fiLi'i United States
Environmental Protection
^^1 i^lfcAgency
Verification of Hydrogen Peroxide Vapor
Technologies for Decontaminating Indoor
Surfaces Contaminated With Biological or
Chemical Agents
Office of Research arid Development
National Homeland Security
Research Center
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TEST/QUALITY ASSURANCE PLAN
for
VERIFICATION OF HYDROGEN PEROXIDE VAPOR TECHNOLOGIES
FOR DECONTAMINATING INDOOR SURFACES CONTAMINATED
WITH BIOLOGICAL OR CHEMICAL AGENTS
Prepared by
Battelle
Columbus, Ohio
GSA Contract Number GS-23F-0011L
Blanket Purchase Agreement 2C-R903-NBLX
Task Order Number 1103
EPA Task Order Project Officer
John C.S. Chang
July 21, 2003
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TABLE OF CONTENTS
Page
1.0 BACKGROUND 1
1.1 T echnology V erification 1
1.2 Test Obj ective 2
1.3 Organization and Responsibilities 2
1.3.1 Battelle 2
1.3.2 Vendor 5
1.3.3 EPA 5
1.3.4 Stakeholders 6
2.0 APPLICABILITY 7
2.1 Subject 7
2.2 Scope 8
3.0 TEST SITE 10
3.1 Site Description 10
3.2 Site Operations 12
4.0 EXPERIMENTAL DESIGN 14
4.1 General Test Design 14
4.1.1 Parameters to Be Tested 14
4.1.2 Scale of Testing 15
4.1.3 Efficacy 15
4.1.4 Temperature and Relative Humidity Conditions 16
4.1.5 Surface Damage 16
4.2 Agents and Surrogates 17
4.2.1 Chemical Agents and Surrogates 17
4.2.2 Biological Agents and Surrogates 18
4.3 Test Surfaces 19
4.4 Test Sequence 20
5.0 EQUIPMENT AND MATERIALS 21
5.1 Materials 21
5.1.1 Agents 21
5.1.2 Spore Strips 22
5.1.3 Surfaces to Be Tested 22
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TABLE OF CONTENTS (Continued)
5.2 Delivery and Application Equipment 22
5.2.1 Agent/Surrogate Surface Application 22
5.2.2 Temperature/Humidity Conditions 22
5.3 Test Chamber 23
5.4 Sampling and Analysis Materials and Equipment 25
5.4.1 Chemical Agent Testing 25
5.4.2 Biological Agent Testing 27
5.5 Performance Evaluation Audit Materials 27
6.0 TEST PROCEDURES 28
6.1 Method Demonstration 28
6.1.1 Chemical Agent Method Demonstration 28
6.1.2 Biological Agent Method Demonstration 29
6.2 Coupon-Scale Testing 29
6.2.1 Preparation of Test Materials 30
6.2.2 Application of Agents to Test Coupons 31
6.2.3 Confirmation of Surface Applications 32
6.2.4 Application of Hydrogen Peroxide Vapor
Decontamination Technology 32
6.2.5 Determination of Decontamination Efficacy 33
6.2.6 Observation of Surface Damage 38
7.0 QUALITY ASSURANCE/QUALITY CONTROL 39
7.1 Equipment Calibrations 39
7.2 Assessments and Audits 40
7.2.1 Technical Systems Audits 40
7.2.2 Performance Evaluation Audit 40
7.2.3 Data Quality Audit 41
7.2.4 Assessment Reports 41
7.2.5 Corrective Action 42
8.0 DATA ANALYSIS AND REPORTING 43
8.1 Data Acquisition 43
8.2 Calculation Procedures 44
8.2.1 Data Screening 44
8.2.2 Efficacy 45
8.2.3 Contact Transfer 46
8.2.4 Offgas Flux 47
8.3 Data Review 47
8.4 Reporting 48
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TABLE OF CONTENTS (Continued)
9.0 HEALTH AND SAFETY 49
9.1 Access 49
9.2 Potential Hazards 49
9.3 Training 49
9.4 Safe Work Practices 50
10.0REFERENCES 51
APPENDIX A
TEST PERFORMANCE CONTROL SHEET/TEST COUPON SAMPLE FORM
APPENDIX B
VENDOR-SPECIFIC TEST INFORMATION
LIST OF FIGURES
Figure 1-1. Organization Chart for Hydrogen Peroxide Vapor Decontamination
Technology Verification Test 3
Figure 5-1. Compact Glove Box—BW Agent Tests 24
Figure 5-2. Contact Transfer Weights 25
Figure 5-3. Offgas Rack 26
Figure 5-4. Offgas Cell 26
LIST OF TABLES
Table 3-1. Certifications of HMRC and MREF 13
Table 4-1. Chemical Agents to Be Used 17
Table 4-2. Sequence of Test Procedures in Verification Testing of
Hydrogen Peroxide Vapor Decontamination Technologies 20
Table 7-1. Calibration Ranges 40
Table 8-1. Summary of Data Recording Process for Verification Testing 44
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List of Acronyms
AAALAC American Association for the Accreditation of Laboratory Animal Care
AC hydrogen cyanide
BSCs biosafety cabinets
BW biological warfare
BWD bare wood
CASARM Chemical Agent Standard Analytical Reference Material
CB chemical and biological
CG phosgene
CK cyanogen chloride
CoC chain-of-custody
CSM Chemical Surety Material
CT contact transfer
CV coefficient of variation
CW chemical warfare
DEMTMP diethylmethylthio-methylphosphate
DL decorative laminate
E efficacy
EPA U.S. Environmental Protection Agency
ETV Environmental Technology Verification
FID flame ionization detectors
FPD flame photometric detectors
FSP Facility Safety Plan
GA tabun
GB sarin
GC gas chromatograph
GD soman
GM galvanized metal ductwork
GS glass
H2O2 hydrogen peroxide
HOD sulfur mustard
HL mustard-lewisite mixtures
HML Hazardous Materials Laboratory
HMRC Hazardous Materials Research Center
HPV Hydrogen peroxide vapor
IACUC Institutional Animal Care and Use Committee
IS internal standard
L Lewisite
LC/MS/MS liquid chromatograph/mass spectrometry
LITF Large Item Test Facility
MREF Medical Research and Evaluation Facility
MS mass spectrometry
MSDS Material Safety Data Sheets
NHSRC National Homeland Security Research Center
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List of Acronyms (Continued)
OF
offgas flux
ORI
Operational Readiness Inspection
PA
peak area
PC
painted concrete
PE
performance evaluation
PFIB
perfluoroisobutylene
PPE
personal protective equipment
PS
painted steel
PW
painted wallboard
QA
quality assurance
QMP
Quality Management Plan
RDS
Research Dilute Solutions
RDT&E
research, development, test, and evaluation
SA
arsine
SBCCOM
Soldier Biological and Chemical Command
SCOB
Safety and Chemical Operations Branch
SD
standard deviation
SOPs
standard operating procedures
TEP
triethyl phosphate
TGD
thickened GD
THD
thickened HD
TICs
toxic industrial chemicals
TOPO
Task Order Project Officer
TPCSs
Test Performance Control Sheets
TSA
technical systems audit
TSA
Trypticase Soy Agar
USAMRMC
United States of America Medical Research Material Command
vx
V-agent
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DISTRIBUTION LIST
Dr. Thomas J. Kelly
Battelle
505 King Avenue
Columbus, Ohio 43201-2693
Biju Stephen
BIOQUELL, Inc.
34 Walworth Road
Andover, Hampshire SP10 5AA
United Kingdom
Ms. Karen Riggs
Battelle
505 King Avenue
Columbus, Ohio 43201-2693
Mr. Zachary Willenberg
Battelle
505 King Avenue
Columbus, Ohio 43201-2693
Ms. Shirley Wasson
U.S. Environmental Protection Agency
Mail Drop E343-03
Research Triangle Park, NC 27711
Dr. John C.S. Chang
U.S. Environmental Protection Agency
U.S. EPA Mailroom, E305-03
Research Triangle Park, NC 27711
Ms. Carol Sabourin
Battelle
505 King Avenue
Columbus, Ohio 43201-2693
Mr. Daniel S. Janke
Battelle
505 King Avenue
Columbus, Ohio 43201-2693
Mr. Gary Stickel
Battelle
505 King Avenue
Columbus, Ohio 43201-2693
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EPA/Battelle Approval of EPA/ETV Test/QA Plan for
Verification of Hydrogen Peroxide Vapor
Technologies for Decontaminating Indoor Surfaces
July 2003
(SIGNA TURES ON FILE)
Original signed by:
Original signed by:
John Chang
7/22/03
John Chang, Ph.D.
Task Order Project Officer
U.S. EPA
Date
Shirley Wasson
Shirley Wasson
Quality Manager
U.S. EPA
7/22/03
Date
Original signed by:
Original signed by:
Karen Riggs
Zachary Willenberg 7/22/03
7/23/03
Karen Riggs Date Zachary Willenberg Date
Program Manager Quality Manager
Battelle Battelle
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Vendor Approval of EPA/ETV Test/QA Plan for
Verification of Hydrogen Peroxide Vapor
Technologies for Decontaminating Indoor Surfaces
Contaminated with Biological or Chemical Agents
July 2003
(SIGNA TURE ON FILE)
Name BiJu Stephen, Operation Mgr Signature Biju Stephen
Company BIOQUELL, Inc.
Date 7/24/03
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1.0 BACKGROUND
1.1 Technology Verification
Among its responsibilities related to Homeland Security, the U.S. Environmental Protection
Agency (EPA) has the goal of identifying methods and equipment that can be used for
decontaminating indoor environments, following a terrorist attack on a building using chemical
or biological agents. In January 2003, EPA established the National Homeland Security
Research Center (NHSRC) to manage, coordinate, and support a wide variety of homeland
security research and technical assistance efforts. The Safe Buildings Program, a key research
component of the NHSRC, has the aim of verifying the performance of products, methods, and
equipment that can decontaminate chemical or biological agents on indoor surfaces or in indoor
air.
