IL-Cleaning Validation

CLEANING OF SURFACE AREA VALIDATION REPORT
INTERTECH LABS
CLEANING OF
SURFACE AREA VALIDATION
PROTOCOL
INTERTECH LABS
CLEANING OF
PROTOCOL
CLEANING (SURFACE) VALIDATION

INTERTECH LABS
CLEANING OF
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REPORT
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INTERTECH LABS
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TABLE OF CONTENTS
S. No. TITLE Page No.
1.0 Report approval 03

2.0 Objective 04
3.0 Scope 04
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4.0 Abbreviation 04
5.0 Responsibility 05
6.0 Training verification 05-06
7.0 Materials and methods 06-10
8.0 Procedure 11-13
9.0 Deficiency and corrective action report 13
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10.0 Summary and conclusion 14
11.0 Annexure 14
12.0 Reference documents 14
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1.0 REPORT APPROVAL:
1.1. Signing of this approval page of protocol indicates agreement with approach described in
this document. If any modifications in this approach and document become necessary, an
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addendum shall be prepared, checked and approved. This protocol cannot be executed until
approved
2.0. OBJECTIVE:
Preparation of report for the executed for cleaning validation of surfaces disinfection with dettol,savlon
and Virosil.
3.0. SCOPE:
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The scope include surface Cleaning and sanitization validation of floor,walls,roof with 3 sanitizing
agents used in rotation.
4.0. ABBREVIATION:
ATCC American Type Culture collection L Liters
LAF Laminar Air Flow NA Not Applicable
CAPA Corrective and Preventive Action mm Millimeter
cGMP Current Good Manufacturing mg Milligram
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practices
CFU Colony Forming Units NCIM National collection of Industrial Microorganism
NMT Not More than NA Not Applicable
°C Degree Celsius NLT Not Less than
IPA Isopropyl alcohol NMT Not More than
GTP General test procedure SCDA Soyabean Casein Digest Agar
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5.0 RESPONSIBILITY OF VALIDATION TEAM:

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S.No Responsibility Resposibilty discharge

1. For preparation of the protocol.
The responsibilities
1 2. Monitoring as per protocol. assigned in the
3. Assist in case of any failures. protocol or discharged
The responsibilities
2 1. Execution of the protocol. assigned in the
protocol or discharged
1. For review and approval of the

protocol. The responsibilities
2. Monitoring of the cleaning validation assigned in the
3 protocol or discharged
as per protocol.
3. Approval of the protocol.
4. Notes Deviations / Correctness
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6.0. TRAINING RECORD OF SOP AND PROTOCOL:

Responsible people were trained including ,microbiologist on the cleaning validation protocol.
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6.1. Execution/ Working Team:
The execution team have discharged the responsibility and carried out the cleaning validation study as
prescribed in the protocol without any deviation ,changes.
Execution team signatures

S.No. Name Signature Remarks
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1. 1
2. 2
3. 3
4. 4
5. 5
6.2. The following reference SOP/STP were referred vial execution media fill Attach the copies of
SOP/STP
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S.No. SOPs/ STPs for referral Procedures

1. Selection and use of cleaning agents and disinfectants and their 4.2.1
concentrations at sterile area and non sterile areas.
2. Cleaning procedure for compounding room 4.2.2
3. Procedure for swab test (equipment surfaces, sterile garments, false 4.2.3
ceiling and personnel hygiene).
4. Media growth promotion, inhibition and sterility check 4.2.4
5. Preparation and preservation of 10-100 cfu’s/0.1 ml sub culture 4.2.5
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suspension
6. Isolation and identification of isolates 4.2.6
Operation and calibration of bacteriological incubator 4.2.7
7.
(Newtronics)
7.0. MATERIALS AND METHODS USED:
The disinfectants media subculture was used as materials, Microbiology test methods- MB/024,
MB/018, MB/011 and MB/015. were used. The –doc no 058 were used.
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7.1. Cultures:
7.1.1. The following Microbial subcultures are used during the validation:
Table-1
S. No. Microorganism NCIM / ATCC Number Incubation conditions
Bacterial Cultures
01 Staphylococcus aureus NCIM 5345 / ATCC 6538 30ºC - 35 ºC / 3Days
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02 Pseudomonas aeruginosa NCIM 2200 / ATCC 9027 30ºC - 35 ºC / 3Days
03 Escherichia coli NCIM 2065 / ATCC 8739 30ºC - 35 ºC / 3Days
04 Bacillus subtilis NCIM 2063 / ATCC 6633 30ºC - 35 ºC / 3Days
Fungal Cultures
05 Candida albicans NCIM 3471 / ATCC 10231 20ºC - 25 ºC / 5Days
7.2. Media:
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7.2.1. Sterile peptone and Soyabean Casin Digest Agar were used for inoculation and incubation.
7.3. Disinfectants concentrations used for surface disinfection by rotation.

