International journal of basic and applied research

www.pragatipublication.com
ISSN 2249-3352 (P) 2278-0505 (E)
Cosmos Impact Factor-5.86

Evaluation study for determination of quantities of disinfectant solution used

for fogging of clean rooms with respect to area volume

Sachin D. Joshi1, Dr. Imran Memon, Dr. Tahur Shaikh, Idris Khan and Surjeet Samanta
IMAGO & GETTER, Mumbai

Abstract: The evaluation study was aimed to determine the total quantity of disinfectant solution to be
used with respect to the size of area for fogging. Now a days fogging is used to control the microbial
contamination in controlled area. Fogging is a process where chemical disinfectants are applied to
production areas, clean rooms, sterile areas, operation theatre etc. in Pharma industries, Institution,
Healthcare, etc. The purpose is to create and disperse a disinfectant aerosol to reduce the numbers of
airborne micro-organisms, to prevent transmission of potential contaminants and also to apply
disinfectant to surfaces that may be difficult to reach [3]. Critical factors to address while fogging is
the quantity of disinfectant solution to be used in different area sizes [6]. Therefore, calculation for
volume of diluted disinfectant solution to be used during fogging for different area sizes has been
established by a fast and simple method is presented and discussed.

Keywords: Fogging, Pharma industries, volume of disinfectant, area size, clean rooms, aerosol and
contaminants.

Introduction
Cleaning and disinfection practices are an essential part of contamination control in the
pharmaceutical industry and in healthcare [4]. During manufacture, products can be exposed to
microbiological cross-contamination from surfaces and the air, which may give rise to product
spoilage and safety issues. The traditional approach to controlling such contamination has been to
implement cleaning and disinfection regimes. This targeted approach may be sufficient to maintain
day-to-day control of contamination, but does not necessarily eliminate all environmental micro-
organisms and in some instances, they can persist in factories for several years. To keep the
controlled area from being contaminated in pharmaceutical industry and healthcare, two processes
namely fumigation and fogging are used [2].
Fumigation is banned by different regulatory agencies in many pharmaceuticals and healthcare
because of its negative effects such as it involves formaldehyde solution, which is carcinogenic in
nature, also it causes the user irritation of eyes and nose leading to dizzy head and nausea, fumigation
also requires lot of cleaning up after the process is implemented, it generally requires de-fumigation
or air handling unit (AHU) to be continuously run for few hours in order to remove the residue from air,
etc. While fumigation has so many drawbacks, fogging is completely safe and is used more than
fumigation [1] [2].
Fogging is a technique which requires a fogger machine, which effectively fills the space with the
fogging solution (disinfectant solution). Fogging is done to sterilize the air of controlled area. The
fogger machine simply sprays the disinfectant solution in the form of aerosol or mist in the controlled
507 Received: 8 December Revised: 17 December Accepted: 24 December
Index in Cosmos
January 2019 Volume 9 Number 1
UGC Approved Journal
 International journal of basic and applied research
www.pragatipublication.com
ISSN 2249-3352 (P) 2278-0505 (E)
Cosmos Impact Factor-5.86

area. The small particles of disinfectant solution suspend in the air for long time and kill all the air
borne microorganisms which make it very effective way to control the contamination. Both the
pharmaceutical and clinical sectors have been targeted by a range of whole room type
decontamination systems, but their practical operation and benefit in the industry is relatively
unknown [3]. During practical operation of fogging, the problem arises that how much disinfectant
solution to be used for a particular size or room area. Thus this increasing problem of disinfectant
during fogging across the Pharma manufacturing industry, institution, healthcare etc. made necessary
for this study and is critically discussed as some microorganisms may build up resistance phenomena
[2]. This study provides a ready guidance for disinfection of controlled areas.

Materials and methods

Disinfectant:
ENVISHIELD disinfectant product containing Hydrogen peroxide and Silver ions solution, for Surface
and Environment Disinfection (from Imago & Getter, Mumbai)
Preparation of Standard Dilution:

Table 1: ENVISHIELD disinfectant was diluted as 20% in sterile distilled water.