To accomplish this aim, the EPA has expanded the scope of its Environmental Technology
Verification (ETV) program. The ETV process, which has been used since 1997 to verify the
performance of over 200 environmental technologies, includes developing a test/quality
assurance (QA) plan with input from stakeholders and vendors; applying high-quality test
procedures according to that plan; and publicizing separate performance reports for each
technology verified. The ETV process does not rank, select, or approve technologies, but instead
provides credible performance data to potential users and buyers. Other information about the
program is available at the ETV web site (http://www.epa.gov/etv) and through the NHSRC web
site (www.epa.gov/nhsrc).
In expanding the ETV program to address Homeland Security needs, the EPA established the
ETV Building Decontamination Technology Center, which is managed by Battelle, of
Columbus, OH, under contract with EPA. Verification testing of decontamination technologies
in the Center generates objective performance data so building and facility managers, first
responders, groups responsible for building decontamination, and other technology buyers and
users can make informed purchase and application decisions. Verification tests are conducted in
the Center in accordance with the ETV process, under the direction of the EPA. All verification
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activities are subject to the Quality Management Plan(1) and the Generic Verification Protocol(2)
for the Center. In performing each verification test, Battelle follows the procedures described in
those documents, and develops a separate test/QA plan appropriate for the decontamination
technology being tested. This document is the test/QA plan for verification testing of
decontamination technologies that use hydrogen peroxide vapor (HPV) as the decontaminating
agent.
1.2 Test Objective
The objective of this test/QA plan is to establish laboratory test procedures to determine the
efficacy of hydrogen peroxide vapor decontamination technologies, for removing or inactivating
chemical and biological agents and surrogates on a range of representative indoor surfaces.
1.3 Organization and Responsibilities
Verification testing under this test/QA plan will be performed by Battelle under the direction of
EPA, with input from expert stakeholders and decontamination technology vendors. The
organization chart in Figure 1-1 shows the organizations and individuals who will have
responsibilities under this plan. The responsibilities of these organizations and individuals are
summarized in the following subsections. Specific responsibilities are detailed for the Battelle
Test Coordinator, the technology vendor, and the test leaders as those most involved in
conducting the verification testing.
1.3.1 Battelle
Dr. Thomas J. Kelly is the Verification Testing Leader for the ETV Building Decontamination
Technology Center. He will have overall responsibility for ensuring that the technical, schedule,
and cost goals established for verification testing are met, and that the verification process
employed for testing is consistent with Center and ETV program guidelines. For this test, Dr.
Kelly will serve as the interface for the Center stakeholder committee.
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Figure 1-1. Organization Chart for Hydrogen Peroxide Vapor
Decontamination Technology Verification Test
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Ms. Karen Riggs is Battelle's Manager for the contract under which the ETV Building
Decontamination Technology Center was established. Ms. Riggs will maintain communication
with EPA's Task Order Project Officer (TOPO) on all aspects of the program; monitor adherence
to budgets and schedules in this work; and ensure that necessary Battelle resources, including
staff and facilities, are committed to the verification test.
Mr. Zachary Willenberg is Battelle's. Quality Manager for the ETV Building Decontamination
Technology Center. He will review the draft test/QA plan, audit at least 10 percent of the
verification data, and ensure that all quality procedures specified in this test/QA plan and in the
QMP(1) are followed.
Mr. Daniel Janke is Battelle's Verification Test Coordinator for this test. His responsibilities
include:
• Selection of the appropriate facility or location for the testing
• Coordination with vendor representatives to facilitate the performance of testing
• Preparation of the draft test/QA plan, verification report, and verification statement
• Arrangements for use of the test facilities and establishment of test schedules
• Selection of qualified staff to conduct the tests
• Assurance that testing is conducted according to this test/QA plan
• Input into revision of the test/QA plan, verification report, and verification statement in
response to reviewers' comments
• Updating the Battelle Center Manager and Verification Testing Leader on progress and
difficulties in planning and conducting the test
• Coordination with the Battelle Quality Manager for the performance of technical and
performance audits as required by Battelle or EPA Quality Management staff.
The Chemical and Biological Test Facilities at Battelle will serve as the location for the testing
described in this test/QA plan. These facilities are described in Section 3 of this plan. Biological
testing will be led by Dr. Carol Sabourin; chemical testing will be led by Mr. Gary Stickel. In
general, the responsibilities of the test leaders will be to:
• Assist in planning and scheduling the testing
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• Become familiar with the use of the technology to be tested
• Ensure that the facility is fully functional prior to the times/dates needed for
verification testing
• Provide requisite technical staff during verification testing
• Provide any safety training needed by Battelle, vendor, or EPA staff
• Review and approve all data and records related to facility operation
• Adhere to the requirements of this test/QA plan and the QMP(1) in carrying out the
verification testing
• Provide input on facility procedures for the verification report
• Support Mr. Janke in responding to any issues raised in assessment reports and audits
related to facility operation.
1.3.2 Vendor
The decontamination technology vendor will:
• Provide input for preparation of the test/QA plan
• Review the draft test/QA plan, and approve the final version
• Sign a Vendor Agreement specifying the respective responsibilities of the vendor and of
Battelle in the verification testing
• Provide the necessary materials and equipment to implement the decontamination
technology for testing
• Train Battelle and/or test facility staff in the application of the decontamination
technology
• Provide support, if needed, in use of the technology during testing
• Review the draft verification report and verification statement resulting from testing.
1.3.3 EPA
Dr. John Chang is EPA's TOPO for the ETV Building Decontamination Technology Center.
As such, Dr. Chang will have overall responsibility for directing the verification process and
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Battelle's activities, and will oversee the EPA review process on the draft test/QA plan,
verification report, and verification statement.
Ms. Shirley Wasson is EPA's Quality Manager for the ETV Building Decontamination
Technology Center. Ms. Wasson will lead EPA's QA oversight on this verification, including, at
her option, one external technical systems audit during verification testing.
1.3.4 Stakeholders
A group of approximately 25 experts from the first responder community, federal and state
agencies, military agencies, and academia serves as volunteer advisors to the ETV Building
Decontamination Technology Center. Battelle Center staff communicate with these stakeholders
regularly by e-mail or telephone and meet periodically with the stakeholder committee and the
EPA TOPO. The responsibilities of assigned stakeholders from this committee for testing are to
provide input on test procedures for preparation of the test/QA plan, review the draft test/QA
plan, and serve as peer reviewers for the verification report.
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2.0 APPLICABILITY
2.1 Subject
This test/QA plan is applicable to verification testing of decontamination technologies that
generate hydrogen peroxide vapor to decontaminate indoor surfaces contaminated by chemical
or biological agents. This plan is specifically focused on decontamination of the building
environment, for purposes of restoring a public building to a usable state after a contamination
episode. Decontamination of personnel or equipment is not the subject of this test/QA plan.
The decontamination technologies to be tested under this plan are based on dispersion of
hydrogen peroxide vapor into indoor spaces. Because hydrogen peroxide vapor is not stable as a
compressed gas, it must be produced on site by vaporization of concentrated aqueous solutions
of hydrogen peroxide. Thus, these technologies include the equipment and chemicals for
generating and dispersing the hydrogen peroxide vapor. These technologies may require specific
temperature and humidity conditions that favor the effectiveness of the decontamination process,
and may include systems to achieve the proper conditions in the volume to be decontaminated.
The chemical and biological agents that may pose a threat in the building environment include
toxic industrial chemicals (TICs), chemical warfare (CW) agents, and biological warfare (BW)
agents (including biotoxins). The chemical and biological agents selected for use in testing were
chosen based on a brief threat summary(3) developed from general opinions of Battelle experts,
with additional input from Center stakeholders. In the context of decontamination, the
contaminants of interest for this plan are those that can persist on indoor surfaces, leading to
continuing chance of exposure long after the contamination occurs. Thus, highly volatile TICs
and CW agents are not included in testing under this plan because they can be readily removed
by ventilation of the building. By the same logic, a highly persistent biological contaminant
(anthrax spores) was selected for testing, as opposed to biological agents that cannot survive for
long after the contamination event.
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The indoor surfaces selected for testing under this plan represent those that must be
decontaminated to return a building to use, and do not include those that might simply be
removed from the building for disposal. Highly porous non-structural materials, such as ceiling
tiles, clothcovered furniture and cubicle walls, and draperies are among those that were deemed
likely to be removed from a building for disposal; consequently, those materials are not
considered as priority test substrates in this verification plan. Structural materials such as
wallboard, painted concrete, metal ductwork, and wood, and surfaces of furnishings, such as
laminate, are considered essential candidate substrates. Carpeting is also included, as a porous
material that could possibly be left in a building for decontamination.
Verification testing requires a basis for establishing the quantitative performance of the tested
technology. For the testing conducted under this test/QA plan, quantitative performance is
assessed primarily in terms of the efficacy of decontamination. For this assessment, sampling
and analysis methods are used to determine the extent of contamination before and after the use
of the decontamination technology.
2.2 Scope
The overall objective of the testing called for under this plan is to verify the efficacy of the
hydrogen peroxide vapor decontamination technologies, for selected chemical and biological
agents on representative indoor surfaces. Testing of each technology is to be conducted at
temperature and relative humidity conditions that would be appropriate for that technology in a
building undergoing decontamination.
The performance parameters to be evaluated in verification testing under this test/QA plan
include:
• Efficacy in destroying chemical agents and surrogates on selected indoor surfaces
• Efficacy in destroying biological agents and surrogates on the same indoor surfaces
• Generation of toxic degradation products from interaction of the decontaminant with
the target agents.
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Efficacy will be tested by applying chemical and biological agents and surrogates to test
surfaces, and comparing the residual contamination after use of the decontamination technology
to the contamination originally present. Generation of toxic degradation products will be
determined by analysis of the residual contamination for specific degradation products. In
addition, any apparent destructiveness of the decontaminant to test surfaces will be assessed by a
simple visual inspection before and after use of the decontamination technology.