Table-2
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S. Name of the Concentrati Mode of Mode of Diluents

Disinfectant on Chemical Composition
No. Application Action Sterile
1 Dettol 2.5 % v/v Chlorxylenol and Terpineol water
Surface Antiseptic
are mainly constituents of
Disinfectant Liquid
Dettol.
Rational As per the label of the bottle it is advised to dilute in water hence we are using WFI for
dilution
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S. Name of the Concentrati Mode of Mode of

No. Application Action
2 Savlon 2.5 % v/v Aqua, Sodium Laureth Disinfectant
Surface
Sulfate, Cocamide MEA and surface
Disinfectant
Sodium Chloride cleaner
Rational As per the label of the bottle it is advised to dilute in water hence we are using WFI for Diluents
Sterile
dilution water
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S. Name of the Concentrati Mode of Mode of

No. Application Action
3 Virosil 10.0 % v/v Complex formulation of Diluents
Surface
Stabilized Hydrogen Peroxide Antiseptic Sterile
Disinfect water
11% w/v with 0.01% w/v Diluted Disinfectant.
ant
Silver Nitrate Solution water Q.S.
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7.3.1. The disinfectants are evaluated in this study are in table 2

7.3.2. The above disinfectants are selected based on the following factors.
7.3.3. Based on broad spectrum of anti microbial properties, miscibility in water,Scentific literature,
published by manufacturers of 3 disinfectants which are attached to the report .
7.3.4. The following Contact times of above disinfectants were allowed during cleaning
5 min 10min 15 min.
7.3.5. PPE were used all the persons involved in cleaning and disinfectant usage..
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7.3.6. Note: All above disinfectants are as per above concentrations (as per manufacturer label indications
the above disinfectant concentrations are concluded)
7.4. Following Equipments / instruments are used as below:

7.4.1.Bio-safety cabinet / Laminar Air Flow
7.4.2.Incubators (20-25ºC & 30-35ºC)
7.4.3.Colony counter
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7.4.4.Sterile filtration membranes (0.45µ) and filtration unit.

7.4.5.Micropipettes & sterile tips
7.4.6.Sterile forceps
7.4.7.Sterile containers
7.5.Following Location:
7.5.1. Disinfectant for validation as per given table as above.

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7.5.2. Disinfectant validation study is carried out on below mentioned surfaces
Table-3
S.No. Location
1.0 Floor cleaning in all 17 clean rooms as per doc 028
2.0 Ceiling and Walls of clean rooms as per doc 028
7.6. Cleaning Procedure Followed as per SOP –IPR/058 which is as below:

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7.6.1. Production area consists of Process area, general area and packing area.
7.6.2. Type of Area Cleaning: Area cleaning is defined on the basis of requirement of area Cleanliness.
7.6.3. Dry Cleaning: Hand pick unnecessary items (e.g. damaged polybags, gaskets, metal pieces, paper
etc)if found. Clean the floor by mechanical sweeper or by nylon broom. Clean exterior of
material container, drums manually by wiping with cotton cloth. Clean Overhead ducts, ceiling
and walls by long handled brushes/ mop.
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7.6.4. Wet Cleaning: Clean the area with wet cotton wiper / mop / mechanical sweeper moistened with
disinfectant. Use any disinfectants out of the listed four. Rotate disinfectant after every one in day.
3 types of cleaning defined based on criticalities

