Annexure No-3

OM BIOMEDIC PVT.LTD. HARIDWAR OQA/SOP/030/A03-03

44 FINISH PRODUCT SPECIFICATION
SOP No. OQA/SOP/030° | REVISION No.

TITLE: CLOTRIMAZOLE, MENTHOL, ICHTHAMMOL, BORIC ACID & ZINC OXIDE CREAM

Market: Export Reference IHS

sass provaet Supersedes No. Effective Date Review Date Page No.
Specification No. 4
ere legge cep cn Nil 07/04/2024 06/04/2026 | Page lof 1

S. No TEST SPECIFICATION

1. “Description A yellow semi solid mass filled in Lami tube sealed properly.
25 Identification Should give positive result for Clotrimazole, Menthol, Ichthammol,
Boric Acid & Zinc Oxide.
3: Average fill weight 20.0 gm + 9.0 %

4. Uniformity of fill weight + 9.0 % of labeled amount.

5. Assay
Fach gm contains:
Clotrimazole BP 0.5% w/w 0.45% w/w to 0.55 % w/w (90.0% to 110.0%)
Menthol 1.0% w/w 0.90 % w/w to 1.10 % w/w (90.0% to 110.0%)
Ichthammol BP 0.20% w/w > 0.18% w/w to 0.22% w/w (90.0% to 110.0%)
Boric Acid BP — 1.00% w/w 0.90 % w/w to 1.10 % w/w (90.0% to 110.0%)
Zinc Oxide BP 5.00% w/w 4.5 % wi/w to 5.5 % w/w (90.0% to 110.0%)
6. | Microbiological Test
TAMC 10° cfu/g
TYMC 10' cfu/g
Staphylococcus aureus Should be absent/g
Pseudomonas aeruginosa Should be absent/g

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Date 0 &loY}r0}4 \ o6loy}r0r4t 7k

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Name & Designation Nishtha Kansal Ranveer Singh “S. kshay Negi /.§
(Executive QC) (Sr. Executive QC) rd ev
Format No.: OQA/SOP/030/A03-03
AA) OM BIOMEDIC PVT.LTD. HARIDWAR anasto
4A FINISH PRODUCT TESTING PROCEDURE
SOP No. OQA/SOP/030 [_ REVISION No. 03
TITLE: CLOTRIMAZOLE, MENTHOL, ICHTHAMMOL, BORIC ACID & ZINC OXIDE CREAM
Market: EXPORT
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STP No. Ref. STS No. No. Date Date Reference | Page No.
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LABORATORY SAFETY:
1.0 Good Laboratory practice should be followed while handling & preparing all reagents.
2.0 Mouth pipetting is not allowed, pipette bulbs should be used.
3.0 Always add acid to water. Care should be taken in the preparation.
Strong acid & alkalis should be prepared under a suitable fume hood. Safety glasses & mask must be worn while
handling.

1.0 Description:
Procedure: Take 5 g cream in Petri dish and physically observe colour and appearance of the sample.

2.0 Identification:
Should give positive result for Cleuinazcle. Menthol, Ichthammol, and Boric Acid & Zine Oxide.

3.0 Average weight:

Equipment: A suitable analytical balance.
Procedure:
Weigh 10 tubes selected at random and weigh after clean the tube calculate the average weight by following formula.

rs Weight of 10 Net weight of tube

Average fill weight =
10
.

4.0 Uniformity of weight:

Equipment: A Suitable analytical balance.
Procedure: Select a sample of 10 filled tubes & remove any labeling that might be altered in weight, while
removing the contents of containers. Clean & dry the outer surfaces of the tube & weigh each tube. Remove
quantitatively the contents from each tube. If necessary, cut open the container & wash each empty container with
a suitable solvent, taking care to ensure that the closure & other parts of the container are retained. Dry & again
weigh each empty container together with its parts which may have been removed. The difference between the two
weights is the net weight of the contents of the container.
The average net weight of the contents of the 10 containers is not less than the labelled amount and the net weight
of the content of any single container is not less than 91% and not more than 109% of the labelled amount where
the labelled amount is 50 g or less.

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Name & -. ‘Nishtha Kansal Ranveer Singh
Designation (Executive QC) (Sr. Executive QC)
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Format No.: OQA/SOP/030/A08-03
AA) OM BIOMEDIC PVT.LTD. HARIDWAR CBASnEee
44 FINISH PRODUCT TESTING PROCEDURE
SOP NeNo. OQA/SOP/030 | REVISION No. 03
TITLE: CLOTRIMAZOLE, MENTHOL, ICHTHAMMOL, BORIC ACID & ZINC OXIDE CREAM
Market: EXPORT
Supersedes Effective Review
STP No. Ref. STS No. No. Date Date Reference | Page No.