Under this test/QA plan, verification of hydrogen peroxide vapor decontamination technologies
can include testing with both chemical and biological agents. These components of the complete
test are separate, and can be carried out at different times if necessary, and either the chemical or
biological agents can be excluded from testing if no efficacy is expected. If these components
are conducted separately, they may be the subjects of separate ETV verification reports.
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3.0 TEST SITE
Verification testing of hydrogen peroxide vapor decontamination technologies will be conducted
at Battelle's chemical and biological test facilities in West Jefferson, Ohio, near Battelle's
headquarters in Columbus, Ohio. The following sections describe the West Jefferson facilities.
The testing will be subject to facility-specific methods and SOPs as noted in this test/QA plan,
and are required for work at each facility. These documents are cited where appropriate
throughout this test/QA plan.
3.1 Site Description
Battelle's chemical and biological test facilities to be used for verification testing are:
• The Hazardous Materials Research Center (HMRC), a DoD laboratory-scale facility
conducting research with CW agents
• Medical Research and Evaluation Facility (MREF), which is a second DoD
laboratory-scale facility conducting research with CW and BW agents.
The HMRC is an ISO 9001 certified facility and provides a broad range of materials testing,
system and component evaluation, research and development, and analytical chemistry services
that require the safe use and storage of highly toxic substances. Since its initial certification by
the Chemical Research, Development and Engineering Center in 1981, the facility has
functioned as both a research and a technology development laboratory in support of DoD
chemical and biological (CB) programs. The HMRC and its personnel have the demonstrated
capability for storing and safely handling BZ, tabun (GA), sarin (GB), soman (GD), thickened
GD (TGD), sulfur mustard (HD), thickened HD (THD), lewisite (L), mustard-lewisite mixtures
(HL), V-agent (VX), and other hazardous materials and toxins, such as arsine (SA), cyanogen
chloride (CK), hydrogen cyanide (AC), phosgene (CG), perfluoroisobutylene (PFIB), as well as
agent simulants, Class A poisons, and toxins (e.g., T-2 toxin).
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The HMRC complex has approximately 10,000 sq ft of laboratory and support space. It includes
the Hazardous Materials Laboratory (HML) and the Large Item Test Facility (LITF), which
provide approximately 2,000 sq ft of laboratory space and 100 linear ft of CSM-approved filtered
hoods for working with neat (pure) CW agents; about 630 sq ft of Research Dilute Solution
(RDS, diluted chemical agent) laboratory space, including four fume hoods; approximately 2,100
sq ft of laboratory support areas, including wastewater and general laboratory waste disposal,
environmental monitoring, emergency power supplies, air filter systems, and general equipment
storage room; and about 800 sq ft of staff support areas, including personnel showers, change
rooms, laundry facilities, and other common use areas.
The MREF specializes in the research, development, test, and evaluation (RDT&E) of medical
countermeasures against highly pathogenic biological and highly toxic chemical materials. This
facility is one of a very limited number of U.S. laboratories capable of studying aerosolized
etiological agents in animal models under BSL-3 containment. The facility maintains state-of-
the-art equipment and professional and technical staffing expertise to safely conduct in vivo
testing and evaluation of hazardous biological materials under the FDA's GLP Guidelines (21
CFR Part 58).
The MREF facilities are ISO 9001 certified, accredited by the American Association for the
Accreditation of Laboratory Animal Care (AAALAC), and inspected and compliant with the
USDA, FDA, Drug Enforcement Agency, Ohio EPA, U.S. Army Safety Team, U.S. Army
Inspector General, U.S. Army Medical Research Institute of Chemical Defense Safety and
Chemical Operations Branch (SCOB), U.S. Army Medical Research and Materiel Command
Office of Animal Care and Use Review, Madison County Health Department, and Battelle's
Institutional Animal Care and Use Committee (IACUC). The MREF fully complies with all
applicable U.S. Army Regulations, and Federal Government and State of Ohio regulations to
conduct and support RDT&E studies using highly toxic chemical and pathogenic biological
materials. The MREF is licensed to ship, receive, and handle select agents, as defined by CDC.
The MREF BSL-3 facility was completed in 1995, and expanded in 2002 to consist of
approximately 31,000 sq. ft. The containment area within the facility is designed to meet or
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exceed the BSL-3 facility guidelines published by the CDC and NIH entitled Biosafety in
Microbiological and Biomedical Laboratories (4th edition, 1999). The seven BSL-3
microbiology laboratories contain multiple Class III biosafety cabinets (BSCs) linked together in
an H-shaped configuration and two autoclaves. Additional laboratories within this area include a
microbiology laboratory equipped with a Class II BSC connected to a Class III BSC, and a dose
configuration room equipped with a Class II BSC.
3.2 Site Operations
Battelle operates its certified chemical and biological test facilities in compliance with all
applicable Federal, state, and local laws and regulations, including U.S. Army regulations.
Battelle's facilities are certified through inspection by personnel from the appropriate
government agency. The HMRC has been certified to work with chemical surety material
through a Bailment Agreement by the U.S. Army Soldier Biological and Chemical Command
(SBCCOM). SBCCOM will terminate its Bailment Agreements on 1 September 2003, and
Battelle has begun the process to transition to an AR50-6 surety facility. In this transition,
Battelle will demonstrate, via inspections by the appropriate government personnel, that its
facilities meet all Federal, state, and local laws and regulations, including U.S. Army regulations.
Battelle operates the MREF in compliance with requirements contained in 32 CFR 626 and 627,
Biological Defense Programs. Our chemical and biological facilities and attendant certifications
are listed in Table 3-1.
Test procedures at the HMRC and MREF are governed by established standard operating
procedures (SOPs). Those documents are specified by facility, number, and title. In all cases,
the latest version of every such document is used. All relevant documents will be reviewed as
part of the Operational Readiness Inspection (ORI) for verification testing to identify whether
any test-specific modifications need to be implemented. The documents that are relevant to
testing are indicated where appropriate throughout this test/QA plan.
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Table 3-1. Certifications of HMRC and MREF
Facility
Materials
Level
Certification
Hazardous Materials
Research Center
Chemical warfare
agents
Chemical Surety
Materiel (CSM)
(Neat)
RDT&E (Dilute)
Bailment Agreement
No. DAAD13-H-03-0003
Analytical Chemistry
Laboratory
Chemical warfare
agents
RDT&E (Dilute)
Bailment Agreement
No. DAAD13-H-03-0003
Medical Research and
Evaluation Facility
Biological warfare
agents
Biosafety Level 3
CDC Select Agents Program
(32 CFR 626 and 627)
administered through the
Biological Defense Research
Program
Chemical warfare
Chemical Surety
United States of America
agents
Materiel (CSM)
(Neat)
RDT&E (Dilute)
Medical Research Materiel
Command (USAMRMC)
No. G472501
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4.0 EXPERIMENTAL DESIGN
4.1 General Test Design
This test/QA plan specifies procedures for testing hydrogen peroxide vapor decontamination
technologies with chemical and biological agents and surrogates at the laboratory scale using
small samples of indoor materials (i.e., coupons). Verification testing will determine efficacy of
the technology against agents with representative indoor surface materials. The verification test
design will also produce data that will allow correlations to be made between results with actual
agents and those with selected surrogates. In all testing, each decontamination technology will
be applied in a manner consistent with the manufacturer's recommendations. The technology
vendor will provide the equipment for application of their technology, and will train Battelle
staff in its use. The effect of the decontamination technology on indoor materials will also be
assessed by visual inspection of test coupons after they are subjected to decontamination.
The following subsections introduce the primary features of the verification testing approach.
Details on the procedures used to conduct testing are presented in Section 6.
4.1.1 Parameters to Be Tested
The following performance parameters of hydrogen peroxide vapor decontamination
technologies will be tested, using coupons of representative indoor materials contaminated with
biological and chemical agents in controlled laboratory tests:
• Efficacy in destroying chemical agents and surrogates on selected indoor surfaces
• Efficacy in destroying biological agents and surrogates on the same indoor surfaces
• Generation of toxic degradation products from interaction of the decontaminant with
the target chemical agents.
Qualitative biological indicators will also be used in testing, to allow correlation of their results
with the quantitative efficacy results. Information on surface damage caused by the
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decontamination technologies will also be gathered by visual inspection of the test coupons after
decontamination.
4.1.2 Scale of Testing
The performance parameters listed above will be evaluated through testing with chemical and
biological agents and surrogates at the laboratory scale. These performance tests will use small
coupons (approximately 3/4 in. x 3 in. (1.9 x 7.5 cm)) of selected indoor materials as test surfaces,
and will be carried out in a suitable chemical or biological agent safety hood or cabinet. Multiple
coupons of each of several indoor materials will be contaminated with the target agents, and then
treated with the decontamination technology. Blank (i.e., uncontaminated) and control (i.e.,
contaminated but not decontaminated) coupons will also be used for each test material, and will
serve as the basis for calculations of decontamination efficacy. This scale of testing will provide
a controlled, reproducible approach to assess efficacy with real agents, while also requiring a
realistic though small-scale application of the decontaminant.
4.1.3 Efficacy
Efficacy (the effectiveness with which the hydrogen peroxide vapor decontamination technology
destroys the agent) will be determined for both chemical and biological agents and surrogates by
means of the coupon tests. Efficacy testing will rely on comparing the amount of contaminant
on test coupons before decontamination (control coupons) to the amount present after application
of the decontamination technology (test coupons). Multiple coupons will be used for both the
control and test samples, and the resulting data will be used to calculate efficacy as a percent
removal (for chemical agents) or a log reduction (for biological agents).
