PHARMA DEVILS

MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

DISINFECTANT VALIDATION
PROTOCOL & REPORT
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

1.0 DETAIL OF DISINFECTANT USE FOR VALIDATION:

NAME OF COMPOSITION BATCH. NO. CONCENTRATION
DISINFACTANT USE FOR
VALIDATION
ISO-PROPYL ALCOHOL Propan-2-ol L143841501 70% V/V

i) Chloroxylenol I.P. 4.8 % W/V

DETTOL ii) Terpineol B.P. 9.0 % V/V D4739 2.5% V/V
iii) Alcohol Absolute (Denatured) 13.1 % V/V

i) Chlorhexidine Gluconate Solution

I.P.1.5 % V/V
SAVLON KN4034 2.5% V/V
ii) Strong Cetrimide Solution equivalent to
Cetrimide 3.0 % W/V
i) Benzalkonium Chloride Solution
MICROLYSE MLE14058 1.5% V/V
I.P. 4.0 % W/V

i) Chloroxylenol I.P 2.4 % W/V

ii) Triclosan U.S.P. 0.5 % W/V
ACTALL AAL14004 2.5% V/V
iii)Terpineol B.P. 9 % W/V
iv)Iso-Propyl Alcohol I.P. 12% V/V

i) Chlorhexidine Gluconate
SAVINOX Solution I.P.1.5% V/V SNP14007 3.0% V/V
ii) Cetrimide I.P. 3.0% W/V
ENDOMAX i) Glutaraldehyde 2.45% W/V EMX14029 2.5% V/V
i) Silver Nitrate I.P. 0.01% W/V
SILVICIDE SCE5000 5.0% V/V
ii) Hydrogen Peroxide I.P. 10% W/V

FORMALDEHYDE i) Formaldehyde Solution (37-41% W/V) P12F100982 40% V/V

 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

2.0 PROTOCOL APPROVAL:

Particulars Name Signature Date

Prepared By
(Sr. Executive-QC)
Checked By
Head-QC
Approved By
Head-QA
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

TABLE OF CONTENTS:
S.No. TITLE PAGE No.
1.0 Detail of Disinfectant Use For Validation 2
2.0 Protocol Approval 3
3.0 Objective 5
4.0 Scope 5
5.0 Responsibility 5
6.0 Requirements 6
7.0 Reference 12
8.0 Evaluation of Results 12
9.0 Conclusion 12
10.0 Abbreviations 13
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

3.0 OBJECTIVE: To provide a procedure for disinfectant validation, for the surfaces and area sanitization of
controlled and clean rooms.

4.0 SCOPE: This Protocol is applicable for disinfectant validation to establish the “Minimum effective
concentration of disinfectant solution” and its effectiveness duration after application in
controlled and clean areas at ……………..

5.0 RESPONSIBILITIES:

DEPARTMENT RESPONSIBILITIES

 Responsible for follow the procedure.

 Responsible for preparation of validation protocol, summary report and supervise the
Quality Control
microbiology officer during the disinfectant validation activity.
 Responsible for review and technical correction of validation protocol.
 To Monitor all Validation Activities and ensuring the Validation as per the Protocol.
Quality
 To monitor Protocol completeness and Technical Accuracy
Assurance
 To Review the Data & Approve the Validated Procedure, if found satisfactory.
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

6.0 REQUIREMENTS: Before proceeding for validation following material are required.
6.1 CULTURES:
6.1.1 Staphylococcus aureus ATCC 6538
6.1.2 Pseudomonas aeruginosa ATCC 9027
6.1.3 Bacillus subtilis ATCC 6633
6.1.4 Escherichia coli ATCC 8739
6.1.5 Salmonella abony NCTC 6017
6.1.6 Shigella boydii ATCC 8700
6.1.7 Candida albicans ATCC 10231
6.1.8 Aspergillus brasiliensis ATCC 16404
6.1.9 Environmental isolate

