FEROZSONS LABORATORIES LIMITED
WORK INSTRUCTIONS
W.I. No fQ-280-00
Effective Date: February 15, 2016
Review Date: February 15, 2019
Page No. Page 1 of 6
.
TESTING INSTRUCTIONS OF NEOMYCIN SULFATE
Title Signature Date
Written by Deputy Manager Quality Control
Reviewed by Manager Quality Control
Reviewed by Manager Quality Assurance
Approved by Director Quality Operations
Reference: United State Pharmacopoeia
1. APPEARANCE: White to slightly yellow powder, or cryodesiccated
solid. Is odorless or practically so and is hygroscopic.
Its solutions are dextrorotatory.
2. SOLUBILITY: Freely soluble in water; very slightly soluble in
alcohol; insoluble in acetone, in chloroform, and in
ether.
3. IDENTIFICATION:
Solution (1 in 20) responds to the tests for Sulfate.
Procedure:
When treated with 3 N hydrochloric acid, sulfites and bisulfites yield sulfur
dioxide, which blackens filter paper moistened with mercurous nitrate
4. pH:
All or part of the information contained in this document should be treated as the confidential property of Ferozsons
Laboratories Limited. It cannot be published or divulged for whatever purpose to unauthorized persons, or 3rd parties, in
any form without the prior written permission from DQO Ferozsons Laboratories Limited.
COPY NO.
FEROZSONS LABORATORIES LIMITED
WORK INSTRUCTIONS
W.I. No fQ-280-00
Effective Date: February 15, 2016
Review Date: February 15, 2019
Page No. Page 2 of 6
.
TESTING INSTRUCTIONS OF NEOMYCIN SULFATE
In a solution containing 33 mg of Neomycin per mL
LIMIT: between 5.0 and 7.5
5. LOSS ON DRYING:
Weigh accurately about 100 mg of the substances in a tarred watch glass. Before
noting weighing spread the sample in watch glass evenly. Place watch glass in
oven and dry the sample at 800C + 20C till constant weight (3 hours under
vacuum). Allow the sample to come to room temperature by keeping it in
Desiccator. Weigh the sample and calculate the percentage by following formula.
W1 - W 2
% w/w = --------------- X 100
W1
W1 = Weight of the sample before drying
W2 = Weight of the sample after drying
LIMIT: NMT 8.0% of its weight
6. SULPHATED ASH:
Ignite a suitable crucible (for example, silica, platinum, porcelain or quartz) at 600
+ 50 °C for 30 min, allow to cool in a desiccator over silica gel or other suitable
desiccant and weigh. Place the prescribed amount of the substance to be
examined in the crucible and weigh. Moisten the substance to be examined with
Reviewed by: MQA Sign: Date
All or part of the information contained in this document should be treated as the confidential property of Ferozsons
Laboratories Limited. It cannot be published or divulged for whatever purpose to unauthorized persons, or 3rd parties, in
any form without the prior written permission from DQO Ferozsons Laboratories Limited.
COPY NO.
FEROZSONS LABORATORIES LIMITED
WORK INSTRUCTIONS
W.I. No fQ-280-00
Effective Date: February 15, 2016
Review Date: February 15, 2019
Page No. Page 3 of 6
.
TESTING INSTRUCTIONS OF NEOMYCIN SULFATE
a small amount of sulfuric acid (usually 1 ml) and heat gently at as low
temperature as practicable until the sample is thoroughly charred. After cooling
moisten the residue with a small amount of sulfuric acid (usually 1 ml), heat
gently until white fumes are no longer evolved and ignite at 600 + 50 °C until the
residue is completely incinerated. Ensure that flames are not produced at any
time during the procedure. Allow the crucible to cool in a desiccator over silica gel
or other suitable desiccant, weigh it again and calculate the percentage of
residue. If the amount of the residue so obtained exceeds the prescribed limit,
repeat the moistening with sulfuric acid R and ignition, as previously, for 30 min
periods until 2 consecutive weighing do not differ by more than 0.5 mg or until the
percentage of residue complies with the prescribed limit. The amount of
substance used for the test (usually 1-2 g) is chosen so that at the prescribed
limit the mass of the residue (usually about 1 mg) can be measured with
sufficient accuracy.