S.No. Type of cleaning Method of cleaning Type of Area to be cleaned
1. I Cleaning and wet All the change rooms material entry men
mopping with Dettol / entry, Packing Corridor, Packing rooms,
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Savlon/ Virosil. compounding room, Blending room, Filling

rooms, Container washing rooms etc----
That is clean room-B,C,D Areas and
Hygiene area .
2. II Wet mopping with Dettol Doors and windows, corridor’s, WFI.
/ Savlon/ Virosil
3. III Brooming and Dry Control panel room etc
mopping
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7.7. Sampling for Validation:
7.7.1. following procedure implemented and done :

A. Type of swab.
B. Absorbent cotton swab of 5 cm. × 5 cm. (25 cm2) shall be used.
C. Selection of worst case.(Angular corners and other corners, Coving edges, etc).
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7.7.2. Sampling diagram:

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7.8. Cleaning Visual Inspection verification:

7.8.1. After the cleaning of Area and location, visual inspection was performed prior to other sampling., it
was visually clean. Hence swabs were taken.
8.0. PROCEDURE are Followed as below:

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8.1. Disinfectant cleaning Validation by swab Method by sterile swabs

8.2. Preparation of Inoculum determination
8.3. Selection of Disinfectant Concentration
8.4. Neutralizer Efficacy Study
8.5. Negative Product Control
8.6. Positive Control
8.7. Determining the Efficacy of the Disinfectant By Tube Dilution Method
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8.8. Negative Control

8.9. Preparation of Spore Suspension
9.0. PROCEDURE:
9.1. DISINFECTANT CLEANING VALIDATION BY SWAB METHOD BY STERILE SWABS:
9.1.1. Swab method followed is as below:

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This method is used for Swab samples taken as per locations given in cleaning validation protocol. Swab
stick was used for sampling. Defined volume of solvent in which active ingredient is soluble, taken in test
tube. Deep the swab stick into it and wipe on 5 x 5 cm surface area of equipment in the manner given
below. Deep the swab sticks into test tube and cover the test tube. Repeat the procedure for the other
locations.
9.2. INOCULUM was prepared as below :

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9.2.1. Followed the preparation of inoculum Preparation as per SOP: MB/014 “Receipt, storage, revival
lot preparation, subsequent sub culturing of master cultures and inoculum preparation”.
9.3. SELECTION OF DISINFECTANT CONCENTRATION WAS DONE AS PER

MANUFACTURER INSTRUCTIONS AND LITERATURE
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9.3.1.Manufacturer's recommended concentration was checked for use by referring the literatures
supplied by the manufacturer.
9.3.2. Prepared the disinfectants at different concentrations as per the below tables and record the details
in Annexure-I (Record for preparation of disinfectants).
Table-4
Disinfectant concentration and contact time details
S. No. Name of the Required concentration of Disinfectant
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Disinfectant Day Actual

Low concentration High concentration
concentration
1 Dettol 2.5% v/v Day-1 2.5% 1.5% 3.5%
2 Savlon 2.5%v/v Day-2 2.5% 1.5% 3.5%
3 Virosil 10.0%v/v Day-3 10.0% 8.0% 12.0%
Contact time 5 min, 10 min and 15 min.( Each concentration)
Table-5
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Disinfectant For 100 ml preparation

S. No Name of the Disinfectant Actual
Quantity of For 100 ml
concentration
disinfectant preparation
1 Dettol 2.5% v/v 03 ml 97 ml
2.5 %
2 Savlon 2.5%v/v 2.5 ml 97.5 ml
2.5 %
3 Virosil 10.0%v/v 10.0 % 10 ml 90 ml
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S. No Low
Name of the Disinfectant Quantity of For 100 ml
concentration
1 Dettol 2.5 % v/v 02 ml 98 ml
1.5%
2 Savlon 2.5 %v/v 1.5 ml 98.5 ml
1.5%
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3 Virosil 10.0 %v/v 8.0% 8 ml 92 ml

S. No Name of the Disinfectant High
Quantity of For 100 ml
concentration
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1 Dettol 2.5 % v/v 04 ml 96 ml

3.5%
2 Savlon 2.5 %v/v 3.5 ml 96.5 ml
3.5%
3 Virosil 10.0 %v/v 12.0% 12 ml 88 ml
9.3.4. Original disinfectant diluted to the predetermined concentration level in a sterilized container by
using WFI( water for injection) filtered thru 0.22um filter
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9.3.5.Filtered disinfectant solution collected in a sterilized container and label the container which
represent the Name of disinfectant, Concentration, Date of preparation, use before and prepared by
(Sign/Date).
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9.5. NEGATIVE CONTROL:

