PRODUCT DOSSIER FOR THE

REGISTRATION OF PHARMACEUTICAL
PRODUCT IN NEPAL

POLY-B
(POLYMYXIN B SULPHATE INJECTION 500000 IU)

Submitted by :
SAMARTH LIFE SCIENCES PVT. LTD.
Nangal Uperla, Swarghat Road, Nalagarh,
Dist.: Solan (H.P.)

INDEX
Section
No.

Description

Manufacturing License

COPP (Certificate of Pharmaceutical Product)

Batch Formula

Product Specification

Methods of Analysis

Samples of Labels & Carton

Samples of the Product (2 unit pack)

Analytical Report

Stability Data

1. MANUFACTURING LICENSE

2. COPP (CERTIFICATE OF ANALYSIS)

COPP (CERTIFICATE OF ANALYSIS) : ATTACHED.

3. BATCH FORMULA

POLY-B (Polymyxin B Sulphate Injection 500000 IU)

Composition : Each vial contains :
Polymyxin B Sulphate BP .......................... 500000
Water for Injections IP ................................ q.s.
Description of dosage form : White lyophilized cake filled in 5 ml amber colour glass vial, labeled,
stoppered and sealed with blue colour flip off seal, such one vial packed
in PVC tray with cover, such one tray along with packing insert is packed
in a carton.
Type of container and closure :

Batch Size : 7000 Vials

Sr

Name of

No
1.

Specific

Strength

Qty

Overages

Actual Qty

Function

Ingredient

ation

per vial

Required

added

Used

Polymyxin B

BP

500000

Antibiotic

IP

q. s.

Vehicle

Sulphate
2.

WFI

3.
* q.s - Quantity Sufficient

4. PRODUCT SPECIFICATION

Specification of Finish Product :

Product

: POLY-B (Polymyxin B Sulphate Injection 500000 IU)

TEST

Discription

Identification

STANDARD LIMIT
White lyophilizedcake filled in 5 ml amber colour
glass vial, labelled, stoppered and sealed with blue
colour flip off seal, such one vial packed in PVC tray
with cover, such one tray along with packing insert is
packed in a carton.
As per specification

pH

5.0 to 7.0

Average filled weight

As per specification

Water

NMT 5.0% w/w

Particulate Matter

Should Comply

Pyrogens

As per specification

Sterility

Should comply

Assay

450000.0 to 550000.0 Units

5. METHOD OF ANALYSIS

Method Of Analysis of Finish Product :

Description:
White lyophilized cake filled in a 5ml amber colour glass vial labeled, stoppered and sealed with
blue colour flip off seal, such one vial packed in PVC tray with cover, such one tray along with
packing insert is packed in a carton.
Identification:
The retention time of test chromatogram is same as of standard chromatogram.
pH:
(Limit 5.0 To 7.0)
Lift electrode holding clamp at the appropriate height on the rod of the electrode stand and clean the
electrode with a jet of distilled water and wipe. Dry with tissue paper. Dip the electrode in the sample
solution & allow sufficient time for electrode to attain temperature.
Set 'selector switch ' to pH position and wait for 30 sec .The value of the pH sample will be displayed on
the 'Read Out' Note the reading. The reading of pH value should be stable within 0.01 for repetitive
operations if it is not then check the sample solution for freshness and electrodes and instrument for
proper functioning.
Set the 'Selector switch ' to stand by position .Raise the electrode remove the container of sample. solution
clean the electrode with distilled water and wipe, it with tissue paper and dip the electrode in distilled
water.
Determination of Water:
(NMT 5.0% w/w)
Titrimetric Method (Step-1):
Primary Standardization of the reagent: Place about 36ml of dehydrated methanol in the titration
vessel and add sufficient Karl-Fischer Reagent to give the characteristics end-point. Add quickly about
30 mg of Water, accurately weight by difference and titrate to the end-point.
STEP-2: Transfer sufficient quantity of sample accurately weight to the titration vessels containing the
water free methanol from step-(1), and titrate with Karl-Fischer reagent to the electrometric end point.
Care should be taken during weighing that the material is transferred immediately to avoid any exposure
to atmospheric humidity. Read and the percentage of the water from the instrument display or calculate
as follow:
vol. of KF reagent x KF factor x 100
----------------------------------------------=
% w/w
Wt. of sample (in mg)
Particulate Matter:
The Particulate matter analysis is done by the help of Particle Measuring Systems.
The limit of particle size is given below:
110m particle size < 6000 per container
225m particle size < 600 per container
Sterility: (Membrane Filtration Method)
1- Take 20 vials of Polymyxin B sulphate inj and dilute each container with sterile peptone water and
transfer aseptically as below:
Remove the lid of cup, pour the sample solution in to the cup and closed the lip using a

membrane filters with a pore size of 0.45m & 47mm.

