How to prove 6 % Hydrogen peroxide (H2O2) Suitable for Fogging activity

·         Calculate the required quantity of 6 % Hydrogen peroxide (H₂O₂) to be prepare.

·         And add the required quantity of water(purified water).

·         For example 100 ml of 6 % Hydrogen peroxide (H₂O₂) shall be prepared by using  30 % concentrated Hydrogen peroxide (H₂O₂) as per below.

Volume of to be fogged = length x width x height

Calculation total qty of liquid

=     liquid (ml) = volume X application rate (800 ml/ 1000 cubic meter(M³))

=       1464 X 800/1000 = 1171 ( 1200 ml)

Calculate Fogging Time:

=       total liquid / flow rate

=     1200/ 70 = 17.14 minutes (20 minutes)

Volume of Hydrogen peroxide (H₂O₂) = 1200 / 30 %    X 6 %  = 240 ml

Volume of water = 1200-240 = 960 ml

·         Inform the Engineering person to switch off the AHU through inter office communication.

·         Hang the SS carrier biological indicators / Self contained biological indicators with at the top critical corners of the room ( base on the risk assessment).

·         Before stat the fogging hang the biological indicators.

Sampling locations:

Sample No	Room Name	Location
1	Change Room-1	Top corner of the room near the dress cabinet.
2	Change Room-2	Top corner of the room near the Garment Cubicle.
3	Change Room-3	Top corner of the room near controlled area corridor.
4	Controlled area Corridor	Top corner of the room near the dynamic passbox.
5		Top corner of the room near the cool zone door.
6	Cool zone	Top corner of the room near the dynamic passbox.
7		Top corner of the room near the View glass.
8	Culture Handling Room	Left front top corner of the room.
9		Left rear top corner of the room.
10		Right rear top corner of the room.
11		Right front top corner of the room.
12		In side the Bio-safety cabinet left rear top corner
13		In side the Bio-safety cabinet right rear top corner
14	Testing area	Left front top corner of the room.
15		Left rear top corner of the room.
16		Right rear top corner of the room.
17		Right  front  top corner of the room.
18		In side the (LAF 1) left rear top corner.
19		In side the (LAF 1) right rear top corner.
20		In side the (LAF 2) left rear top corner.
21		In side the (LAF 2) right rear top corner.
22	Return Chane Room	Top corner of the room near the change room -1 door

·         Perform the fogging as per procedure.

·         Record the fogging details.

·         After completion of fogging  and defogging enter the area and take the exposed biological indicators  SS coupons / self contained biological indicators.

·         Clean the complete area as per procedure.

·         Analyse the biological indicators as per below mentioned procedure.

1.    SS coupons :

·         Take the required number of sterile 100 ml SCDM bottles  transfer in to 10 ml sterile test tubes.

·         Aseptically peel out the wrapper of the exposed biological indicators and transfer the indicator disk to the 10 ml test tube.

·         Transfer one un exposed biological indicator in the same manner and considered as  a positive control.

·         And one 10 ml Scdm tube considered as a negative control.

·         Incubate the test samples at 55⁰ to 60⁰ C for 7 days .

·         Observe the results on each working day.                                                                                                        

2.   Self –Contained Biological Indicators :

·         after exposer crush the biological indicators aseptically.

·         Incubate the test samples at 55⁰ to 60⁰ C for 1 or 2 days  (based on manufactures recommendation).

·         Observe the results on each working day.

·         Record the analysis and observations.

·         If growth observed within the incubation period , record the results accordingly.

·         Decontaminate the material after completion of observation.
        Any discrepancy and corrective action are observed in validation shall be reported.

Acceptance Criteria:

·         Test sample : No bacterial growth should be observed  during the incubation at 55⁰ to 60⁰ C for exposed indicators.

·         Positive Control :Growth should be observed  during the incubation at 55⁰ to 60⁰ C for Un-exposed indicators.

·         Negative Control : No bacterial growth should be observed  during the incubation at 55⁰ to 60⁰ C for 10 ml SCDM tube.

Conclusion :

·         6 % Hydrogen peroxide (H₂O₂)can effectively control the microbial growth  and it can be used a fogging solution for regular fogging activity of controlled environment.