[Title 32 CFR E]
[Code of Federal Regulations (annual edition) - July 1, 2002 Edition]
[Title 32 - NATIONAL DEFENSE]
[Chapter V - DEPARTMENT OF THE ARMY]
[Subchapter H - SUPPLIES AND EQUIPMENT]
[Part 627 - THE BIOLOGICAL DEFENSE SAFETY PROGRAM, TECHNICAL SAFETY REQUIREMENTS (DA PAMPHLET 385-69)]
[Subpart E - Decontamination and Disposal]
[From the U.S. Government Printing Office]


32NATIONAL DEFENSE32002-07-012002-07-01falseDecontamination and DisposalESubpart ENATIONAL DEFENSEDEPARTMENT OF THE ARMYSUPPLIES AND EQUIPMENTTHE BIOLOGICAL DEFENSE SAFETY PROGRAM, TECHNICAL SAFETY REQUIREMENTS (DA PAMPHLET 385-69)
                 Subpart E--Decontamination and Disposal



Sec. 627.32  Introduction.

    All material or equipment that is potentially contaminated with 
etiologic agents must be rendered nonhazardous before disposal. This 
chapter describes the acceptable physical and chemical decontamination 
methods and the general applicability of each. In general, all 
infectious materials and all contaminated equipment or apparatus will be 
sterilized before being washed and stored or discarded.



Sec. 627.33  Methods of decontamination.

    (a) Autoclave. The use of wet heat is the most dependable procedure 
for destroying all forms of microbial life. An autoclave employs 
saturated steam under a pressure of approximately 15

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pounds per square inch (psi) to achieve a chamber temperature of at 
least 121  deg.C for a minimum of 15 minutes. The time is measured after 
the temperature of the material being sterilized reaches 121  deg.C. 
Other combinations of temperature and pressure (some of which are 
dependent on the equipment used) can be used to accomplish sterilization 
provided that the efficacy of sterilization is validated as described 
below. The most critical factor in ensuring the reliability of this 
sterilization method, other than proper temperature, is preventing 
entrapped air that is not replaced by stem. Material to be autoclaved 
must come in contact with steam and heat and, as a result, it may be 
necessary to add water to a load of waste to aid in the formation and 
penetration of steam. Autoclaves use either a steam-activated exhaust 
valve that reamins open during the replacement of air by live steam 
until the steam triggers the valve to close, or a pre-cycle vacuum to 
remove air prior to steam introduction.
    (b) Sterilization will be verified using biological indicators (for 
example, Bacillus stearothermophilus spores) at locations throughout the 
autocalve, to include placement in the center of test loads, when the 
autoclave is first put into service, and after any maintenance or 
repairs. The primary means of verifying routine sterilization will be 
through using chemical indicators (for example, autoclave tape or 
labels) at locations throughout the autoclave. In addition each 
autoclave will be equipped with a permanent means to record time and the 
temperature of each operational event as a means of ensuring 
sterilization. The type of materials being handled must be reviewed and 
standard conditions for sterilization of each established. As a guide, 
the manufacturer's manual for the autoclaves will be consulted as a 
starting point in establishing these conditions. Treatment conditions to 
achieve sterility will vary in relation to the volume of material 
treated, the contamination level, the moisture content, and other 
factors that should be considered and which may cause the times to 
lengthen. In each case, the conditions will be established based on 
tests which verify that the conditions selected are effective. In 
addition to being effective from viable agents, autoclaving effectively 
inactivates most protein toxins.
    (c) Dry heat. Dry heat requires longer times or higher temperatures 
or both than does wet heat. If used, the specific sterilization times 
and temperatures must be determined for each type of material being 
sterilized. In general, sterilization by dry heat can be accomplished at 
169-170  deg.C for periods of 2 to 4 hours. Higher temperatures reduce 
the time requirements. The heat transfer properties and spatial relation 
or arrangement of materials in the load are critical in ensuring 
effective sterilization.
    (d) Liquid disinfectants. Liquid disinfectants may be used in 
surface treatment, in dip tanks, and, at sufficient concentration, as 
sterilants of liquid waste for final disposal. If liquid disinfectants 
are used, they must have been shown to be effective against the 
organisms present. Important considerations include: temperature, time 
of contact, the negative logarithm of hydrogen ion concentration (pH), 
concentration and state of dispersion, penetrability, and reactivity of 
organic material at the site of application. Small variations in these 
factors may make large differences in the effectiveness of disinfection, 
so complete reliance should not be placed on liquid disinfectants when 
the end result must be sterility. If evidence of efficacy under the 
proposed procedures has not been reported previously, preliminary 
studies to verify the efficacy of liquid disinfectants must be 
conducted. Such studies may include attempts to recover and quantitate 
the agent in question from liquid or swab samples, or sealed patches, by 
animal inoculation, plaque assay, agar or broth cultivation, and similar 
methods, following controlled decontamination under the same 
experimental conditions envisioned for the proposed studies.
    (1) Alcohol. Ethyl or isopropyl alcohol at the concentration of 70-
85 percent by weight will denature proteins but is slow in its 
germicidal action. Alcohols are effective disinfectants for lipid-
containing viruses. These alcohols exhibit no activity against bacterial 
spores.