OM/FP/STP/CMI | OM/FP/STS/CMIB , 2 of 8
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Minimum weight = Minimum weight/ Nominal weight x 100

Maximum weight = Maximum weight/ Nominal weight x 100

5.0 Assay
For Clotrimazole
Chromatographic Condition
Column : 200 mm x 4.6 mm x Sum
Injection Volume : 20 ul
Flow Rate : 1.5 ml/min
Wavelength > 215 nm

Mobile Phase
30 volumes of 0.02M miguneolone acid and 70 volumes of methanol, the pH of the mixture being adjusted to 7.5
with a 10% v/v solution of triethylamine in methanol.

(1) Extract a quantity of the cream containing 20 mg of Clotrimazole by warming with20 mL of methanol in a water
bath at 500 for 5 minutes, shaking occasionally. Remove from the water bath, shake the mixture vigorously while
cooling to room temperature, cool in ice for 15 minutes, centrifuge for 5 minutes and decant the supernatant liquid.
Repeat the extraction with two further 20-mL quantities of methanol. To the combined methanol extracts add 10 mL
of methanol and dilute to 100 mL with water. Cool in ice and filter through a glass micro fibre filter paper (Whatman
GF/C is suitable). Dilute 2 volume of the filtrate to 10 volumes with a mixture of 3 volumes of water and 7 volumes
of methanol.
(2) Dissolve 20 mg of clotrimazole BPCRS i in 10 mL of methanol, add sufficient water to produce 100 mL and dilute
2 volume of the resulting solution to 10 volumes with a mixture of 3 volumes of water and 7 volumes of methanol.

System Suitability
The column efficiency, determined using the peak in the oie ciara obtained with solution (2), should be at least
9000 theoretical plates per metre.

Calculate the content of Clotriamzole by following formula.

Prepared By: ked By

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Date o6\oy\ 194 06[04 | 024

Name &” Nishtha Kansal Ranveer Singh
Designation (Executive QC) (Sr. Executive QC)
Format No.: OQA/SOP/030/A08-03
 No-8
> OM BIOMEDIC PVT.LTD. HARIDWAR ouOesE
44 FINISH PRODUCT TESTING PROCEDURE
SOP No. OQA/SOP/030: |. REVISION No. 03
of

TITLE: CLOTRIMAZOLE, MENTHOL, ICHTHAMMOL, BORIC ACID & ZINC OXIDE CREAM
Market: EXPORT
ee Supersedes Effective Review
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AY We i 2 100 10 P
x x x x x x 100

Where,
AT ="Average area of the sample solution,
AS = Average area of the standard solution,
Ws = Weight of working standard in mg,
WT = Weight of the sample taken in the test in mg,
P = Potency of working standard (as such basis),

For Menthol
Chromatographic conditions:
Column : HP-5 (30mx0.320 mm x 0.25 um
Column Oven Temperature: 85° C hold for 2 min and then increase to 200°C at the rate 10 C/min and hold for 8 min.
Detector : FID
Detector Temperature $285" Co's
Injection port temperature : 250°C
Injection Volume > 4um
Carrier gas : Nitrogen /helium
Carrier gas Flow ~ + 8.0 ml/min
Diluegt : MDC
Run Time : About 22 minutes
Split = 1310

Standard Preparation: Weigh and transfer about 20 mg of menthol working standard into a 100 ml volumetric
flask. Add 70 ml of diluent and shake well. and dilute upto the mark with the diluent. Further dilute 5 ml of this
solution to 50 ml, add 10 ml of methanol and then make up volume with MDC.

Sample Preparation: Weigh and transfer 5 gm of sample into a 50 ml volumetric flask. Add 10 ml of methanol and
Sonic&te for 5 mins. Add 20 ml of diluent and vortex the sample. Dilute up to the mark with diluent and take MDOC
Layer (Organic layer) and fill in 2ml vial

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Format No.: OQA/SOP/030/A08-03
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AA) OM BIOMEDIC PVT.LTD. HARIDWAR Sa SRAB LE
44 FINISH PRODUCT TESTING PROCEDURE
SOP No. OQA/SOP/030 | REVISION No. 03

TITLE: CLOTRIMAZOLE, MENTHOL, ICHTHAMMOL, BORIC ACID & ZINC OXIDE CREAM
Market: EXPORT
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Calculation:

