Preparation

a. Cleaning: Remove gross filth and visible soil prior to application. Wash soiled surfaces with a compatible detergent using a cloth, sponge or appropriate cleaning device to ensure visible soils are removed. Rinse with potable water and allow to air dry. All the surfaces in the treatment area must be completely dry to the touch or visibly dry prior to vHP application.

b. The vHP Application Equipment: Position or connect the vHP application equipment for optimum vHP distribution into the treatment enclosure. See Equipment User’s Manual for proper equipment preparation and setup.

c. Sealing: Seal the treatment enclosure adequately to assure that hydrogen peroxide levels outside the enclosure are kept at acceptable levels [< one ppm tune weighted average for eight hours (TWA)] and ensure sufficient concentration of vHP Biostat in the treatment enclosure.

a. Close and seal windows and doors (if required). Sealing techniques can vary, but most often includes polyethylene sheeting and adhesive tape. Verify effectiveness of the sealing process by conducting an air draft potential analysis using a smoke stick test to ensure there are no leaks where openings have been sealed in the enclosure.
b. Turn off all ventilation systems including HVAC and seal any supply or return vents/ductwork.
c. Monitor areas immediately adjacent to the fumigated space to ensure levels are below TWA for hydrogen peroxide.

d. Securing Enclosure:

i. Assure all personnel have vacated the treatment enclosure prior to vHP application. Remove all plants, animals, beverages and food.
ii. Applicators must not reenter the treated enclosure until exposure levels of hydrogen peroxide are at/or
iii. below one ppm.

e. Placarding of Treatment Enclosure: The applicator must placard or post all entrances to the treatment enclosure and designated buffer zones with signs in English bearing:

a. The signal word “DANGER/PELIGRO” in red.
b. “Area under treatment, “DO NOT ENTER/NO ENTER.
c. The statement “This sign may only be removed after the treatment enclosure has been aerated to hydrogen peroxide levels less than or equal to one ppm”.
d. Identification of hydrogen peroxide as hazard associated with the treatment process.
e. Contact information for the applicator.

All entrances to the treatment enclosure must be placarded. Placards must be placed in advance of the treatment in
order to keep unauthorized persons from entering the treated enclosure. Placards are removed after the treatment
enclosure contains concentrations of hydrogen peroxide at/or below one ppm.

Application:

BioClean vHP Hydrogen Peroxide Biostat may also be applied to dry, sealed pre-cleaned enclosures without prior validation when the area is treated on a non-routine basis or enclosures being treated vary in configuration, materials of composition and content of items located in the treatment enclosure. The use of the vHP process in these conditions requires the applicator to apply a fixed vHP concentration over a set contact time. In addition the enclosure must be dehumidified and conditioned as part of the application process and aerated after TREATMENT. BioClean vHP Biostat may be applied at a set concentration and contact time to sealed enclosures of up to 4,000 ft3 in industrial, commercial and institutional settings (including production operations in pharmaceutical manufacturing, manufacturing clean rooms, medical device TREATMENT as part of a manufacturing process, laboratories, animal research facilities, patient rooms, hotel rooms, offices, cruise ships, recreational facilities and emergency response vehicles). In these applications, the vHP concentration should be monitored using a hydrogen peroxide sensor to ensure an adequate concentration level is maintained during the TREATMENT phase of the process. In addition, hydrogen peroxide chemical indicators must be placed throughout the enclosure to be treated to verify distribution of hydrogen peroxide throughout the enclosure. If more than one room of a consistent dimension is being treated, the applicator
may use the same vHP cycle settings as established in the initial room without use of a vHP sensor to confirm the concentration of the treatment cycle. These operations should be carried out by Huguenot Laboratories trained and certified applicators familiar with the set up and operation of vHP application equipment.

Treatment of Sealed, Dry Precleaned Enclosures at 250 ppm BioClean vHP Biostat for 90 minutes:

Prepare the treatment enclosure as defined above (Preparation of Enclosures Section) including pre-cleaning, drying and preparation of vHP Generator (refer to User’s Manual for vHP Generating Unit), sealing the enclosure and placarding of the enclosure to be treated. Place vHP monitor in a location most difficult for vHP target concentration to be reached in the treatment enclosure. This is typically in a comer in the enclosure farthest away from the vHP generation unit. All drawers, closets & cabinet doors, etc. must be opened to permit exposure to vHP Biostat. Place chemical indicators throughout the enclosure to verify effective distribution of vHP Biostat. The number of indicators placed throughout the enclosure must be based on the formula of one chemical indicator per 100 ft2. The chemical indicators must be placed in room corners and in areas difficult for the vHP Biostat to access such as closets, dressers, cabinets or other partially occluded areas. Place oscillating fans throughout the enclosure to facilitate effective distribution of the vHP Biostat. Program the vHP Generator to initiate a DEHUMIDIFICATION phase to achieve < 70% relative humidity. Assure the ambient temperature is not less than 2 1° C or 70° F initially and throughout the fumigation process. Once the DEHUMIDIFICATION phase is complete initiate a CONDITIONING phase to achieve a 250 ppm vHP Biostat concentration in the sealed enclosure. When a 250 ppm vHP Biostat concentration is achieved initiate the TREATMENT phase and maintain this concentration for 90 minutes. During the TREATMENT phase, monitor areas adjacent to the sealed enclosure with devices such as Drager tubes to assure hydrogen peroxide levels do not exceed one ppm. If this level is exceeded outside the treatment enclosure, the applicator should immediately abort the treatment process and ensure the enclosure is properly sealed. Upon completion of the TREATMENT phase, begin the AERATION phase to reduce levels of hydrogen peroxide at or below one ppm (TWA).