For building decontamination, the residual amount of agents left after decontamination also
needs to be considered, since evaporation of, or physical contact with, any residual could carry a
health risk for building occupants. For chemical and biological agents, allowable residual levels
have not been determined.(4) However, a precedent has been set for the desired end result of
decontamination for biological agents: a 6-log kill of biological indicators, and additionally, no
growth is to be detected after decontamination. Though not an official level for building
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decontamination, biological efficacy testing under this test/QA plan will follow the 6-log kill
objective. For chemical agents, the allowable residual is undefined, but some exposure limits
have been set for vapor exposure and for contact hazards/4'5) Consequently, the post-
decontamination levels of chemical agent will be determined by three methods, including coupon
extraction, offgasing, and contact transfer. The extraction method will measure the percent
efficacy for destruction of chemical agent on the coupon surfaces. The offgasing method will
measure the amount of residual chemical agent vapors offgasing from a test material that could
create a vapor hazard. The contact transfer method will measure the amount of residual chemical
agent on the coupon surface that could potentially create a contact hazard when transferred to
skin or other material during contact.
Efficacy will be evaluated for each chemical and biological agent and surrogate, for each
selected indoor surface material. Biological efficacy testing will employ seven coupons of each
surface material: three contaminated and subjected to decontamination (test coupons), three
contaminated but not subjected to decontamination (control coupons), and one not contaminated
(blank coupon). A corresponding set of coupons will be used for chemical efficacy testing,
however, chemical testing will also employ additional coupons of each surface material, for the
vapor offgasing and contact transfer tests.
4.1.4 Temperature and Relative Humidity Conditions
Different hydrogen peroxide vapor decontamination technologies may require different humidity
conditions in the environment to be decontaminated, so that the technology can be most
effective. Coupon testing will be carried out at room temperature, and at whatever humidity
condition is required and/or maintained by the technology undergoing testing. Temperature and
humidity will be monitored during the decontamination process.
4.1.5 Surface Damage
The effect of decontamination on the indoor materials used as test surfaces will be evaluated
informally in conjunction with the efficacy testing procedure. After decontamination of the test
coupons, the appearance of the decontaminated coupons will be observed, and any obvious
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changes in the color, reflectivity, and apparent roughness of the coupon surfaces will be noted.
This comparison will be conducted for each of the test materials, before any extraction or
sampling of the decontaminated test coupons takes place.
4.2 Agents and Surrogates
The chemical and biological agents to be used in verification testing under this plan were
selected based on an evaluation of potential threats to buildings,(3) and on subsequent input from
stakeholder groups. Note that the threat summary was based on a survey of expert opinions, and
not on an exhaustive analysis. That evaluation considered the availability of potential
contaminants (including chemical and biological agents, biotoxins, and TICs), the lethality of the
contaminants, the potential delivery pathways for the contaminants, and the persistence of the
contaminants in a building. In addition to chemical and biological agents, surrogates will be
used in testing to establish correlations between the decontamination efficacy for surrogates and
actual agents.
4.2.1 Chemical Agents and Surrogates
The chemical agents to be used for verification testing, listed in order of priority, are:
Table 4-1. Chemical Agents to Be Used
Agent
Acronym
Type
Purity
V Series
VX
Nerve Agent
> 85%
H Series
HD
Vesicant
> 85%
Thickened Soman
TGD
Nerve Agent
AP
AP: As provided by the U.S Army; see text.
These agents are key representatives of families of similar agents. The agent specified in
thickened form (TGD) was chosen because the thickened matrix enhances the persistence of the
agent on surfaces. This agent will be obtained in thickened form from the U.S Army, and the
Army will provide information on the purity of the thickened agent. However, it will not be
possible to confirm the agent's purity by analysis, due to interference from the thickening agent.
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For each of the chemical agents listed above, a chemical surrogate will also be used. The
selection of chemical surrogates for testing of decontaminants is a complex issue. Possible
surrogates that have been identified include:
• Methyl parathion and Malathion for VX
• Methyl phenyl sulfide for HD
• Diisopropyl phosphonofluoridate for GD.
Previous use of these surrogates has been based on the similarity of their physical properties to
those of the chemical agents. Alternative choices of surrogates may be used, if evidence is found
that the alternative surrogates better mimic the chemical reactivity of the agents with hydrogen
peroxide vapor.
4.2.2 Biological Agents and Surrogates
The primary biological agent used in testing of the hydrogen peroxide vapor decontamination
technology will be anthrax spores (.Bacillus anthracis, Ames strain). To provide correlations
with the anthrax results, two biological surrogates will be used:
• Bacillus stearothermophilus (ATCC12980)
• Bacillus subtilis (ATCC 19659).
The B. stearothermophilus surrogate was chosen because previous tests have indicated it to
behave similarly to anthrax in response to gaseous decontaminants, and it has historically been
used as an indicator for hydrogen peroxide vapor because it is a particularly difficult organism to
kill using this technology. The B. subtilis (ATCC 19659) surrogate was chosen because it is the
same as used in the AO AC Sporicidal Activity Test. Anthrax and the two surrogate organisms
will be used in the form of spore suspensions, for application to the test surfaces.
A commercial spore strip will also be included in testing, of the same spore type (B. subtilis var
niger (B. atrophaeus (ATCC 9372)), backing (paper), and manufacturer (Raven) as those used
during decontamination of anthrax in U.S. Postal Service facilities. Furthermore, biological
indicators (Apex Labs) containing the surrogates B. stearothermophilus and B. subtilis will also
be included. These biological indicators will contain a spore population of 106. The Raven
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spore strips and Apex biological indicators will be used for qualitative indication of efficacy, to
allow correlation with quantitative efficacy results.
4.3 Test Surfaces
The surface materials to be used for testing hydrogen peroxide vapor decontamination
technologies are a subset of the variety of structural, decorative, and functional surfaces that may
be found indoors. Excluded from the list of test surfaces are indoor materials that are likely to be
removed from a contaminated building for disposal, rather than decontaminated in place. Such
materials include draperies, ceiling tiles, and fabric furnishings. However, the surface materials
to be used include both smooth and porous surfaces, and a variety of material compositions. The
test surfaces that will be used are listed below, with the unique code that will be used for sample
identification shown in parentheses:
• Painted (latex, semi-gloss) concrete (cinder block) (PC)
• Painted (latex, flat) wallboard (PW)
• Decorative laminate (DL)
• Galvanized ductwork (GM)
• Glass (GS)
• Bare wood (pine lumber) (BWD)
• Industrial grade carpet (IC).
The test coupons of each surface material will be 1.9 cm x 7.5 cm, with thickness varying from
1/32 to 3/8 as appropriate for the materials. Certain combinations of contaminant and test
surface have been avoided in making this selection. For example, hydrolysis of VX has been
shown to occur rapidly (half-life = 3 hours) on bare concrete surfaces.® Consequently, bare
concrete was avoided for testing of decontamination efficacy with VX because the substrate
efficacy would confound the determination of the decontaminant efficacy.
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4.4 Test Sequence
Table 4-2 provides the sequence of testing to be carried out on each technology, listing the
names of the test procedures, the performance or operational parameters to be evaluated in each
procedure, and a summary of the samples or data comparisons resulting from each procedure.
The order of testing will be as shown in Table 4-2, i.e., biological efficacy testing with coupons,
followed by corresponding chemical efficacy testing, in each case followed by assessment of
surface damage.
Table 4-2. Sequence of Test Procedures in Verification Testing of Hydrogen Peroxide
Vapor Decontamination Technologies
Test Procedure
Parameters Evaluated
Data Produced
Biological efficacy test
Efficacy
Multiple samples, plus controls and blank, for
each test surface, for each biological agent and
surrogate. Also, multiple spore strip samples,
plus controls.
Damage to surfaces
Damage to test coupons
Visual observation of every test coupon in all
biological efficacy tests.
Chemical efficacy test
Efficacy
Multiple samples, plus controls and blank, for
each test surface, for each chemical agent and
surrogate.
Damage to surfaces
Damage to test coupons
Visual observation of every test coupon in all
chemical efficacy tests.
Test for known toxic by-
products
Analysis of coupon extractions
after chemical efficacy tests
Multiple samples, plus blank, for each test
coupon/agent combination
Vapor offgas test for
chemical agents"
Effectiveness at reducing vapor
offgasing
Multiple samples, plus controls and blank, for
each test surface, for each chemical agent and
surrogate.
Contact transfer test for
chemical agents3
Effectiveness at reducing
contact transfer
Multiple samples, plus controls and blank, for
each test surface, for each chemical agent and
surrogate.
a: These tests will use separate coupons from those used for other test procedures.
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5.0 EQUIPMENT AND MATERIALS
This section provides a description of the key materials and equipment needed to perform
verification testing of hydrogen peroxide vapor technology.
5.1 Materials
5.1.1 Agents
Chemical agent use at the HMRC will be under the terms and conditions of Bailment Agreement
DAAD13-H-03-0003. This Bailment Agreement is a contract between Battelle and the U.S.
Army that specifies the safety, security, and personnel reliability standards required for work
involving the storage, handling, and use of chemical agents. Battelle's stock of agent will be
analyzed prior to testing to verify the purity of the agent used to contaminate the test coupons.
An aliquot of diluted agent will be injected into a gas chromatograph (GC) fitted with a flame-
ionization detector to determine the purity of the agent. The purity of the agent will be
determined through comparison with the analytical standards generated from or based on
Chemical Agent Standard Analytical Reference Material (CASARM). Only chemical agents
with purity greater than 85 percent will be used in this program. The purity of thickened GD will
not be measured due to interference caused by the thickener, but information on the agent purity
will be provided by the U.S. Army.
Biological agent use at the MREF will be according to the CDC Select Agents Program (42 CFR
Part 73) and the Biological Defense Research Program (32 CFR 626 and 627) in adherence with
the Battelle MREF Facility Safety Plan. Anthrax (Ames) spores will be prepared according to
MREF. X-074 (Production of Bacillus Anthracis Spores) or MREF. X-093 {Production of
Bacillus anthracis Spores in a Small Fermentor). The spores will be characterized according to
MREF. X-075-00 (Characterization and Qualification of Bacillus anthracis Spores), which
requires less than 5 % debris content for acceptance of spores.
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5.1.2 Spore Strips
The Raven commercial spore strips and Apex Labs biological indicators will be purchased for
verification testing, in quantities larger than needed for testing.