Envishield (to prepare20% Volume of WFI in ml Total Volume of Disinfectant
solution) Fogging Solution in ml
200 800 1000
400 1600 2000
600 2400 3000
800 3200 4000
1000 4000 5000
Apparatus:
A SKANFOG vapor jet ULV (Ultra Low Volume) fogger machine (Imago & Getter, Mumbai) for
aerial disinfection, consisting of a 5 liters high grade chemical solution tank with material of
construction as HDPE or SS 304 which is easy to clean, detachable, non-corrosive and non-reactive.
Equipped with superior quality double stage, tapered fan, vacuum motors, non-clogging vertex
design of nozzle and suitable design with double layers for primary & secondary filtration to prevent
entry of dust / dirt & moisture from the air. It has uniform spreading and excellent control on droplet
size which generates aerosols in the range of 0.3 to 1 micron and discharge capacity of 50ml/min.
The different size or room area is selected on the basis of Area volume i.e.
Length (ft) x Width (ft) x height (ft) = Area volume in cubic feet.
Method:
Based on different area volume and application rate as given in Table 2 was selected for
fogging process. Application rate is based on the fogger flow rate. Total volume of disinfectant
solution of 20% dilution was prepared in sterile water as per in Table 1. The total volume of
disinfectant solution of 20% was calculated as per followin given in Table 2 [6].
Total quantity of disinfectant solution (ml) = volume of area x application rate (ml/1000 cubic meter)

508 Received: 8 December Revised: 17 December Accepted: 24 December

Index in Cosmos
January 2019 Volume 9 Number 1
UGC Approved Journal
 International journal of basic and applied research
www.pragatipublication.com
ISSN 2249-3352 (P) 2278-0505 (E)
Cosmos Impact Factor-5.86

Table 2. Calculation of Total volume of disinfectant solution of 20% dilution

Volume of area Application rate Total quantity of
(cubic feet) (ml/1000 cubic meter) disinfectant solution (ml)

984 400 393.6 (approx. 500ml)

800 787.2 (approx. 1000ml)

1865 400 746 (approx. 1000ml)

800 1492 (approx. 1500ml)

2700 400 1080 (approx. 1000ml)

800 2160 (approx. 2000ml)

All the door and windows were kept closed; AHUs were shut off making sure that all the
surfaces in area are exposed to disinfectant. Total volume of dilution concentration of disinfectant in
water was prepared as given in Table 2, for the selected volume of area [5]. Diluted disinfectant
solution was added into the fogger machine tank. The fogger machine was then placed at one corner
of the area. The number of fogger machine required may depend on size of the area to be fogged.
Flow rate is preset in the fogger machine and fogging time is calculated as [6]
Fogging time (min.) = Total quantity of Disinfectant solution / flow rate
Flow rate taken is 100ml/min.
The fogging process was done as per the time calculated for each area size. After completion
of the activity, continuous circulation of AHU (Air Handling Unit) was done for 6 hours. Fogging
process is validated to verify the effectiveness of the fogging and ensure that whole room disinfection
has been done properly. It was validated using Chemical indicator (Hydrogen peroxide detection)
and Biological indicator (Geobacillus stearothermophillus ATCC 7953 having 106 populations) strips.
Positive control and negative control was also kept for both the indicator strips. Once fogging
procedure is validated in one area, the same procedure was applicable to other areas [4] [5].
Method validation:
The method was validated using Chemical indicator (Hydrogen peroxide detection) and Biological
indicator (Geobacillus stearothermophillus ATCC 7953 having 106 populations) strips which were
placed in the area at different sites before fogging. A room area layout was prepared in which the
location of placement of Chemical indicator and Biological indicator strips were properly marked and
indicated. After performing the fogging according to designated time, the Chemical Indicator strips
was removed & checked for color change with control strip. Also the exposed Biological indicators
were transferred to microbiology laboratory in closed container. Aseptically the Biological indicators
strips were removed under Bio safety cabinet by using sterile forceps and transferred to test tube
containing 10 ml sterile Soyabean casein digest broth medium. The test tubes were incubated at 55 to
60ºC for 48-72 hrs. After incubation growth in test tubes was observed and recorded [4] [7].

509 Received: 8 December Revised: 17 December Accepted: 24 December

Index in Cosmos
January 2019 Volume 9 Number 1
UGC Approved Journal
 International journal of basic and applied research
www.pragatipublication.com
ISSN 2249-3352 (P) 2278-0505 (E)
Cosmos Impact Factor-5.86

Results and discussion

Each Chemical indicator strips were observed for colour change (e.g. from Magenta to yellow
colour) to interpret that if No Colour change (NC) is observed then Disinfection is not achieved and if
Colour change (C) is observed it means Disinfection is achieved.
For Biological indicator strips growth in test tubes were observed as if broth turns turbid, it
indicates that Disinfection is not achieved and if broth remains clear it means Disinfection is achieved.
The results for the area fogged for disinfection is summarized as in Table 3.