Avg Areaofspl Std. Wt. Dilution Potency of Standard CD

xX xX x X 1
Avg Area of Std. Dilution Sample Wt. 100:

For Zinc Oxide

Procedure: ;
Dissolve 0.150 g in 10 mL of dilute acetic acid. Carry out the complex metric titration of zinc .
1 mL of 0.1 M sodium edetate is equivalent to 8.14 mg of ZnO.
s

Calculation:
Volumes Consumed X: Morality X Equivalent Factor X100

Weight of Sample X Stated Morality

For Boric Acid

Procedure:
Dissolve 1.000 g with heating in 100 mL of water R containing 15 g of Mannitol. Titrate with 1 M sodium
hydroxide, using 0.5 mL of phenolphthalein solution R as indicator, until a pink colour is obtained.
1 mL of | M sodium hydroxide is equiyalent to 61.8 mg of H3BO3.

Calculation:
Volumes Consumed X Morality X Equivalent Factor X100

Weight of Sample X Stated Morality

.
For Ichthammol
Sample: Transfer a portion of the Ointment, equivalent to 2 g of Ichthammol, to a 250-mL beaker, and add 70 mL of
boiling water. Mix with a glass rod, and heat on a steam bath with frequent agitation for 10 min. Cover with a watch
glass without removing the stirring rod, and allow to stand at room temperature for 15-20 min. Place in a refrigerator

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Date & 6(0Y)r0ry 06/0Y/r924

Name & _Nishtha.Kansal | Ranveer Singh
Designation (Executive QC) (Sr. Executive QC)
Format No.. OQA/SOP/030/A08-03
 OM BIOMEDIC PVT.LTD. HARIDWAR Puyo ears
4A FINISH PRODUCT TESTING PROCEDURE
SOP No, OQA/SOP/030 | REVISION No. 03
TITLE: CLOTRIMAZOLE, MENTHOL, ICHTHAMMOL, BORIC ACID & ZINC OXIDE CREAM
Market: EXPORT
Supersedes Effective Review
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to cause the upper layer to congeal, form a second opening through the congealed layer with the glass rod, and
transfer the dark-colored aqueous extract to a funnel containing a pledget of cotton, collecting the filtrate in a 500-mL
volumetric flask. Repeat the extraction of the portion of the Ointment several times in the same manner until the
aqueous extract is practically colorless, passing each extract through the same cotton filter into the flask containing
the main extract. Dilute with water to volume, and mix.
Titrimetric system :
Mode: Residual titration
Titrant: 0.05 N sulfuric acid VS
Back-titrant: 0.05 N sodium hydroxide VS
Endpoint detection: Colorimetric
Analysis: Transfer 100.0 mL of the Sample to a suitable distillation flask. add 3 g of paraffin, and add 20 mL of
sodium hydroxide solution (4 in 10). Connect the flask to a condenser by means of a spray trap, and immerse the
lower outlet tube of the condenser in 30.0 mL of Titrant. Distil slowly, collect about 50 mL of distillate, and then
titrate the excess sulfuric acid with Back-titrant, using methyl red TS as the indicator. Perform a blank determination,
and make any necessary correction.
Each mL of Titrant is equivalent to 0.8515 mg of ammonia (NH3).

6.0 Microbiological Test

Sample preparation:
Water soluble products:
Dissolve 10 gm or 10 ml of the product in 90 ml sterile phosphate buffer pH 7.2 / sterile buffered sodium chloride
peptone solution pH 7.0/ Soyabean casein digest medium (usually 1 in 10 dilutions). If required, make up the final
volume up to 100 ml. Mix the sample/content properly.
Water insoluble Non Fatty products:
Dissolve 10 gm or 10 ml of the product in 90 ml Sterile Phosphate buffer pH 7.2 / Sterile Buffered Sodium Chloride
Peptone Solution pH 7.0 / Soyabean Casein Digest Medium with Polysorbate 20/80 (4.0%) (Usually 1 in 10
dilutions). If required, make up the final volume up to 100ml. Mix the sample/content properly.
Fatty Products:
Homogenize 10 gm or 10 ml of the product in 90 ml Sterile Phosphate buffer pH 7.2 / Sterile Buffered Sodium
Chloride Peptone Solution pH 7.0 / Soyabean Casein Digest Medium with Polysorbate 20/80 (4.0%) and mix well
(usually 1 in 10 dilution). If necessary Sterile Buffered Sodium Chloride Peptone Solution pH 7.0 / Soyabean Casein
Digest Medium subject the contents of the tube for warming to a temperature not exceeding 45°C. Make up the final
volume up to 100 ml.