5.1.3 Surfaces to Be Tested
Section 4.3 lists the materials to be used to simulate indoor surfaces in testing. The
representativeness and uniformity of the test materials are important to assure reliable test
results. Representativeness means that the materials used are typical of such materials used
indoors in buildings. Uniformity means that all test pieces are essentially equivalent for the
purposes of testing. Representativeness will be assured by obtaining test materials from
appropriate suppliers and by recording the appropriate specifications, manufacturer
identification, lot numbers, etc., for each material.. Uniformity will be maintained by obtaining a
large enough quantity of material that multiple test samples of uniform characteristics can be
obtained (e.g., test coupons will all be cut from the interior rather than the edge of a large piece
of material). In addition, the uniformity of recovery of biological and chemical agents will be
assessed for each test material, in method demonstration tests conducted before the start of
verification testing (see Section 6.1). The reproducibility of recovery rates will be determined
for each material, as a measure of the uniformity of the test pieces.
5.2 Delivery and Application Equipment
5.2.1 Agent/Surrogate Surface Application
The equipment needed to apply controlled and reproducible amounts of agents and surrogates to
the test surfaces will include the solutions or suspensions to be delivered, and the delivery device
(a syringe, pipette, or comparable system). These items of equipment are described in Section 6.
5.2.2 Temperature/Humidity Conditions
Commercial decontamination technologies based on hydrogen peroxide vapor typically include a
conditioning system that controls the temperature and/or humidity of the environment to the
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optimum conditions for the decontamination. In all verification testing, each technology will be
operated according to the vendor's instructions, including the performance of any such
conditioning system. The temperature and humidity of the test enclosure will be monitored
throughout testing, using vendor provided sensors.
5.3 Test Chamber
A decontaminant exposure chamber will be used to expose the test coupons to the
decontaminant. For biological agent testing, a Compact Glove Box Model 830-ABC (Plas Labs,
Inc., Lansing, MI; Figure 5-1) will be used. This unit has inner dimensions of 28"w x 23"d x
29"h (71cm x 59cm x 74cm) and outer dimensions of 43"w x 24"d x 31"h (110cm x 61cm x
79cm). The unit also has a top opening of 17" x 23" (43cm x 58cm) and a transfer chamber that
is 12" (30cm) long and an inner diameter of 11" (28cm). The chamber has a total volume of
11.2cubic feet (317 L). A set of glove ports, located on the side, are available for working in the
hood. The same type of glove box, but without the transfer chamber, will be used in the
chemical agent testing. In both cases, the decontaminant will be directed from the vendor's
delivery system through the exposure chamber, at the temperature and humidity conditions
established by the delivery system.
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Figure 5-1. Compact Glove Box—BW Agent Tests
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5.4 Sampling and Analysis Materials and Equipment
5.4.1 Chemical Agent Testing
5.4.1.1 Contact Transfer Equipment. The contact transfer test equipment will include 2-inch
diameter pieces of latex dental dam for contact transfer measurements, and 1-inch diameter
weights (65 g/cm ) for placing on the latex. Figure 5-2 shows the contact transfer test with the
weights applied to the test coupons. The latex will be washed with water and dried at 185°F for
24 hours prior to cutting.
Figure 5-2. Contact Transfer Weights
5.4.1.2 Offgas Sampling Equipment. Offgas sampling will be performed at ambient laboratory
conditions of temperature and relative humidity. The offgas rack (Figure 5-3) will hold up to
25 test cells. Aluminum offgas cells (Figure 5-4) will be used to hold the individual coupons
during offgasing, and may include critical orifices to control the flowrate at 0.25 L/min during
offgas collection. Butyl o-rings will be used to seal the cells. Charcoal tubes will be placed on
the cell inlet to ensure clean air is entering the cell. The agent vapors will be collected using
solvent-filled impingers or sorbent tubes, depending on the agent and amount of agent offgasing
expected from each material type. The agents will be collected in bubblers filled with diethyl
phthalate or ethylene glycol diacetate, or with sorbent tubes filled with either Carboxen or Tenax.
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Figure 5-3. Offgas Rack
Figure 5-4. Offgas Cell
The Carboxen tubes will be solvent extracted for analysis. The Tenax sorbent tubes will be
thermally desorbed to analyze for the chemical agents.
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5.4.1.3 Analytical Equipment. Chemical agent analyses will be performed using Hewlett
Packard 5890 or 6890 GCs equipped with flame photometric detectors (FPD) or flame ionization
detectors (FID). Internal standard (IS) will be added to the chloroform to produce a
concentration of 1.7 jig/mL using triethyl phosphate (TEP) and diethylmethylthio-
methylphosphate (DEMTMP). The TEP is used as the IS for GD analysis, and DEMTMP is
used as the IS for HD and VX analysis. The internal standard for the surrogate analysis will be
either TEP or DEMTMP, based on the relative retention times of the surrogates and the IS.
5.4.2 Biological Agent Testing
5.4.2.1 Sampling Media. The procedures and media used to extract the biological agent or
surrogates from the test surfaces are described in Section 6.2.5.2.
5.4.2.2 Sample Analysis. Section 6.2.5.2 describes the culturing and enumerating procedures for
biological samples.
5.5 Performance Evaluation Audit Materials
The performance evaluation (PE) audit (Section 7.2) will use independent standards to check the
analysis methods for chemical agents and chemical surrogates. These independent standards will
be RDS solutions, prepared in Battelle's Columbus facilities, and analyzed with the GC
equipment used for sample analysis at the West Jefferson facilities. At least one such RDS
solution will be prepared for each of the chemical agents and surrogates identified in Section
4.2.1.
No PE audit will be done for biological agents, due to the lack of suitable audit standards. The
application confirmation procedure (Section 6.2.3.2), controls, blanks, and method validation
procedures will be used to document the biological test results.
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6.0 TEST PROCEDURES
This section provides a discussion of procedures for method validation, and for chemical and
biological coupon testing of hydrogen peroxide vapor decontamination technologies.
6.1 Method Demonstration
Many of the test coupon materials to be used are likely to be new to decontamination testing.
Consequently, method trials will be performed as necessary to demonstrate the methods included
in this test/QA plan.
6.1.1 Chemical Agent Method Demonstration
Method demonstration will be performed as necessary to determine the optimum methods for
extraction of chemical agents and surrogates from the various test coupons, and for quenching
the hydrogen peroxide vapor reactions to analyze for chemical agent, surrogates, and toxic
degradation products after decontamination. Demonstration trials will be conducted with each
chemical agent/surrogate/coupon combination, using 5 percent bleach as the positive control, and
distilled water as the negative control. Extraction efficiencies for various solvents (e.g.,
chloroform, hexane) from these combinations will be determined. Furthermore, prior to testing
one coupon of each material type, and the samplers used as the contact transfer material (see
Section 6.2.5.1.3) will be extracted in solvent to ensure that there are no analytical interferences
that would inhibit agent analysis. Up to three solvents will be tested.
The objective of the quenching demonstration study is to establish a quenching method that will
stop the reaction between the hydrogen peroxide vapor and the chemical agent. Typically
samples are quenched with an organic compound containing sulfur that is also soluble in the
extraction solvent to stop oxidation of the agent. Sodium sulfite has often been used for this
purpose, as it reacts very rapidly with hydrogen peroxide vapor. In addition, dilution or
extraction may also be found effective at quenching the reaction. Up to four methods will be
tested. Based on the results of method trials, an appropriate extraction solvent will be selected.
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The solvent selection process will consider extraction efficiencies, analytical interferences,
material compatibility, and other observations made during the trials.
In addition, trials of the offgasing method will be performed to determine the appropriate vapor
collection system design based on the amount of agent vapor leaving the various coupons. Trials
will be performed on each type of test coupon using each type of agent. Results will be used to
determine how many coupons should be in the offgasing cell, and what is most the most
appropriate system for collection of vapors (e.g., solvent fill impingers, sorbent filled tubes, or
both, which solvent, which sorbent, etc.).
6.1.2 Biological Agent Method Demonstration
Method demonstration trials will be performed as necessary to determine the optimum methods
for extraction of biological agents from the test coupons and for quenching the decontamination
reaction. The objective of the quenching demonstration study is to determine a quenching
method that will stop the reaction between the decontaminant and the biological agent so that
decontamination does not continue after sampling. For example, a solution of sodium
metabisulfite can be used to quench the reactivity of some decontaminants that act by oxidation.
Once a method has been established, the method demonstration trials will determine the average
spore recovery efficiency from each type of surface material, and the reproducibility of that
efficiency. The reproducibility will be determined as a percent coefficient of variation (%CV) of
repeated trials with each surface material. The %CV values will indicate the uniformity of the
test coupons for each material. The average recovery values will determine what log kill can be
determined based on an initial spore loading of 108.
6.2 Coupon-Scale Testing
Decontamination efficacy testing with coupons will be conducted based on procedures described
in TOP 8-2-061 {Decontamination Systems Laboratory/Field Testing). The testing will evaluate
decontamination efficacy for chemical and biological agents by extracting and measuring the
initial and residual agent on test coupons. Chemical analysis or biological enumeration of the
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resulting extracts will allow efficacy to be calculated as the percent removal for chemical agents,
or the log reduction for biological agents.
For chemical agents, as discussed in Section 4, measurements will also be made of vapor
offgasing and contact transfer, to help in the evaluation of efficacy from the perspective of
residual chemical agent allowed to remain in a building after decontamination. The offgasing
tests will measure the amount of chemical agent vapors evaporating from the coupon, potentially
creating a vapor hazard. Contact transfer tests will measure the amount of chemical agent
transferred to a simulated skin material touching the coupon surface, simulating a contact hazard
when transferred to skin or other material. Detailed descriptions of these tests are presented in
Section 6.2.5.1.
6.2.1 Preparation of Test Materials
For testing of chemical agent decontamination, no special preparation of test surfaces is required.