6.2 DISINFECTANT:
6.2.1 Disinfectant for validation as per given table above on page no. 2.
6.2.2 Disinfectant validation study shall be carried out on below mentioned surfaces
Table-1
Sr. No. Surface Selection
1.0 Stainless steel
2.0 Plastic
3.0 Epoxy
4.0 Tiles
5.0 Glass
6.0 Aluminium
7.0 silicon

6.3 MEDIA AND OTHER REQUIREMENT:

6.3.1 Soyabean Casein Digest Agar (Hi- media MH290 or equivalent).
6.3.1.1 Preparation: Suspend 40 g in 1000 ml of purified water. Add 1 g per liter of
polysorbate 80 into the media. Sterilize the culture media by autoclaving at 15 psi,
121.1°C for 15 minutes. After sterilization pH should be 7.3 ± 0.2 at 25°C.
6.3.2 Buffered Sodium Chloride Peptone Solution pH 7.0 (Hi- media MH1275 or equivalent) as
Diluent.
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

6.3.2.1 Preparation: Suspend 14.64 g in 1000 ml of purified water. Add 1 g per liter of
polysorbate 80 into the media. Sterilize the culture media by autoclaving at 15 psi,
121.1°C for 15 minutes. After sterilization pH should be 7.0 ± 0.2 at 25°C.
6.3.3 Vortex Mixture
6.3.4 Sterile membrane filters
6.3.5 Auto pipette and sterile tips
6.3.6 Sterilized Petri plates
6.3.7 Forceps
6.3.8 Vaccum pump
6.3.9 Test Tubes Sterile.

7.0 VALIDATION TEST PROCEDURE FOR ESTABLISHMENT OF MINIMUM EFFECTIVE

CONCENTRATION OF DISINFECTANT:
7.1 DISINFECTANT VALIDATION USE DILUTION STUDY:
7.1.1 Culture concentration control
7.1.2 Inoculate 1 ml of culture suspension having more than 1,00,000 cfu in 10 ml of diluent.
7.1.3 Shake the tube gently and carry out serial dilutions in such a way to get less than 100 cfu count
on a filter paper.
7.1.4 Filter the dilution (giving less than 100 cfu) through 0.45 µ membrane filter by applying
Vaccum .Rinse twice the membrane filter with 100 ml diluent.
7.1.5 Aseptically transfer the membrane on the surface of sterile Soyabean casein Digest Agar with
Tween 80.
7.1.6 For bacterial culture incubate the plate at 30-35°C for 3-5 days and fungal culture at 20-25°C
for 5-7 days.
7.1.7 Count the actual number of inoculated cfu in the diluent as follow:
Actual No. of inoculated cfu = Observed cfu x Dilution factor.

7.2 MICROORGANISMS RECOVERY AFTER DISINFECTANT

7.2.1 Aseptically dispense required amount of disinfectant into sterile measuring cylinder (quantity
to be dispensed based on the concentration and volume to be prepared) and make up the
volume with sterile purified water to the required volume.
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

7.2.2 Distribute 10 ml of above prepared disinfectant into sterile test tubes and mark the tubes in
such a way mentioned below:
I test tube - 0 minutes contact time.
II test tube - 5 minutes contact time.
III test tube - 10 minutes contact time.
7.2.3 Inoculate 1 ml of culture suspension having more than 10000 cfu in to I tube (0 minutes)
7.2.4 Immediately shake the tube gently and carry out serial dilution in such a way to get less than
100 cfu count on a filter paper.
7.2.5 Filter the dilution (giving less than 100 cfu) through 0.45 µ membrane filter by applying
Vaccum .Rinse the filter paper twice with the 100 ml diluent.
7.2.6 Aseptically transfer the membrane on the surface of sterile Soyabean casein Digest Agar with
Tween 80.
7.2.7 In II test tube (5 min contact time), inoculate 1 ml of cultures suspension having more than
10000 cfu and allow it to stand for 5 minutes with intermediate shaking.
7.2.8 After 5 min carry out serial dilution in such a way to get less than 100 cfu count on a filter
paper.
7.2.9 Filter the dilution (giving less than 100 cfu) through 0.45 µ membrane filter by applying
Vaccum .Rinse the filter paper twice with the 100 ml diluent.
7.2.10 Aseptically transfer the membrane on the surface of sterile Soyabean Casein Digest Agar with
Tween 80.
7.2.11 Repeat the above procedure for 10 minutes
7.2.12 Repeat the above procedure for different culture suspension as required.
7.2.13 For bacterial cultures incubate the plates at 30-35° C for 3-5 days and fungal culture at 20-25
°C for 5-7 days.
7.2.14 Count the actual number of inoculated cfu in the diluent as above.
7.2.15 Negative Control: Carry out negative control by filtering the serial diluent through 0.45 µ
membrane filter paper and incubate the plate at 30-35°C for 3-5 days.