CALCULATION:
W2
% age = ---------------- X 100
W1
W1 = weight of substance taken
W2 = Weight of substance after ignition
LIMIT: Maximum 1.0 %
7. MICROBIAL ASSAY:
Reviewed by: MQA Sign: Date
All or part of the information contained in this document should be treated as the confidential property of Ferozsons
Laboratories Limited. It cannot be published or divulged for whatever purpose to unauthorized persons, or 3rd parties, in
any form without the prior written permission from DQO Ferozsons Laboratories Limited.
COPY NO.
FEROZSONS LABORATORIES LIMITED
WORK INSTRUCTIONS
W.I. No fQ-280-00
Effective Date: February 15, 2016
Review Date: February 15, 2019
Page No. Page 4 of 6
.
TESTING INSTRUCTIONS OF NEOMYCIN SULFATE
Microbial Assay is performed by Well Diffusion method, 8 wells are bored in the
agar surface with the help of sterile borer.
Procedure – Method:
After preparing microbial suspension (see WI No. M-58), dip sterile swab to the
bacterial suspension tube.
Inoculate the plates by dipping a sterile swab into the inoculum, remove excess
inoculum by pressing and rotating the swab firmly against the side of the tube
above the level of the liquid.
Streak the swab all over the surface of the Mueller Hinton Agar Medium (M – H
Agar) three times for making complete lawn of bacterial suspension, streaking
the plate through an angle of 60° after each streaking stroke, and finally, pass the
swab round the edge of the agar surface (see Fig. 1 – given below).
Leave the inoculum to adjust & dry for 3 – 5 minutes (not more than 15 minutes)
at room temperature with the lid closed under the L.F.C. / B.S.C.
After adjustment of inoculum with the agar, make 8 wells / holes with the help of
sterile borer of 5 – 8 mm in diameter evenly for sample and standard antibiotic
solutions.
The wells / holes are marked as Standard High (SH), Standard Low (SL), Sample
High (H) & Sample (L) twice in opposite directions (see Fig. 2).
0.1 mL of standard & sample solution is poured in their respective wells/holes.
The plate is left for 1 – 4 hours at room temperature as a period of pre-incubation
diffusion.
Incubate the petri plate containing standard and sample at the 30 – 35 °C for 16
– 24 hours (overnight) within 30 minutes after applying discs.
After incubation, observe the zone of inhibition around the sample and standard
antibiotic discs, and measure them with Vernier caliper or with measuring scale
and report zones in millimeter (mm) (see Fig. 3).
Reviewed by: MQA Sign: Date
All or part of the information contained in this document should be treated as the confidential property of Ferozsons
Laboratories Limited. It cannot be published or divulged for whatever purpose to unauthorized persons, or 3rd parties, in
any form without the prior written permission from DQO Ferozsons Laboratories Limited.
COPY NO.
FEROZSONS LABORATORIES LIMITED
WORK INSTRUCTIONS
W.I. No fQ-280-00
Effective Date: February 15, 2016
Review Date: February 15, 2019
Page No. Page 5 of 6
.
TESTING INSTRUCTIONS OF NEOMYCIN SULFATE
Calculate the % potency of the sample (in terms of the standard) from the
following equation.
Potency Calculation Formula for Neomycin
A = ½ {(H – L)} + {(SH – SL)}
B = ½ {(H – L)} – {(SH – SL)}
C = E/0.301
D = B/C
E = Antilog of D
%P = E X 100
Figure 1 – Template for making bacterial lawn (90 mm Diameter of Plate)
SH
L H
Reviewed by: MQA Sign: Date
All or part of the information contained in this document should be treated as the confidential property of Ferozsons
Laboratories Limited. It cannot be published or divulged for whatever purpose to unauthorized persons, or 3rd parties, in
any form without the prior written permission from DQO Ferozsons Laboratories Limited.
COPY NO.
FEROZSONS LABORATORIES LIMITED
WORK INSTRUCTIONS
W.I. No fQ-280-00
Effective Date: February 15, 2016
Review Date: February 15, 2019
Page No. Page 6 of 6
.
TESTING INSTRUCTIONS OF NEOMYCIN SULFATE
SL SL
H L
SH
Figure 2 – Template for making wells / holes for antibiotic standard and sample solutions
(150 mm Diameter of Plate)
Antibiotic Disc
Zone of Inhibition
Figure 3 – Template for measuring zone of inhibition in 2 way directions
Reviewed by: MQA Sign: Date
All or part of the information contained in this document should be treated as the confidential property of Ferozsons
Laboratories Limited. It cannot be published or divulged for whatever purpose to unauthorized persons, or 3rd parties, in
any form without the prior written permission from DQO Ferozsons Laboratories Limited.
COPY NO.