To ensure normal cleanliness and prevent contamination of test surfaces, care will be exercised
and the test coupons will be packaged in individual sample bags. At most, surface preparation
will involve washing with a solvent or water. The test coupons will be cut to 1.9 cm x 7.5 cm
size from the interior of a large piece of test material. Edges and damaged areas will be avoided
in cutting test coupons. The test coupons will be visually inspected upon receipt and any
evidence of damage will be recorded. The length, width, and thickness of the test coupons will
be measured and recorded.
Chain-of-Custody (CoC) forms will be used to ensure that the test coupons are traceable
throughout all phases of testing. Each coupon will be assigned a unique identifier code that
matches it with the sample, test parameters, and sampling scheme. The testing staff receiving the
test coupon will be responsible for comparing the identifier code with the test matrix.
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6.2.2 Application of Agents to Test Coupons
6.2.2.1 Application of Chemical Agents and Surrogates to Test Coupons
To assess decontamination efficacy, the conditions specified in TOP 8-2-061 will be used, i.e.,
a contamination density of 10 g/m2, and a droplet size of 1-jo.L. For the three chemical agents,
the number of agent drops to be administered per coupon will be determined prior to testing
based on the contamination density, agent density, and the measured agent purity.
The test coupons will be removed from their individual packages and allowed to equilibrate to
the laboratory temperature and relative humidity for a minimum of one hour prior to agent
application. A 1-inch diameter circle will be drawn on the test coupons with a non-interfering
grease pencil, to provide a known area for agent application.
The test coupons will be placed so they are lying flat in the chemical agent fume hood. The
chemical agents will be applied to the test coupons using microliter-sized drops to achieve the
target contamination density (10 g/m2). A Hamilton gastight syringe with a Hamilton repeatable
stepper will be used to produce the drops. Separate syringes will be used for each chemical
agent to prevent cross-contamination. After agent application, the coupons will be covered with
a Petri dish to minimize agent evaporation. Coupons will be allowed to weather overnight (i.e.,
approximately 16 to 18 hours) after application of chemical agent. SOP HMRC-II-001 {General
Provisions for Handling Chemical Agent in the Hazardous Materials Research Center) will be
used for agent operations. The same procedures used for application of the chemical agents will
also be used for application of the surrogates.
6.2.2.2 Application of Biological Agents and Surrogates to Test Coupons
Testing will be performed in a Compact Glove Box (Plas Labs, Inc.) (see Section 5.3). Test
coupons will be placed lying flat in the cabinet, and contaminated at challenge levels of 108
spores per coupon. Stock suspensions of the agent at the required concentration will be
prepared, transferred to the coupon using a micropipette, and spread over the sample surface
(e.g., by smearing the suspension over the coupon with the tip of the pipette, or placing the
suspension over the surface as small droplets similar to the chemical agent approach). After
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contamination with biological agent or surrogate suspension, the test coupons will be allowed to
dry undisturbed to completion.
6.2.3 Confirmation of Surface Applications
6.2.3.1 Confirmation of Surface Application Density of Chemical Agents
Each chemical agent will be applied to three Teflon control coupons at the desired density using
the procedure described in Section 6.2.2.1. These coupons will be extracted using the same
procedure used for the decontaminated coupons (see Section 6.2.5.1.1) immediately after agent
application. They will be analyzed by the same procedure used for decontaminated coupons (see
Section 6.2.5.1.4), to verify the initial application density.
6.2.3.2 Confirmation of Surface Application Density of Biological Agents
To confirm the application density of biological agents and surrogates, the anthrax and surrogate
spore suspensions used to contaminate the coupons will be re-enumerated on each day of use.
This enumeration will be carried out as described in Section 6.2.5.2.
6.2.4 Application of Hydrogen Peroxide Vapor Decontamination Technology
After application of agents and surrogates to the test coupons, and completion of the drying or
weathering time, the test coupons will be decontaminated. Each decontamination technology
undergoing testing will be used in accordance with the vendor's instructions, to supply the test
enclosure with the required levels of hydrogen peroxide vapor for decontamination. The
duration and hydrogen peroxide vapor level used for decontamination will be as recommended
by the vendor.
If feasible, spectroscopic monitoring of hydrogen peroxide vapor will be conducted during the
decontamination in accordance with vendor's instructions.
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6.2.5 Determination of Decontamination Efficacy
The primary test of decontamination efficacy will determine the fraction of agent destroyed by
the hydrogen peroxide vapor treatment, through extraction of residual agent from the coupons
after decontamination. In addition, analysis of end point conditions will be made (i.e., by
performing vapor offgasing and contact transfer testing for chemical agents, and verifying no
growth for biological agents). The vapor offgasing and contact transfer tests will provide
alternative measures of efficacy for the chemical agents and surrogates.
6.2.5.1 Decontamination Efficacy for Chemical Agent on Coupons
6.2.5.1.1 Extraction of Residual Chemical Agent from Coupons
After application of the decontaminant, extraction of the residual chemical agent will be
performed. Decontaminated test coupons and the control coupons will be placed directly into
jars containing the extraction solvent. After a 1-hour extraction, an aliquot of the solvent will be
transferred to a GC vial for analysis. Depending on the outcome of the method validation effort,
a phase separation may be performed to minimize analytical interferences, by separation of
coupon debris from the extraction solution. The sample will be analyzed for chemical agent
using a GC with an appropriate detector as discussed in Section 6.2.5.1.4.
For chemical agents, decontamination efficacy will be calculated based on the amount of agent
applied to the test coupon and the amount of residual agent measured after decontamination, as
described in Section 8.2.2. Decontamination efficacy results will be presented as percent agent
neutralized/removed. The upper limit for calculated efficacy values is based on the detection
limit of the GC and the amount of solvent used for extraction; typically these limitations do not
come into play unless efficacy exceeds 99.9%.
6.2.5.1.2 Offgasing Measurements
The offgasing test will be performed using different coupons than are used for extracting the
residual agent for the primary determination of efficacy (Section 6.2.5.1.1). Larger coupons may
be required for the offgasing test in order to produce sufficient agent vapor for analysis.
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The vapor offgasing test will be performed on coupons that have been contaminated with
chemical agent and subsequently decontaminated with the hydrogen peroxide vapor, as well as
on contaminated coupons that have not been decontaminated (i.e., control coupons). Each
coupon will be sealed in an aluminum offgas cell. A charcoal filter will be placed on the cell
inlet to provide clean airflow into the cell. A sorbent tube or impinger will be attached to the cell
exhaust. Critical orifices or mass-flow controllers will be used to control the flow through the
sorbent tubes or impingers at 0.25 liters per minute. The offgas will be sampled over specific
time intervals of 0-2, 2-4, and 4-12 hours. The sorbent tubes will be extracted with 3 mL of
solvent and analyzed for chemical agent by GC. The impinger solutions will analyzed directly
by GC, or extracted prior to analysis by GC. SOP HMRC-X-049 (Offgas Testing of Materials)
will be followed for this test. The efficacy of reducing the vapor offgasing will be calculated by
comparing the offgasing rates for the decontaminated coupons to those from the control coupons,
as described in Section 8.2.4.
6.2.5.1.3 Contact Transfer
The contact transfer test will be performed using different coupons than used for the vapor
offgasing test (Section 6.2.5.1.2) or for extracting the residual agent for the primary
determination of efficacy (Section 6.2.5.1.1). The contact transfer test will be performed after
the vapor offgasing test, for both the decontaminated and the control coupons.
The amount of agent transferred by contact will be measured using a piece of latex dental dam
(dental dam is made from natural rubber latex and other ingredients and is used as a barrier
during endodontic and other restorative procedures). A 2-inch diameter piece of latex will be
placed on the test coupon as a sampler. A 2-inch piece of aluminum foil will be placed on top of
the latex, and a 2-inch weight (65 g/cm2) will be applied to simulate the force of a hand touching
the surface. After 15 minutes of contact, the weight will be removed and the latex sample will be
placed in a jar containing 20 mL of solvent. After a 60-minute extraction, an aliquot of the
solvent will be transferred to a GC vial for GC-FPD analysis. If the agent concentration is below
the GC-FPD detection limit, a 10-mL aliquot of the solvent extract will be evaporatively
concentrated to 1 mL and reanalyzed. If the agent concentration is still below the GC-FPD
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detection limit, it will be reported as a non-detect. Contact Transfer and Offgas Testing
Following Chemical Agent Contamination and Decontamination (SOP HMRC-X-070) will be
followed for this test. The efficacy of reducing the contact transfer of agent will be calculated
by comparing the offgasing rates for the decontaminated coupons to those from the control
coupons, as described in Section 8.2.3.
6.2.5.1.4 Sample Analysis
A quantitative analysis of chemical agent in coupon extracts, latex extracts, and vapor offgas
samples will be conducted using GC coupled with flame ionization (FID), flame photometry
(FPD), or mass spectrometry (MS) detectors. Analysis will be performed and standards for
reference analysis will be prepared in solvent using neat agent in accordance with HMRC
Standard Operating Procedures IV-056-06 (Standard Operating Procedure for Operation and
Maintenance of Gas Chromatographs and for the Analysis of Solutions Containing GA, GB, GD,
GF, HD, VX by Gas Chromatography. Analytical standards will be generated from or based on
the CASARM standard (see Section 5.1).
The selection of a detector is based on the chemical agent being analyzed, the expected
concentration range, any interferences identified during the method validation process, and the
time required for analysis. The FPD will be used for the chemical agents and surrogates because
it has the highest sensitivity for measurement, and can also be used to analyze more samples per
day. The FID will be used if the agent concentration in the samples is high.
Extracts from the test coupons will be analyzed for specific degradation products. A possible
degradation product of VX, EA 2192, will be determined by liquid chromatography/mass
spectrometry (LC/MS/MS). SOP HMRC-III-001 (General Provisions for RDTE Dilute Solutions
Utilized in JN-4) will be used for handling laboratory samples.