7.3 DISINFECANT VALIDATION SURFACE STUDY

7.3.1 CULTURE CONCENTRATION CONTROL
7.3.1.1 Take 50 -100 cm2 sterile surface area of above mentioned (Table-1) material surface
under LAF and mark 25 cm2 on it.
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

7.3.1.2 Aseptically inoculate evenly 1 ml (having more than 10000 cfu) with any one of the
culture suspension inside the marked 25 cm2 surface area of the material.
7.3.1.3 Under Laminar Air flow (LAF), dry the inoculated culture suspension material.
7.3.1.4 After completion of time period, inoculate the material in to sterile tube or beaker
containing sterile diluent and aseptically wipe the surface of material with sterile swab
gently to recover the organisms.
7.3.1.5 Shake the tube gently and carry out serial dilution in such a way to get less than 100
cfu count on a filter paper.
7.3.1.6 Filter the dilution (giving less than 100 cfu) through 0.45 µ membrane filter by
applying Vaccum .Rinse the filter paper twice with the 100 ml diluent.
7.3.1.7 Aseptically transfer the membrane on the surface of sterile Soyabean casein Digest
Agar with Tween 80.
7.3.1.8 For bacterial cultures incubate the plates at 30-35° C for 3-5 days and fungal culture at
20-25 °C for 5-7 days.
7.3.1.9 Count the actual number of inoculated cfu in the diluent as above.

7.4 MICROORGANISMS RECOVERY AFTER DISINFECTANT:

7.4.1 Take 50 -100 cm2 sterile surface area of above mentioned (Table-1) material surface under
LAF and mark 25 cm2 on it.
7.4.2 If material is autoclavable, sterilize the above material in steam sterilizer.
7.4.3 After sterilization aseptically transfer sterilized materials to Laminar Air flow (LAF).
7.4.4 If material is not autoclavable, carryout disinfection with approved disinfectant and rinse with
sterile purified water to remove disinfectant debris if any, before usage for validation studies.
7.4.5 Aseptically inoculate evenly 1 ml (having more than 10000 cfu) with any one of the culture
suspension inside the marked 25 cm2 surface area of the material.
7.4.6 Under Laminar Air flow (LAF), dry the inoculated culture suspension material.
7.4.7 After drying, apply disinfectant on the inoculated culture suspension and leave for time period
which is already determined in dilution study for particular disinfectant.
7.4.8 After completion of time period, inoculate the material in to sterile tube or beaker containing
sterile diluent and aseptically wipe the surface of material with sterile swab gently to recover
the organisms.
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