Table 3: Results of Disinfection validation

Volume of Application Total Fogging Chemical Biological
area rate quantity of time indicator indicator
(cubic feet) (ml/1000 cubic disinfectant (minutes)
meter) solution
984 400 500 ml 5 min. NC +++

800 1000 ml 10 min. C -

1865 400 1000 ml 10 min. NC ++

800 1500 ml 15 min. C -

2700 400 1000 ml 10 min. NC ++

800 2000 ml 20 min. C -

Where,
For Chemical indicator For Biological indicator
NC - No Colour change + = Turbid solution
C – Colour change - = Clear solution

Discussion
The results obtained in Table 3 showed that when total quantity of disinfectant solution
calculated with respect to 400ml / 1000 m3 showed No colour change in chemical indicator strips and
growth (turbidity) were observed in Biological indicator test tubes which indicates poor disinfection
of the area whereas the total quantity of disinfectant solution calculated with respect to 800ml / 1000
m3 showed Colour change in chemical indicator strips and no growth (Clear) were observed in
Biological indicator test tubes which indicates excellent disinfection of the area. It can be summarized
as given in Table 4.

510 Received: 8 December Revised: 17 December Accepted: 24 December

Index in Cosmos
January 2019 Volume 9 Number 1
UGC Approved Journal
 International journal of basic and applied research
www.pragatipublication.com
ISSN 2249-3352 (P) 2278-0505 (E)
Cosmos Impact Factor-5.86

Table 4. Quantity of Disinfectant solution v/s Area (cu.ft.) for fogging

Area Coverage Total quantity of Fogging time
(cu.ft.) disinfectant solution (minutes)

1000 1.0 Liters 10 min.

2000 1.5 Liters 15 min.

3000 2.0 Liters 20 min.

4000 2.5 Liters 25 min.

5000 3.0 Liters 30 min.

6000 3.5 Liters 35 min.

7000 4.0 Liters 40 min.

8000 4.5 Liters 45 min.

9000 5.0 Liters 50 min.

10,000 5.0 Liters 60 min.

Conclusion
Properly designed, appropriately qualified and consistently executed disinfection
procedures are critical to the production of safe and effective products. We presented a simple
method to calculate the total quantity of disinfectant solution for fogging, the application rate of the
Envishield disinfectant to be used should be 800ml / 1000 m 3 for different room volume at flow rate of
100ml/min. preset in the fogger machine. Customers could utilize the dry fog disinfection process on
a regular weekly basis without incurring large labor costs or tying up the area for long time periods.
Thus it gives the users such as Pharma industries, Healthcare, Institution etc. a ready reference to
calculate the total quantity of disinfectant needed and time interval required for fogging of room or
area to be disinfected. Careful review of the results in this study will help end users to monitor
potential deficiencies in their cleaning and disinfection program. As a result of this study it will allow
the customers to investigate and re-establish environmental control ultimately ensuring a safer
product for the end user.
References
[1] D. Vincent, (2002), ‘Validating, Establishing and Maintaining A Routine Environmental Monitoring
Program for Cleanroom Environments: Part 1’, Journal of Validation Technology, Vol. 8, No.4.

[2] E. Bessems, (1998), ‘The effect of practical conditions on the efficacy of disinfectants’, International
Biodeterioration and Biodegradation (41), pp:177-183.

511 Received: 8 December Revised: 17 December Accepted: 24 December

Index in Cosmos
January 2019 Volume 9 Number 1
UGC Approved Journal
 International journal of basic and applied research
www.pragatipublication.com
ISSN 2249-3352 (P) 2278-0505 (E)
Cosmos Impact Factor-5.86

[3] P. Vina, S. Rubio and T. Sandle (2011): ‘Selection and Validation of Disinfectants’, in New Delhi:
Business Horizons, pp:219-236.

[4] R. Klinkerberg, B. Streel and A. Ceccato, (2003), Development and validation of a liquid
chromatography method. J. Pharm. Biomed. Anal. (32), pp:345–352.

[5] R. D. Biunayki, et.al (2014), Strategies for the assessment of Disinfection and Cleaning on
Biopharmaceutical Cleanroom. Advances in Biomedical Engineering Research (ABER) Volume 2,
pp:18-27.

[6] T. Sandle (2012). ‘Application of Disinfectants and Detergents in the Pharmaceutical Sector’. In
Sandle, T. (2012). The CDC Handbook: A Guide to Cleaning and Disinfecting Cleanrooms, Grosvenor
House Publishing: Surrey, UK, pp168-197.

[7] US Environmental Protection Agency Office of Pesticide Programs, (2015), Protocol for Room
Sterilization by Fogger Application. pp:1-8

512 Received: 8 December Revised: 17 December Accepted: 24 December

Index in Cosmos
January 2019 Volume 9 Number 1
UGC Approved Journal