Treatment of Sealed, Dry Precleaned Enclosures at 400 ppm BioClean vHP Biostat for 30 minutes:

Prepare the treatment enclosure as defined above (Preparation of Enclosures Section) including pre-cleaning, drying and preparation of vHP Generator (refer to User’s Manual for vHP Generating Unit), sealing the enclosure and placarding of the enclosure to be treated. Place vHP monitor in a location most difficult for vHP target concentration to be reached in the treatment enclosure. This is typically in a corner in the enclosure farthest away from the vHP generation unit. All drawers, closets & cabinet doors, etc. must be opened to permit exposure to vHP Biostat. Place chemical indicators throughout the enclosure to verify effective distribution of vHP Biostat. The number of indicators placed throughout the enclosure must be based on the formula of one chemical indicator per 100 ft2. The chemical indicators must be placed in room corners and in areas difficult for the vHP Biostat to access such as closets, dressers, cabinets or other partially occluded areas. Place oscillating fans throughout the enclosure to facilitate effective distribution of the vHP Biostat. Program the vHP Generator to initiate a DEHUMIDIFICATION phase to achieve < 70% relative humidity. Assure the ambient temperature is not less than 21 C or 70 F initially and throughout the fumigation process. Once the DEHUMIDIFICATION phase is complete initiate a CONDITIONING phase to achieve a 400 ppm vHP Biostat concentration in the sealed enclosure. When a 400 ppm vHP Biostat concentration is achieved initiate the TREATMENT phase and maintain this concentration for 30 minutes. During the TREATMENT phase, monitor areas adjacent to the sealed enclosure with devices such as Drager tubes to assure hydrogen peroxide levels do not exceed one ppm. If this level is exceeded outside the treatment enclosure, the applicator should immediately abort the treatment process and ensure the enclosure is properly sealed. Upon completion of the TREATMENT phase, begin the AERATION phase to reduce levels of hydrogen peroxide at or below one ppm (TWA).

Monitoring of H2O2 Concentrations in the Sealed Enclosure and Reentry Instructions Following Aeration.

vHP Monitoring: Drager tubes or other vHP monitoring devices are utilized by means of a minimally invasive
technique for vHP sampling to determine the vHP concentration in the sealed enclosure during and after the
aeration phase. After the vHP concentration within the treated enclosure is at or below the OSHA Permissible
Exposure Limit (PEL) of one ppm, the enclosure may be released to normal operations and general public use.

Criteria for Successful Fumigation:

1. All vHP fumigation process conditions (vapor concentration, temperature, relative humidity are achieved throughout the fumigation cycle.
2. ATP Testing is Complete: ATP is a molecule found in and around living cells, and as such it gives a direct measure of biological concentration and health. ATP is quantified by measuring the light produced through its reaction with the naturally occurring firefly enzyme luciferase using a luminometer. The amount of light produced is directly proportional to the amount of ATP present in the sample.

Reentry Instructions:

1. Early reentry in the case of an emergency requires wearing a Self Contained Breathing Apparatus (SCBA)
operated in pressure-demand mode, full hydrogen peroxide resistant body suit, gloves and boots to protect from the inhalation hazard as well as the corrosive action of hydrogen peroxide to tissues. When entering into the area under fumigation always work with two or more people under the direct supervision of a trained applicator wearing appropriate respirators.

2. Reentry to the sealed enclosure by a trained and certified applicator is allowed with a self contained breathing apparatus at vHP concentrations up to 5 ppm to allow for windows to be opened and to augment the aeration process if deemed appropriate at the specific location by the trained and certified applicator. Otherwise, do not reenter the treated enclosure until exposure levels of hydrogen peroxide are at or below one ppm.

Releasing Treated Sealed Enclosure for Return to Service:

a. Once vHP levels are determined to be at or below one ppm, applicators may re-enter the treated enclosure and remove any sealing materials and disconnect/remove vHP Generator from the treated sealed enclosure.
b. Turn on ventilation systems including HVAC.
c. Remove placards and release the treated enclosure for normal operation and use after the levels of hydrogen peroxide are determined to be at or below one ppm.
d. Release the treated enclosure for general public use after hydrogen peroxide levels are determined to be at or below one ppm.