7.4.9 Shake the tube gently and carry out serial dilution in such a way to get less than 100 cfu count
on a filter paper.
7.4.10 Filter the dilution (giving less than 100 cfu) through 0.45 µ membrane filter by applying
Vaccum .Rinse the filter paper twice with the 100 ml diluent.
7.4.11 Aseptically transfer the membrane on the surface of sterile Soyabean casein Digest Agar with
Tween 80.
7.4.12 For bacterial cultures incubate the plates at 30-35° C for 3-5 days and fungal culture at 20-25
°C for 5-7 days.
7.4.13 Count the actual number of inoculated cfu in the diluent as above.
7.4.14 Repeat the above procedure for other remaining organisms and surfaces as mentioned in
Table-1based on the requirement.
7.4.15 Negative Control
7.4.15.1Take 50 -100 cm2 sterile surface area of above mentioned (Table-1) material surface
under LAF and mark 25 cm2 on it.
7.4.15.2Aseptically inoculate evenly 1 ml of sterile diluent inside the marked 25 cm2 surface
area of the material.
7.4.15.3Under Laminar Air flow (LAF), dry the inoculated diluent on the surface of the
material.
7.4.15.4After drying, aseptically wipe the surface of material with sterile swab and transfer it
into 10 ml diluent.
7.4.15.5Filter the diluent through 0.45 µ membrane filter by applying Vaccum .Rinse the
membrane filter twice with the 100 ml diluent.
7.4.15.6Aseptically transfer the membrane on the surface of sterile Soyabean casein Digest
Agar with Tween 80.
7.4.15.7Incubate the plate at 30-35°C for 3-5 days.
7.4.15.8Repeat the above procedure for other remaining surfaces as mentioned in Table- I
based on the requirement.
7.4.15.9 Acceptance Criteria: No growth should observed after incubation.

7.5 LOG REDUCTION CALCULATION:

Log Reduction = (Log N0 (Actual Number of inoculated organisms) – Log N (Number of organisms
recovered after disinfectant).
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

For example, if 12500 cfu of microorganisms are inoculated and recovered 05 cfu after disinfectant
application, the calculation shall be = log (12500) – log (5)
= 4.09 – 0.69
= 3.94 log reduction

8.0 VALIDATION TEST PROCEDURE FOR ESTABLISHMENT OF EFFECTIVENESS DURATION OF

DISINFECTANT SOLUTION AFTER APPLICATION IN CONTROLLED AND CLEAN AREA:
8.1 Use the environmental testing flexi plates (55 mm) to monitor the environmental microorganisms after
the application of disinfectant solution from the surfaces of floor of granulation area, Capsule Filling
room, Compression Room, Coating Room, Tablet Capsule inspection area, Solution preparation room,
Blister packing room and main corridor in the manufacturing area and floor and doors of MLT Room in
microbiology area.
8.2 Collect the sample from the above area after the application of disinfectant solution and then at different
location in the same area at a time interval of 02 hours, 04 hours, 06 hours and 08 hours.
8.3 Incubate the plates at 30-35°C for pre incubation to ensure that they are sterile before the test is carried
out.
8.4 On the day of monitoring, select the uncontaminated plates and mark the plates with location details.
8.5 Carefully open the plate and take the sample by pressing agar surface at specified location for a period of
5- 10 seconds for each plate.
8.6 Wipe the sampled area with 70 % filtered IPA solution immediately to remove the media traces from the
sampled area.
8.7 Incubate all the plates after testing at 20-25°C for a minimum of 72 hours. Further incubate all the plates
in inverted position for a period of 48 hours at 30-35°C.
8.8 ACCEPTANCE CRITERIA
8.8.1 VALIDATION TEST PROCEDURE FOR ESTABLISHMENT OF MINIMUM
EFFECTIVE CONCENTRATION OF DISINFECTANT
8.8.1.1 Minimum three log reduction of microorganisms should be obtained after disinfectant
application.
8.8.1.2 No growth should be observed in negative control.
8.8.2 VALIDATION TEST PROCEDURE FOR ESTABLISHMENT OF EFFECTIVENESS
DURATION OF DISINFECTANT SOLUTION AFTER APPLICATION IN
CONTROLLED AND CLEAN ROOM AREA
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

DISINFECTANT VALIDATION PROTOCOL AND REPORT

8.8.2.1 Total aerobic microbial count on the plate should not be more than 50 cfu/plate.

9.0 REFERENCES:
9.1 USP chapter <1072>

10.0 EVALUATION OF RESULTS

10.1 The decrease in the bacterial load to exposed disinfectant indicates that the disinfectant is capable of reducing the
contaminant when used in the area.

11.0 CONCLUSION
11.1 A summary report shall be prepared that contain discussion and conclusion with clearly state the successful
achievement of objective of validation studies.