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Date o6[o4lror4 66 [O4|L024

Name & Nishtha Kansal Ranveer Singh
Designation (Executive QC) (Sr. Executive QC)
Format No.: OQA/SOP/030/A08-03
 OM BIOMEDIC PVT.LTD. HARIDWAR meagan
4A FINISH PRODUCT TESTING PROCEDURE
SOP No. OQA/SOP/030. - |. REVISION No. 03

TITLE: CLOTRIMAZOLE, MENTHOL, ICHTHAMMOL, BORIC ACID & ZINC OXIDE CREAM
Market: EXPORT
Supersedes Effective Review
STP No. Ref. STS No. No. Date Date Reference | Page No.
OM/FP/STP/CMI_ | OM/FP/STS/CMIB ‘ 6 of 8
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Test for Total Aerobic Microbial count (TAMC) and Total Yeast and Mould count (TYMC):
Pour-plate method:
Aseptically transfer 1ml of pretreated sample into four different Petri dish (2 of SCDA and 2 of SDA/SCA), which is
previously labeled and kept under Laminar Air Flow.
Pour approximately 15-20 ml of SCDA (in 2 plates) and SDA/SCA (in 2 plates) media into the respective plates and
rotate"the plates clockwise and anti clockwise to mix the contents and allow to solidify.
Incubate the SCDA plates at 30-35°C for 3-5 days & SDA plates at 20—25°C for 5-7 days.
Positive Control:
Add known volume of cells (10-100 CFU/ml) from one of the mention organisms S. aureus ATCC
6538, Pseudomonas aeruginosa ATCC 9027 in each plate & pour 15-20 ml SCDA. Gently rotate the plates for the
even distribution of cultures and allow solidifying. Incubate the plates of bacterial cultures at 30-35°C for 2-3 days.
Add known volume of cells 10-100 CFU/ml of Candida albicans ATCC 1023 and Aspergillus brasiliensis ATCC
16404 in each plate & pour 15-20 ml SDA. Gently rotate the plates for the even distribution of cultures and allow
solidifying. Incubate the plates at 20—25°C for 3-5 days.
Negative Control:
Pour the SCDA / SDA/SCA Media in er rrvee and labeled plates without sample and allow solidifying.
Interpretation:
Count the colonies produced and spark the results as CFU/total volume of sample tested. Negative Control plate
must not show any CFU. Positive Control plate must show luxuriant growth within 48-72 hours of incubation for
bacteria and 72-120 hours for fungi. The mean of the two plates calculated with dilution factor and quantity of
sample finally expressed as CFU/ gm of Sample.
CFU/gm/ml = Average No. of Colonies X Dilution Factor
eis Volume of sample tested.
The total aerobic microbial count (TAMC) is considered to be equal to the number of CFU found using soybean
casein digest agar (SCDA)); if colonies of fungi are detected on this medium, they are counted as part of the TAMC,
The total combined yeasts/mould count (TYMC) is considered to be equal to the number of CFU found using
Sabouraud dextrose agar (SDA); if colonies of bacteria are detected on this medium, they are counted as part of the
TYMC. When the TYMC is expected to exceed the acceptance criterion due to the bacterial growth, Sabouraud
dextrose agar containing antibiotics may be used.

Membrane Filtration Method:

Aseptically transfer 1ml of pretreated sample through 0.45 um pore size membrane having 47mm diameter. Rinse the
membrane with minimum three 100ml portions of 0.1% peptone water. Number of rinses can be increased as per the
validation report for the product up to 2'to 3 times.

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Date © bl oy]10)4 06/04 /r0ry

Name & Nishtha Kansal Ranveer Singh
Designation (Executive QC) (Sr. Executive QC)
Format No.: OQA/SOP/030/A08-03
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Fy OM BIOMEDIC PVT.LTD. HARIDWAR earn a

4 f FINISH PRODUCT TESTING PROCEDURE
SOP No. OQA/SOP/030 | REVISION No. 03

TITLE: CLOTRIMAZOLE, MENTHOL, ICHTHAMMOL, BORIC ACID & ZINC OXIDE CREAM
Market: EXPORT
Supersedes Effective Review
STP No. Ref. STS No. No. Date Date Reference | Page No.