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    (2) Phenolic compounds. These are effective disinfectants against 
vegetative bacteria, including Mycobacterium tuberculosis, fungi, and 
lipid-containing viruses. The phenolics are not effective against 
bacterial spores or non-lipid-containing viruses. The concentrations 
used will be in accordance with the manufacturer's recommendations.
    (3) Formaldehyde solutions. Formaldehyde in solution at a 
concentration of 8 percent (formalin) is effective against vegetative 
bacteria, spores, and viruses. It loses considerable disinfectant 
activity below room temperature. Due to the toxic properties of 
formaldehyde, the use of formalin is restricted to surfaces or materials 
that are contained within appropriate engineering controls.
    (4) Quaternary ammonium compounds. These cationic detergents are 
strongly surface-active. They lose effectiveness in the presence of 
proteins and are neutralized by anionic detergents, such as soap. At low 
concentrations, they are bacteriostatic, tuberculostatic, sporostatic, 
fungistatic, and algistatic. At medium concentration, they are 
bactericidal, fungicidal, algicidal, and virucidal against lipophilic 
viruses. They are not tuberculocidal, sporicidal, or virucidal against 
hydrophilic viruses, even at high concentrations. The manufacturer's 
recommended dilution will be used.
    (5) Chlorine. Sodium hypochlorite is normally used as a base for 
chlorine disinfectants. Free available chlorine is the active ingredient 
and, at concentrations of at least 2,500 parts per million (ppm) (0.25 
percent), is a disinfectant that is active against most microorganisms 
and bacterial spores. Chlorine solutions at 2.5 percent free available 
chlorine are effective against most toxins. Chlorine solutions lose 
strength if exposed to air, so fresh solutions must be prepared whenever 
the free chlorine content falls below desired minimums.
    (6) Iodine. The characteristics of chlorine and iodine are similar. 
Iodophor compounds with 1,600 ppm free available iodine provide a 
relatively rapid inactivation of all microorganisms, including some 
bacterial spores. A commonly available iodophor is Wescodyne. The 
manufacturer of Wescodyne recommends a range of dilution form 1 to 3 
ounces per 5 gallons of water, giving a solution containing from 25 to 
75 ppm of free iodine. At these concentrations, available iodine may be 
rapidly taken up by any extraneous protein present and will not be an 
effective sporocide. A solution providing 1,600 ppm iodine is 
recommended for hand washing or for use as a sporocide.
    (7) Mercurials. Although the mercurials exhibit good activity 
against viruses, they are toxic and are not recommended for general use. 
They have poor activity against vegetative bacteria and are totally 
ineffective sporicides. The dilution recommendations stated by the 
manufacturer will be followed.
    (e) Vapors and gases. Formaldehyde, ethylene oxide, peracetic acid, 
beta-propiolactone, methyl bromide, and glutaraldehyde have all been 
used successfully as space sterilants where they can be employed in 
closed systems and with controlled conditions of temperature and 
humidity. Of these, methyl bromide, beta-propiolactone, and 
glutaraldehyde are not recommended because of their toxic properties. 
Peracetic acid can readily decompose with explosive violence in a 
concentrated state and must be used only in a diluted state and with 
extreme care. Formaldehyde and ethylene oxide are both regulated by OSHA 
for their potential human carcinogenicity, but do have permissible 
exposure levels (unlike beta-propiolactone, for example) and can be used 
safely under controlled conditions.
    (1) Formaldehyde. Formaldehyde gas is, in general, the chemical of 
choice for space disinfection. Biological safety cabinets and associated 
effluent air-handling systems and air filters, incubators, laboratory 
rooms, buildings, or other enclosed spaces can be disinfected with 
formaldehyde. The procedures found in appendix E of the National 
Sanitation Foundation Standard Number 49 will be followed for the 
disinfection of biological safety cabinets. Other enclosures or areas 
will be disinfected by following the same principles. To disinfect 
rooms, the generation of formaldehyde gas from heating powdered or flake 
paraformaldehyde is

[[Page 558]]

the preferred method. When area decontamination is performed, use 0.3 
grams of paraformaldehyde for each cubic foot of space to be treated. 
The room or area must be above 70  deg.F, the relative humidity above 70 
percent, and the exposure time at least 2 hours (overnight is 
preferred). After the required time for disinfection, the room must be 
cleared of the formaldehyde gas (a small room with nonporous surfaces 
and no materials or equipment in the room can be cleared of all 
detectable formaldehyde by aeration for one hour, while larger areas 
with equipment in them may take a full day). Before formaldehyde is used 
as a space disinfectant, the area to be treated must be surveyed to 
ensure that there are no open containers of any acidic solution 
containing chloride ion in order to prevent the possible formation of 
bis (chloromethyl)ether, a human carcinogen. Specific OSHA requirements 
for posting of rooms and equipment, personnel protection, and other 
requirements are found in 29 CFR 1910.1048.
    (2) Ethylene oxide (EtO). EtO sterilization will only be conducted 
in a sterilizer designed for that purpose and designed to maintain 
potential exposure levels below the current OSHA standard. EtO is 
effective against all microorganisms, including spores, molds, 
pathogenic fungi, and highly resistant thermophilic bacteria. All 
materials to be used in contact with human skin (for example, clothing, 
shoes, masks, adhesive tape) must be aerated for at least 24 hours after 
sterilization and prior to use. Concentrations of 500 to 1000 ppm are 
required for sterilization. Specific OSHA requirements for the use of 
ethylene oxide are found in 29 CFR 1910.1047.
    (f) UV Radiation. UV radiation at a wave length of 253.7 nanometers 
is a practical method for inactivating airborne virsuses, mycoplasma, 
bacteria, and fungi. The usefulness of UV radiation on exposed surfaces 
is limited by its low penetrating power. UV radiation shall only be 
relied upon to sterilize surfaces when conventional methods, such as 
autoclaving or the use of liquid disinfectants, would make the product 
unusable. An example is data sheets that must be brought out of a 
biocontainment facility. The UV intensity must be at least 40 
microwatts/cm 3 on the surface to be treated. Single sheets 
of paper may be treated by exposing them to this radiation for a minimum 
of 15 minutes. A calibrated photoelectric UV intensity meter, capable of 
measuring UV radiation at a wave length of 253.7 nanometers, will be 
used whenever a new UV source is installed, and quarterly thereafter, to 
ensure the UV source is providing at least 40 microwatts/cm 3 
at the work surface. Bulbs should be cleaned routinely to remove any 
accumulated dust and prolong bulb performance and assure proper energy 
output. Protective eye wear and clothing may be necessary when working 
around UV radiation.



Sec. 627.34  Disposal.

    Inactivation is the first step in the disposal of etiologic agents 
or materials that are potentially contaminated with them. All 
contaminated or potentially contaminated materials must be effectively 
disinfected or sterilized by an approved procedure discussed in 
Sec. 627.33. After decontamination, reusable items, such as clothing or 
glassware, may be washed with other uncontaminated or decontaminated 
items.
    (a) Combustible items. Combustible disposable items should be bagged 
and incinerated in an appropriate approved incinerator or otherwise 
disposed of in accordance with State and local regulations.
    (b) Noncombustible disposable items. Items will be packaged as 
stated in Sec. 626.34(e) and disposed of by a licensed waste hauler.
    (c) Equipment. Equipment that cannot be autoclaved will be 
decontaminated by gaseous sterilization or with a suitable liquid 
disinfectant. Such equipment will be certified as decontaminated by the 
safety officer.
    (d) Waste. Materials generated, such as solvents, acids, chemical 
carcinogens, radioactive isotopes, medical waste, or dead animals must 
be decontaminated, packaged, and then disposed of in accordance with 
EPA, NRC, local, State, and Federal regulations.

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    (e) Mixed waste. When two or more hazardous materials are mixed 
together, the mixture will be decontaminated and disposed of in 
accordance with EPA, NRC, State, and Federal regulations for the 
mixture, or for the most hazardous material.
    (f) Packaging. Solid waste will be placed in cans, sturdy bags, or 
boxes. Rigid, puncture-resistant, sealable containers will be used for 
packaging ``sharps.'' When wet materials are packaged for disposal, the 
materials will be placed in a leak-proof container. Heavy waste will be 
placed in rigid containers ensuring that the burst strength of the 
container is not exceeded.
    (g) Labeling. A method of verifying that all items prepared for 
disposal have been decontaminated will be established for etiologic 
agent wastes. Mixed waste will be labeled as appropriate to indicate the 
hazards that must be addressed after decontamination.
    (h) Recordkeeping. A manifest will be initiated and maintained, 
where required, to record the disposition and transfer of waste. 
Applicable Federal, State, and local ordnances will be followed.