12.0 ABBREVIATIONS:
SOP : Standard Operating Procedure
Ltd. : Limited
cfu : Colony Forming Unit
DV : Disinfectant Validation
W/V : Weight/ Volume
V/V : Volume/volume
QCD : Quality Control Department
LAF : Laminar Air Flow
IPA : Isopropyl alcohol
°C : Degree Centigrade
% : Percentage
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR VALIDATION OF DISINFECTANT SOLUTIONS – DILUTION METHOD

ANNEXURE I
Name of the Disinfectant Active Ingredient
Disinfectant Manufacturer Batch No./ Lot No. of Disinfectant
Disinfectant Concentration Used Membrane filter Lot No.
Media Used Media Lot No.
Diluent Used Media Lot No.
Incubator ID (32.5 ±2.5°C) Incubation Period
Incubator ID (22.5 ±2.5°C) Incubation Period
Date of Test Date of Report

OBSERVATION TABLE:
1. CULTURE CONCENTRATION CONTROL
(DILUENT + MICROBIAL CULTURE)
Stock Organism Recovery After Incubation
Stock
culture
Sr. Name of Culture Actual No. of Inoculated Cfu
concentrati
No Organisms Concentrati Plate 1 Plate 2 Average (Average cfu x Dilution Factor)
on dilution
on
times
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR VALIDATION OF DISINFECTANT SOLUTIONS – DILUTION METHOD

2.0 TEST OBSERVATION

MICROORGANISMS RECOVERY AFTER DISINFECTANT (DILUENT + DISINFECTANT + MICROBIAL CULTURE)
Organism Recovery Organism Recovery Actual No. of
Conta Organism Recovery
Stock After Incubation After Incubation Inoculated
After Incubation (1:10)
Sr. Name of Culture ct (1:100) (1:1000) Cfu
No Organisms Concentrati Plate Plate Averag Plate Plate Avera Plate Avera (Average cfu
on Time Plate 1 x Dilution
1 2 e 1 2 ge 2 ge
Factor)
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR VALIDATION OF DISINFECTANT SOLUTIONS – DILUTION METHOD

Organism Recovery Organism Recovery Actual No. of

Organism Recovery
Stock After Incubation After Incubation Inoculated
Conta After Incubation (1:10)
Sr. Name of Culture (1:100) (1:1000) Cfu
ct
No Organisms Concentrati (Average cfu x
Time Plate Plate Averag Plate Plate Avera Plate Avera
on Plate 1 Dilution
1 2 e 1 2 ge 2 ge
Factor)
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min

3.0 NEGATIVE CONTROL (DILUENT)

OBSERVATION:_______________________________________
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR VALIDATION OF DISINFECTANT SOLUTIONS – DILUTION METHOD

4. CALCULATION FOR LOG REDUCTION:

No. of cfu Recovered After
Sr. Actual No. of Inoculated Cfu
Contact Disinfectant
No Name of Organisms (Average cfu x Dilution Factor) Log Reduction = Log N0 – Log N
Time (Average cfu x Dilution Factor)
(N0)
(N)
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR VALIDATION OF DISINFECTANT SOLUTIONS – DILUTION METHOD

No. of cfu Recovered After

Actual No. of Inoculated Cfu
S. Name of Contact Disinfectant
(Average cfu x Dilution Factor) Log Reduction = Log N0 – Log N
No Organisms Time (Average cfu x Dilution Factor)
(N0)
(N)
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min

ANALYSED BY CHECKED BY APPROVED BY

Name:
Designation:
Signature :
Date:
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR VALIDATION OF DISINFECTANT SOLUTIONS – SURFACE METHOD

ANNEXURE II
Name of the Disinfectant Active Ingredient
Disinfectant Manufacturer Batch No./ Lot No. of Disinfectant
Disinfectant Concentration Used Membrane filter Lot No.
Media Used Media Lot No.
Diluent Used Media Lot No.
Incubator ID (32.5 ±2.5°C) Incubation Period
Incubator ID (22.5 ±2.5°C) Incubation Period
Date of Test Date of Report
Surface Used

OBSERVATION TABLE:
1. CULTURE CONCENTRATION CONTROL
(DILUENT + MICROBIAL CULTURE)