OM/FP/STP/CMI_ | OM/FP/STS/CMIB : 7 of 8
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With the help.of sterile forceps transfer the membrane to the surface of pre-incubated Soybean Casein Digest Agar
(SCDA) plate for determination of total aerobic microbial count (TAMC).
Repeat as mentioned above and transfer the membrane to the surface of pre-incubated Sabouraud Dextrose Agar
(SDA) or Sabouraud Chloramphenicol agar (SCA) for the determination of total combined yeast and mold (TYMC).
Incubate the SCDA plate at 30-35°C for 3 to 5 days and SDA/SCA plate at 20-25°C for 5 to 7 days.
Positive Control ;
Aseptically filter 100 ml of BSCP/ sterile 0.1% of Peptone water, which previously inoculated with one of mention
organism 10 to 100 cells of S. aureus ATCC 6538, Pseudomonas aeruginosa ATCC 9027 and place the filter paper
on pre incubated SCDA plates. Incubate the plates of bacterial cultures at 30—35°C for 3 days. Aseptically filter 100
ml of BSCP/ Sterile 0.1% of Peptone water, which is previously, inoculated with 10 to 100 cells of C.
albicans ATCC 10231 &Aspergillus brasiliensis ATCC 16404 & place the filter paper on pre incubated SDA/SCA
plates. Incubate the SDA/SCA plates at 20-25°C for 5 days.
Negative Control:
Aseptically filter 100 ml quantity of BSCP/ Sterile 0.1% Peptone Water by the same procedure followed for the
sample & incubate the SCDA plate at 30 -35°C for 3-5 days & SDA/SCA plate at 20-25°C for 5-7 days.
Interpretation:
Count the colonies produced and report the results as CFU/total volume of sample tested. Negative Control plate
must not show any CFU. Positive Control plate must show luxuriant growth within 48-72 hours of incubation for
bacteria and 72-120 hours for fungi. The mean of the two plates calculated with dilution factor and quantity of
sample finally expressed as CFU/ gm of Sample.
CFU/gm/ml = Average No. of Colonies X Dilution Factor
Volume of sample tested.

Tests for Pathogens:

Take 10gm/10ml of early prepared sample of product and inoculate it in to 90mlSoybean Casein Digest Medium
(SCDM). Incubate at 30-35°C for 18-24 hours.

Pseudomonas aeruginosa
Using Casein soyabean digest broth as a diluent make 1 in 10 dilution of more than | g of the product as mentioned
in Total aerobic viable count under Microbial contamination in nonsterile‘products and use 10 ml or the quantity
corresponding to 1 g or | ml of the product to inoculate a suitable amount (determined as under Validity of the Test
method) of Casein soyabean digest broth incubate at 30° to 35° for 18 to 24 hours Subculture on a plate of Cetrimide
agar (Medium 13) and incubate at 30“ to 35° for 18 to 72 hours.
A greenish color colony indicates the possibility of presence of Pseudomonas aeruginosa. This should be confirmed
by identification tests.

= Prepared By opened By
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Date o6l04/) 024 0 6(04/10 14

Name & Nishtha Kansal -Ranveer Singh
Designation (Executive QC) (Sr. Executive QC)
Format No.: OQA/SOP/030/A08-03
AA) OM BIOMEDIC PVT.LTD. HARIDWAR Bronte
4A FINISH PRODUCT TESTING PROCEDURE
SOP No. OQA/SOP/030 | REVISION No. 03

TITLE: CLOTRIMAZOLE, MENTHOL, ICHTHAMMOL, BORIC ACID & ZINC OXIDE CREAM
Market: EXPORT
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If there is no growth of such type of colonies, or identification tests are negative it indicates absence of P aeruginosa
and the product passes the test. :
Staphylococcus aureus ,
Using Casein soyabean digest broth as a diluent make 1 in 10 dilution of more than 1 g of the product as mentioned
in Total aerobic viable count under Microbial contamination in Non sterile products and use 10 ml pr the quantity,
corresponding to.d g or 1 ml of the product to inoculate a suitable! amount (determined as under Validity of the Test
method) .of Casein soyabean digest broth incubate at 30° to 35° for 18to 24hours.
Sub-culture on a plate of Mannitol salt agar (Medium-14) incubate at- 30° to 35° for 18 to-72 hours. Yellow to white
colonies with yellow zones indicate the possibility of presence of S. aureus; This should be confirmed b
identification tests.
If there is no growth of such type of colonies, or the identification tests are negative it indicates absence of
S. aureus and the product passes the test.

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Date poy] or o6ley/r0 | fa( o¢foy/t02) «
Name & Nishtha Kansal Ranveer Singh Akshay Negi
Designation (Executive QC) _ . (Sr. Executive QC) <2 SS
Format No.: OQA/SOP/030/A08-03