The analytical results for each extract will be fitted to the calibration curve for the specific GC
used to analyze the extract. The agent concentrations for each extract will be determined by
Equation 1:
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(1)
where
C = agent concentration (|j,g/mL)
M = slope of the calibration line
A = the peak area (PA) for the agent, normalized to the internal standard peak area
w = Y intercept of the calibration line.
As given in Equation 1, the agent concentration for each sample is determined from the ratio of
the IS concentration to that of the agent. Analytical results in excess of the daily MDL for the
instrument will be recorded in p,g/mL. The agent (density) on the coupons will be determined by
Equation 2.
6.2.5.2 Decontamination Efficacy for Biological Agent on Coupons
6.2.5.2.1 Extraction of Spores from Test Coupons
The testing will quantify decontamination efficacy by measuring the anthrax or surrogate spores
on surface material coupons, both those exposed (test coupons) and unexposed (control coupons)
to the decontaminant. Following the decontamination process, each surface material coupon will
be placed in a 50 mL test tube containing 10 mL of sterile PBS with 0.1% Triton X-100 and
catalase. The purpose of the Triton X-100 is to minimize clumping of spores, and the purpose of
the catalase is to neutralize any residual hydrogen peroxide. For spore extraction, the tubes will
be agitated on an orbital shaker for 15 min at room temperature. Samples will then be heat-
shocked at 60 °C for 1 hr to kill any vegetative bacteria. Following heat-shock, 1.0 mL of each
extract will be removed, and a series of dilutions through 10"7 will be prepared in sterile water.
(2)
where
R = residual agent density (|j.g/cm2)
C = GC concentration (jag/mL) from Equation 1
Ev = extract volume (mL)
Sa = contaminated surface area (cm2).
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An additional qualitative assessment of hydrogen peroxide vapor efficacy will be conducted
following spore extraction. After the extraction process described above, each coupon will be
transferred to a clean 50 mL tube containing 20 mL of liquid nutrient broth. The vials will be
sealed and incubated overnight at 37°C on an orbital shaker. The next day, the tubes will be
assessed qualitatively for viability as "growth" or "no growth."
6.2.5.2.2 Enumeration of Spore Samples
The number of viable spores present on the surface materials will be determined using the
coupon extracts produced by the procedure in Section 6.2.5.2.1. Spore viability will be
determined by dilution plating, using both the undiluted extracts, and the successive dilutions of
each extract, to assure that accurate spore counts are achieved. One hundred microliters of the
undiluted extract and of each serial dilution will be plated onto Tryptic Soy Agar (TSA) plates in
triplicate, allowed to dry, and incubated overnight at 35 to 37 °C for B. anthracis and B. subtilis
and at 55 to 60 °C for B. stearothermophilus. Plates will be enumerated the next day, and the
colony forming units (CFU)/mL will be determined by multiplying the average number
of colonies per plate by the reciprocal of the dilution, as described in MREF SOP X-054
(.Enumeration of BL-2 and BL-3 Bacterial Samples Via the Spread Plate Technique). Data will
be expressed as mean + standard deviation (SD) of the numbers of colony forming units
observed. To calculate the efficacy of the decontamination treatment, the number of spores
remaining on the decontaminated test coupons will be compared to the number of spores on the
control coupons. Efficacy for biological agents will be calculated in terms of a log reduction, as
described in Section 8.2.2.
6.2.5.2.3 Qualitative Indicators
The spore strips and biological indicators will be exposed to the decontamination treatment along
with the surface material coupons, but will be used to determine only qualitative (i.e., growth/no
growth) efficacy, as described in Section 6.2.5.2.1 for the material coupons. Following the
decontamination process, the spore strips and biological indicators will be placed in liquid
nutrient broth, and the presence of any viable spores will be determined. No enumeration of the
spores will be attempted.
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6.2.6 Observation of Surface Damage
Following application of the decontamination technology, each test surface will be examined
visually to establish whether use of the decontamination approach caused any obvious damage to
the surface. Observation of surface damage will be done immediately after completion of the
decontamination process but before post-decontamination sampling to assess efficacy. If wetted
by the decontamination process, the test surface will be allowed to dry before any inspection for
damage. Visual inspection of the surface will then take place through side-by-side comparison
of the decontaminated test surface and the control coupons of the same test material. Differences
in color, reflectivity, contrast, and roughness will be assessed in this way. These observations
will be made by the testing staff and recorded.
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7.0 QUALITY ASSURANCE/QUALITY CONTROL
7.1 Equipment Calibrations
The methods to be used for the determination of chemical and biological agents and surrogates
are described in Section 6. The analytical equipment needed for these methods will be
maintained and operated according to the quality requirements and documentation of the test
facility. All equipment calibrations will be conducted with appropriate standards on a pre-set
schedule, and calibration results will be clearly and consistently recorded.
Hewlett Packard GCs will be used for analysis of the extract, offgas, contact transfer, and
residual rinse samples. For GC analysis, five calibration standards will be analyzed at the
beginning of each sample analysis. The GC will be recalibrated if the correlation coefficient (R2)
from the regression analysis of these standards is less than 0.99. In addition, the percent bias for
the low standard must be less than 25%, and the percent bias for the remaining standards must be
less than 15%. One or two calibration check standards will be run for every five samples. The
criteria for evaluation of the GC performance is listed below:
• R should be greater than 0.99
• The % bias for the lowest standard should be less than 25%
• The % bias for the remaining standards should be less than 15%
• For duplicate samples, the difference between should be less than 20%
• The areas of the internal standards should be within 40% of the average of the
standards
• The difference between the shortest retention time and the longest retention time for
the target agent should be less than 0.5 minutes.
The calibration range and associated example detection limits for each agent are listed in
Table 7-1. In the event that agent concentrations are above the highest calibration level for the
GC-FPD analysis, the samples will be analyzed using a GC equipped with an FID, or diluted and
reanalyzed by GC-FPD. The GC-FID will be calibrated in a range of 10 to 250 jig/mL.
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Table 7-1. Calibration Ranges
Extraction
Offgas
Contact
Calibration Range
((ig/cm2)
(|ig/cm2)
(jig/cm2)
TGD
0.1 to 10 |xg/mL
0.5 to 50
0.03 to 3
0.1 to 10
HD
0.5 to 10 |j.g/mL
2.5 to 50
0.15 to 3
0.5 to 10
VX
0.25 to 10 (xg/mL
1.25 to 50
N/A
0.25 to 10
* Actual detection limits depend on contamination area and extract volume.
The critical orifices and mass-flow controllers used for flow control in the offgas test will be
calibrated using a Buck Calilogger. The flowrate of 0.25 L/min will be used for the offgas
testing.
7.2 Assessment and Audits
7.2.1 Technical Systems Audits
Battelle's Quality Manager will perform a technical systems audit (TSA) once during the
performance of this verification test. The purpose of a TSA is to ensure that verification testing
is being performed in accordance with the test/QA plan and that all QA/QC procedures are being
implemented. In this audit, the Quality Manager may review the sampling and analysis methods
used, compare actual test procedures to those specified in this test/QA plan, and review data
acquisition and handling procedures. The Quality Manager will prepare a TSA report, the
findings of which must be addressed either by modifications of test procedures or by
documentation in the test records and verification report.
At EPA's discretion, EPA QA staff may also conduct an independent on-site TSA during
verification testing. The EPA TSA findings will be communicated to testing staff at the time of
the audit, and documented in a TSA report.
7.2.2 Performance Evaluation Audit
A PE audit will be conducted to assess the quality of the chemical agent and surrogate analyses
made during verification testing. This audit addresses only those measurements that factor
directly into the data used for verification, i.e., the decontamination technology is not the subject
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of the performance evaluation audit. Similarly, auxiliary measurement systems used to establish
test conditions (e.g., temperature, RH, and flow measurement devices) are subject to their own
usual calibration requirements, but are not subject to the PE audit.
The PE audit of chemical measurements will be made by independently preparing RDS solutions
of the agents and surrogates, in the same solvent and with the same nominal concentrations as
the calibration solutions used for the GC analysis. Successive analysis of these independent
solutions will then be conducted as a check on the calibration solutions. An acceptable tolerance
of ±25% will apply to this comparison. Failure to meet this criterion will require re-preparation
of the independent test solutions; a subsequent failure will trigger investigation of the calibration
process and flagging of test data for the agent or surrogate. This audit will be the responsibility
of Battelle, and will be carried out once during verification testing. Battelle's Quality Manager
will assess PE audit results.
No PE audit will be done for biological agents and surrogates, as quantitative standards for these
materials do not exist. The confirmation procedure, controls, blanks, and method validation
efforts will be the basis of support for biological test results.
7.2.3 Data Quality Audit
Battelle's Quality Manager will audit at least 10 percent of the verification data acquired during
verification testing. The Quality Manager will trace the data from initial acquisition, through
reduction and statistical comparisons, and to final reporting. All calculations performed on the
data undergoing audit will be checked.
7.2.4 Assessment Reports
Each assessment and audit will be documented in accordance with the QMP for the ETV
Building Decontamination Technology Center.(1) Assessment reports will include the following:
• Identification of any adverse findings or potential problems
• Space for response to adverse findings or potential problems
• Possible recommendations for resolving problems
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• Citation of any noteworthy practices that may be of use to others
• Confirmation that solutions have been implemented and are effective.
7.2.5 Corrective Action
The Quality Manager during the course of any assessment or audit will identify to the technical
staff performing experimental activities any immediate corrective action that should be taken.
If serious quality problems exist, the Quality Manager is authorized to stop work. Once the
assessment report has been prepared, the Verification Test Coordinator will ensure that a
response is provided for each adverse finding or potential problem, and will implement any
necessary follow-up corrective action. The Quality Manager will ensure that follow-up
corrective action has been taken.
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8.0 DATA ANALYSIS AND REPORTING
8.1 Data Acquisition
Data acquisition during verification testing includes proper recording of the procedures used in
testing, to assure consistency in testing and adherence to this test/QA plan; documentation of
sampling conditions and analytical results for the reference methods; determination of damage to
surfaces from the decontamination process; and recording of efficacy results and test conditions.