Stock Organism Recovery After Incubation

Stock
culture
Sr. Name of Culture Actual No. of Inoculated Cfu
concentrati
No Organisms Concentrati Plate 1 Plate 2 Average (Average cfu x Dilution Factor)
on dilution
on
times
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR VALIDATION OF DISINFECTANT SOLUTIONS – SURFACE METHOD

2. TEST OBSERVATION
MICROORGANISMS RECOVERY AFTER DISINFECTANT (DILUENT + DISINFECTANT + MICROBIAL CULTURE)
Organism Recovery Organism Recovery Actual No. of
Organism Recovery
Stock After Incubation After Incubation Inoculated
Conta After Incubation (1:10)
S. Name of Culture (1:100) (1:1000) Cfu
ct
No Organisms Concentrati Plate Plate Plate Plate Plate (Average cfu
Time Avg. Avg. Plate 1 Avg.
on x Dilution
1 2 1 2 2
Factor)
0 min
1 5 min
10 min
0 min
2 5 min
10 min
0 min
3 5 min
10 min
0 min
4 5 min
10 min
0 min
5 5 min
10 min
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR VALIDATION OF DISINFECTANT SOLUTIONS – SURFACE METHOD

Organism Recovery Organism Recovery Actual No. of

Organism Recovery
Stock After Incubation After Incubation Inoculated
Conta After Incubation (1:10)
Sr. Name of Culture (1:100) (1:1000) Cfu
ct
No Organisms Concentrati (Average cfu
Time Plate Plate Plate Plate Plate
on Avg. Avg. Plate 1 Avg. x Dilution
1 2 1 2 2
Factor)
0 min
6 5 min
10 min
0 min
7 5 min
10 min
0 min
8 5 min
10 min
0 min
9 5 min
10 min
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR VALIDATION OF DISINFECTANT SOLUTIONS – SURFACE METHOD

3. NEGATIVE CONTROL (DILUENT)

OBSERVATION:

S.No. Surface Details Observation

 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR VALIDATION OF DISINFECTANT SOLUTIONS – SURFACE METHOD

4.0 CALCULATION FOR LOG REDUCTION

No. of cfu Recovered After

Actual No. of Inoculated Cfu
Sr. Contact Disinfectant
Name of Organisms (Average cfu x Dilution Factor) Log Reduction = Log N0 – Log N
No Time (Average cfu x Dilution Factor)
(N0)
(N)
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR VALIDATION OF DISINFECTANT SOLUTIONS – SURFACE METHOD

No. of cfu Recovered After

Actual No. of Inoculated Cfu
Sr. Contact Disinfectant
Name of Organisms (Average cfu x Dilution Factor) Log Reduction = Log N0 – Log N
No Time (Average cfu x Dilution Factor)
(N0)
(N)
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min
0 min
5 min
10 min

ANALYSED BY CHECKED BY APPROVED BY

Name:
Designation:
Signature :
Date:
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR DISINFECTANT VALIDATION

ANNEXURE III
Name of the Disinfectant
Active Ingredient
Batch No./ Lot No. of Disinfectant Disinfectant Concentration Used
Name of Area Sampled By
Date of Sampling Time of Sampling
Name of Media Soyabean Casein digest Agar Media Lot No
Incubation Temperature 22.5 ± 2.5°C Incubation Period
Incubation Temperature 32.5 ± 2.5°C Incubation Period
Incubator ID (22.5 ± 2.5°C) Incubator ID (32.5 ± 2.5°C)
Date of Report

Sr.
Name of Location Sampled Area Total Aerobic Microbial Count (cfu/plate)
No
 PHARMA DEVILS
MICROBIOLOGY DEPARTMENT

ANALYTICAL WORKSHEET FOR DISINFECTANT VALIDATION

S.No Name of Location Sampled Area Total Aerobic Microbial Count (cfu/plate)

Negative Control
Total Aerobic Microbial Count Limit
TAMC Limit NMT 50 cfu/Plate

Remark:_________________________________________________________________________________________

ANALYSED BY CHECKED BY APPROVED BY

Name:
Designation:
Signature :
Date: