,middle speed Should pass seal
Sealing 24 vials Sealed vials
/ Leak test and end of the 2nd batch - integrity test
sealing optimum /
medium / target
speed G3
3 batch -
rd

maximum
speed
Pooled
Optically As per FP
Optical As per FP sample after After Optical
46 Vials inspected good H Specification
inspection Specification optical inspection
vials (Release)
inspection

Note: Personnel shall wear gloves, goggles, etc. during sampling activity.