2-

Filter the solution under vacuum pressure or reduced pressure.

3-

Wash the membrane with three successive quantities of 100 ml 0.1% sterile peptone water.

4-

Aseptically remove the membrane from the filter, cut it in to equal halves.

5-

Immerse one half the membrane in sterile 100 ml Fluid Thioglycoiiate Medium tube &
another in sterile 100 ml Soyabean Casein Digest Medium.
Note- Positive control do not performed in sterility testing area.

6-

Incubate Soyabean Casein Digest Medium at 20C to 22C and Fluid Thioglycoiiate
Medium tubes at 30 C to 35C for 14 days

7-

Observed the tubes for turbidity /growth after 24 hours. Note the observation. No growth
/turbidity should be observed in sample and negative control media tubes.

Observation and interpretation:

1- If no evidence of microbial growth is found, the prepation being examined passes the test of
sterility.
2- If evidence of microbial growth is found, reserve the container showing this and unless it is
demonstrated by any other means that their presence is due to causes unrelated to preparation
being examined and hence the tests for sterility is invalid. Performed a retest using same quantity /
number of sample as in the original test.
3- If no evidence of microbial growth is found in found in retest sample, the prepation being
examined passes the test of sterility.
4- If no evidence of microbial growth is found in retest sample, isolate &identify the organism. If
they are not readily distinguishable from those in original sample, the prepation being examined
fails the test of sterility.
5- If they are not readily distinguishable from those in original contaminant, perform a second
results twice number/ quantity of samples.
6- If no evidence of microbial growth is found in second retest sample, being examined passes
the test of sterility& if evidence of microbial growth is found in second retest sample examined
fails the test of sterility.

Pyrogen :
If intended for use in the manufacture of parenteral dosage form without a further appropriate procedure
for the removal of pyrogens, it complies with the test for pyrogens inject, per kilogram of the rabbits
mass, 1ml of a solution in water for injections R containing 1.5mg of the substance to be examined per
millilitre.

ASSAY: (BY HPLC) :

(Limit: 450000 units to 550000 units )

Requirement:
Apparatus:
A suitable HPLC capable of performing isocratic, UV-VIS detector, sample injector, controller and
integrator or equivalent software. The system is equipped with C-18, 4.6X250 mm 5 column
(Symmetry ODS is suitable).
Reagent: Anhydrous sodium sulphate, Acetonitrile (HPLC grade) and Water (Milli-Q).
Buffer preparation : Dissolve 4.46gm of anhydrous Sodium sulphate in 900ml water and adjust to pH
2.3 with dilute phosphoric acid and dilute to 1000ml with water.
Mobile Phase:
Mix 750 volume of Buffer and 250 volume of Acetonitrile make adjustment if necessary. Filter through
0.45m or finer porosity membrane filter.
Diluent :
Acetonitrile : Water - 20 : 80
Preparation of standard solution:
Weight accurately 50 mg Polymyxin B sulphate in to 50ml volumetric flask, and dilute with diluent up to
50ml. Obtained concentration of standard of 1000 mcg/ml.
Preparation of test solution:
Take weight of powder eq. to 50 mg Polymyxin B sulphate in to 50ml volumetric flask, and dilute with
diluent up to 50ml.Obtained concentration of sample of 1000mcg/ml .
HPLC Condition:
Instrument
: HPLC equipped with UV-Detector and suitable software
Column

: C-18, 4.6X250 mm 5 column (Symmetry ODS is suitable).

Wave length

: 215 nm

Flow rate

: 1.0 ml/minute

Injection volume : 20 l
Procedure: Inject diluent as blank. Inject separately standard and test preparation, run the
chromatograph, record the chromatogram, measure the maximum response for standard and test
Calculate the content of Polymyxin B sulphate as follow:
Calculation:
Area of Test x Conc. of Std (mcg)
----------------------------------------------- - x Potency of Std on as is basis x Avg. weight
Area of Std. x Conc. of Test (mcg)

6.

SAMPLES OF LABELS & CARTONS

7. SAMPLES WITH COA

SAMARTH LIFE SCIENCES PVT. LTD.

NANGAL UPERLA, SWARGHAT ROAD,
NALAGARH, DIST. SOLAN, H.P. 174101
QUALITY CONTROL DEPARTMENT
(Under drugs and cosmetic act, 1940 and the Rules there under)

NAME OF THE PRODUCT

GENERIC NAME
BATCH NO.
MFG. DATE
EXP. DATE
BATCH SIZE
DATE OF ANALYSIS

:
:
:
:
:
:
:

CERTIFICATE OF ANALYSIS
Poly B Injection
Polymyxin B Sulphate Injection
2PLB12
May 2013
April 2016
19127 Nos.
21/05/2013

TESTS

SPECIFICATION

RESULTS

Description

White lyophilizedcake filled in 5 ml

amber colour glass vial, labelled,
stoppered and sealed with blue colour
flip off seal, such one vial packed in
PVC tray with cover, such one tray
along with packing insert is packed in
a carton.

Identification

As per specification

White lyophilizedcake filled in 5

ml amber colour glass vial,
labelled, stoppered and sealed
with blue colour flip off seal,
such one vial packed in PVC tray
with cover, such one tray along
with packing insert is packed in a
carton.
Complies

pH

5.0 to 7.0

6.10

Average Filled Weight

As per specification

65.4 mg

Water

NMT 5.0% w/w

3.5% w/w

Particulate Matter

Should Comply

Complies

Pyrogens

As per specification

Complies

Sterlity

Should comply

Complies

*Assay
Each Vial Contains

Stated Amount (Units /

vial)

Polymyxin B Sulphate 500000.0 units

Esti. Amount (Units /

Vial)

Limits

505363.11 Units

450000.0 to 550000.0
units

*This indicate that this test done by public Lab report No. D/13/1206/029
Result: Sample Complies under above prescribe as per IHS specification.
Date: 15/06/2013

ANALYST

Q.C.INCHARGE

8. ANALYTICAL REPORT

9. STABILITY DATA

STABILITY DATA
Protocol for Stability Study, Real Time Stability study data and Accelerated Stability Study Data for three
batches have been attached.

PROTOCOL FOR STABILITY DATA

POLY-B (Polymyxin B Sulphate Injection 500000 IU)
Samples :
Three batches of POLY-B (Polymyxin B Sulphate Injection 500000 IU) are being tested:.
Batch No.

Mfg. Date

Exp. Date

Storage Time

2PLB
2PLB
2PLB
Composition:
Each vial contains :
Polymyxin B Sulphate BP .......................... 500000 IU
Water for Injections IP ................................ q.s.

Packaging and Packaging Material :

5 mL National Amber, Moulded, Type 1 glass vial with light blue coloured flip-off.
Storage Conditions:
The tested batches were stored at the following storage conditions:
Real Time conditions

30C 2C / 75 % 5%

Accelerated conditions

40C 2C / 75 % 5%

The Monitored Parameters:

The following parameters of POLY-B (Polymyxin B Sulphate Injection 500000 IU) were monitored:
1. Description
2. pH
3. Water
4. Particulate Matter
5. Bacterial Endotoxin
6. Sterility
7. Assay

Requirements:
1. Description : White lyophilized cake filled in 5 ml amber colour glass vial, labeled, stoppered and sealed
with blue colour flip off seal, such one vial packed in PVC tray with cover, such one tray along with packing
insert is packed in a carton.

2. pH : 5.0 to 7.0

3. Water : NMT 5.0% w/w

4. Particulate Matter : Should be free from particulate matter.
5. Sterility : Should be sterile.
6. Assay : 450000.0 to 550000 Units
Results:
The results are shown in the table for Real time studies and Accelerated studies.
Conclusions:
The present stability testing results show that POLY-B (Polymyxin B Sulphate Injection 500000 IU)
meets all the requirements of the prescribed specifications.
Life and Storage Recommendation:
According to the present results of Stability testing of POLY-B (Polymyxin B Sulphate Injection 500000
IU), shelf life of 24 months can be proposed.

REAL TIME STABILITY

STUDY DATA OF THREE BATCHES

ACCELERATED STABILITY
STUDY DATA OF THREE BATCHES