9.5.1. Pre-wet the membrane with 1×50 mL quantity of sterilized 0.1% of peptone water.
9.5.2. Then membrane shall be rinsed with 3 x 100 mL of 0.1% peptone water.
9.5.3. Membrane filter shall be transferred aseptically on to the surface of pre- incubated SCDA / SDA
plate.
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9.5.4. SCDA plate shall be incubated at,30-35 ºC for 3 to 5 days and SDA plate shall be
Incubated at 20-25 ºC for 5 to 7 days.
9.5.5The test details and results shall be recorded in Annexure
10.0. DEFICIENCY AND CORRECTIVE ACTION REPORT

No Deviations, No Changes No Deficiencies noted during the execution of cleaning validation
study in aseptic injection clean rooms.
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11.0. SUMMARY AND CONCLUSION

1.Validation study was carried out as per the protocol without any Deviations and Changes
2.Disinfectants were rotated one after another for 3 days.
3.The cultures at the concentrations were 5 ml/100 were spread in the surfaces and swabbed
after disinfectant and cleaning as per SOP-IPR/058.
4. Sterile swabs were used to swab the surface after cleaning.
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5.Swabs were extracted in to relevant solutions inoculated and incubated and results are
tabulated
Hence cleaning validation of surfaces was done as per protocol prescription and concluded as
validated. Since concenratarions dettol &savalon are 2.5% and virosil 10% is desirable effect
hence desirable doc 058 can be followed for cleaning
12.0 ANNEXURES:
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12.1. Annexure-I: Report for preparation of disinfectants

12.2. Annexure-II: Test results of all surfaces worst case criteria + one random.
13.0 REFERENCE:
13.1. USP1072: “Disinfectants and antiseptics”

13.2 Cleaning of Surface doc no058
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13.6. Cleaning procedure doc 042

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CLEANING OF SURFACE AREA VALIDATION REPORT

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

CLEANING (SURFACE) VALIDATION

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

S. No. TITLE Page No.

1.0 Report approval 03

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

10.0 Summary and conclusion 14

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

1.0 REPORT APPROVAL:

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

5.0 RESPONSIBILITY OF VALIDATION TEAM:

SURFACE AREA VALIDATION

S.No Responsibility Resposibilty discharge

1. For review and approval of the

SURFACE AREA VALIDATION

6.0. TRAINING RECORD OF SOP AND PROTOCOL:

SURFACE AREA VALIDATION

6.1. Execution/ Working Team:

Execution team signatures

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

S.No. SOPs/ STPs for referral Procedures

SURFACE AREA VALIDATION

7.0. MATERIALS AND METHODS USED:

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION

02 Pseudomonas aeruginosa NCIM 2200 / ATCC 9027 30ºC - 35 ºC / 3Days

03 Escherichia coli NCIM 2065 / ATCC 8739 30ºC - 35 ºC / 3Days

04 Bacillus subtilis NCIM 2063 / ATCC 6633 30ºC - 35 ºC / 3Days

SURFACE AREA VALIDATION

7.3. Disinfectants concentrations used for surface disinfection by rotation.

SURFACE AREA VALIDATION

S. Name of the Concentrati Mode of Mode of Diluents

SURFACE AREA VALIDATION

S. Name of the Concentrati Mode of Mode of

SURFACE AREA VALIDATION

S. Name of the Concentrati Mode of Mode of

SURFACE AREA VALIDATION

7.3.1. The disinfectants are evaluated in this study are in table 2

SURFACE AREA VALIDATION

7.4. Following Equipments / instruments are used as below:

SURFACE AREA VALIDATION

7.4.4.Sterile filtration membranes (0.45µ) and filtration unit.

7.5.1. Disinfectant for validation as per given table as above.

SURFACE AREA VALIDATION

7.5.2. Disinfectant validation study is carried out on below mentioned surfaces

7.6. Cleaning Procedure Followed as per SOP –IPR/058 which is as below:

SURFACE AREA VALIDATION

SURFACE AREA VALIDATION