These forms of data acquisition will be carried out by the Battelle testing staff, in the form of test
notebooks, analytical data records, and data recording forms. Appendix A shows examples of
Test Performance Control Sheets (TPCSs) and a Test Coupon Sample Form that will be used
during testing.
Laboratory analytical data (e.g., method results quantifying the chemical or biological
contaminants used) may be produced electronically. Other test data will be recorded manually in
laboratory notebooks or on data forms prepared prior to the test. These records will be reviewed
to identify and resolve any inconsistencies. All written records must be in ink. Any corrections
to notebook entries, or changes in recorded data, must be made with a single line through the
original entry. The correction is then to be entered, initialed, and dated by the person making the
correction. A brief explanation of the basis for the correction will also be recorded.
Strict confidentiality of test data will be maintained. At no time will Battelle staff engage in any
comparison of the technology undergoing testing with any other decontamination technologies.
Table 8-1 summarizes the types of data to be recorded; how, how often, and by whom the
recording is made; and the disposition or subsequent processing of the data. The general
approach is to record all test information immediately and in a consistent format throughout all
tests. This process of data recording and compiling will be overseen by the Battelle Verification
Test Coordinator and Quality Manager.
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Table 8-1. Summary of Data Recording Process for Verification Testing
Data to Be
Recorded
Where
Recorded
How Often
Recorded
Disposition of
Data
Dates, times of test events
Laboratory record
books, data forms
Start/end of test, and at
each change of a test
parameter
Used to organize/ check test
results; manually incorporated
in data spreadsheets as
necessary
Test parameters
(agent/surrogate identities
and concentrations, test
surfaces, temperature and
relative humidity, gas flows,
etc.)
Laboratory record
books, data forms
When set or changed, or
as needed to document
the sequence of test.
Used to organize/ check test
results, manually incorporated
in data spreadsheets as
necessary
Sampling data
(identification of sampling
media, sampling flows, etc.)
Laboratory record
books, data forms
At least at start/end of
reference sample, and at
each change of a test
parameter
Used to organize/ check test
results; manually incorporated
in data spreadsheets as
necessary
Chemical analysis
or biological enumeration
analysis, chain of custody,
and results
Laboratory record
books, data sheets,
or data acquisition
system, as
appropriate.
Throughout sample
handling and analysis
process
Transferred to spreadsheets
Records and observations on
decon use
Laboratory record
books
Throughout
implementation of
decon technology;
during discussions with
decon vendor
Reviewed and summarized to
support data interpretation
8.2 Calculation Procedures
8.2.1 Data Screening
ANOVA models will be fitted to the residual extraction, contact transfer, and offgas results for
each agent (TGD, HD, and VX). Factors to be included in the models will be material, agent,
hydrogen peroxide vapor concentration, and time, as appropriate. Data will be checked for
normality and equal variance between groups and appropriate transformations (log) taken if
necessary. Outliers with normalized residuals greater than 3 standard deviations will be
considered for removal from the data.
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8.2.2 Efficacy
The primary assessment of efficacy will rely upon the comparison of the concentration of the
target agent or surrogate on the test coupons, before and after the application of the
decontamination technology. For chemical agents and surrogates, efficacy (E) in percent will be
calculated as
where C0 is the concentration of agent or surrogate before decontamination (determined from the
control coupons of each surface material) and Cf is the concentration on the test coupons after
decontamination.
For biological agents and surrogates, decontamination efficacy will be calculated as the log
reduction in viable organisms achieved by the decontamination technology. That is, efficacy (E)
for biological agents or surrogates will be calculated as
where N° is the number of viable organisms present on the control coupons (i.e., those not
subjected to decontamination), and N is the number of viable organisms present on the test
coupons after decontamination.
A separate efficacy calculation will be made for each of the surface materials, with each
chemical agent/biological agent/surrogate. In addition, since each surface material will be
represented by multiple sample coupons of that material in the efficacy tests, each combination
of a material and an agent/surrogate will result in multiple values of percent efficacy or log
reduction. For each material and agent/surrogate combination, a mean and range of the efficacy
values will be reported. Thus, the primary efficacy results from the coupon testing will be a
matrix table in which each entry shows the mean and range of efficacy results for one of the
agents/surrogates on one of the surface materials.
E = (C0 - Cf)/Co'100%
(3)
E = log (N°/N)
(4)
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8.2.3 Contact Transfer
The contact transfer of chemical agent is calculated based on the amount of agent transferred to
the sampler and the surface area sampled. Contact transfer (CT) is calculated according to
Equation 5:
CT=— (5)
A
where M is the mass of agent (in mg) collected on the latex contact surface of area A (in cm2).
The units of CT thus are mg/cm2.
The effectiveness with which the decontamination technology reduces the chemical contact
transfer will be calculated in a manner analogous to Equation 3, i.e.:
Ect = (CT0 - CTf)/CT0 • 100 (6)
where Ect is the percent efficacy for reducing contact transfer, and CT0 and CTf are the contact
transfer rates determined from the control and test coupons, respectively.
The residual contact hazard is estimated based on the contact transfer, the surface area contacted,
and the estimated hazard level for the percutaneous exposure to chemical agents. Criteria for
contact hazard estimation are defined in the NBC Contamination Survivability Criteria for
Military Equipment.(5) This document defines negligible risk percutaneous contact transfer
values for chemical agents, with negligible risk defined as mild incapacitation for 5 percent of
the military personnel. These values are listed below. Much lower levels would have to be
established for the general public as these numbers reflect battleground risks.
GD
VX
HD
30 mg/70-kg man
1.4 mg/70-kg man
180 mg/70-kg man.
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8.2.4 Offgas Flux
The offgas flux is calculated based on the amount of agent transferred to the sampler and the
surface area sampled. Offgas flux (OF) is calculated according to Equation 7:
where M is the mass of agent (in mg) collected on the sorbent tube or in the impinger over the
sampling interval T (in minutes), due to offgasing from the contaminated surface area A (in
2 2
cm ). Thus the units of OF are mg/cm /min.
The effectiveness with which the decontamination technology reduces the chemical vapor
offgasing will be calculated in a manner analogous to Equation 3, i.e.:
Eof = (OF0 - OFf)/OF0 • 100 (8)
where Eof is the percent efficacy for reducing the vapor offgas flux, and OF0 and OFf are the
contact transfer rates determined from the control and test coupons, respectively.
The agent vapor hazard is estimated based on the offgas flux, and assumed exposure time, room
ventilation, material surface area, and the hazard level for the vapor exposure to chemical agents.
Possible criteria for vapor exposure are listed below:'-4'1
• GD: 0.000001 mg/m3
• HD: 0.003 mg/m3
• VX: 0.000003 mg/m3.
8.3 Data Review
Records generated during verification testing will receive a one-over-one review before these
records are used to calculate, evaluate, or report verification results. These records may include
laboratory record books, completed data sheets, or reference method analytical results. This
review will be performed by a Battelle technical staff member other than the person who
originally generated the record. Testing staff will be consulted as needed to clarify any issues
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about the data records. The review will be documented by the person performing the review by
adding his/her initials and date to a hard copy of the record being reviewed. This hard copy will
then be returned to the Battelle staff member who generated or who will be storing the record.
8.4 Reporting
The efficacy calculations described in Section 8.2, the assessment of material damage, and other
observations during verification testing will be compiled in a verification report. The
verification report will present all the test data, supporting information on the measurement
methods, as well as the quantitative evaluation of the test results. The verification report will
briefly describe the ETV Building Decontamination Technology Center, and will describe the
procedures used in verification testing. The results of verification testing will then be stated
quantitatively, without comparison to any other technology, or any comment on the acceptability
of the technology's performance. The preparation of the draft verification report, the review of
the report by vendors and others, the revision of the report, final approval, and the distribution of
the report, will be conducted as stated in the Quality Management Plan (QMP) for this Center.(1)
Preparation, approval, and use of the Verification Statement summarizing the results of the
testing will also be subject to the requirements of that same QMP. For a technology undergoing
testing with both biological and chemical contaminants, separate verification reports will be
prepared on those two tests.
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9.0 HEALTH AND SAFETY
All participants in verification testing (i.e., Battelle, EPA, and vendor staff) will adhere to the
security, health, and safety requirements of HMRC and MREF. Vendor staff will train test
personnel in the use of their decontamination technology, but will not be the technology users
during the testing. For reasons of safety and controlled access at the West Jefferson facilities,
vendor staff may be able to observe some test procedures, but will not conduct any of the testing
activities.
9.1 Access
Access to restricted areas of the West Jefferson facilities will be limited to staff who have met all
the necessary training and security requirements. The existing access restrictions of the facilities
will be followed, i.e., no departure from standard procedures will be requested for verification
testing.
9.2 Potential Hazards
Verification testing conducted under this plan will involve the use of extremely hazardous
chemical and biological materials. Use of those materials must only be implemented in properly
certified surety facilities, capable of handling such materials safely.
In addition, surrogate materials used in such verification testing may also be toxic, and must be
used with appropriate attention to good laboratory safety practices.
9.3 Training
Because of the hazardous materials that will be involved in testing conducted under this plan,
documentation of proper training and certification of the test personnel is mandatory before any
testing takes place. The Battelle Quality Manager, or his counterpart at the West Jefferson
facilities, will assure that documentation of such training is in place for all test personnel before
allowing testing to proceed.
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9.4 Safe Work Practices
All visiting staff at the test facilities will be given a site-specific safety briefing prior to the start
of any test activities conducted under this plan. This briefing will include a description of
emergency procedures. Testing procedures must follow all specified safety practices at all times.
Any report of unsafe practices, by those involved in testing or by other observers, shall be
grounds for stopping testing until the appropriate facility safety officer and testing personnel are
satisfied that unsafe practices have been corrected.
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10.0 REFERENCES
1. Quality Management Plan (QMP) for the Technology Verification of Commercially
Available Methods for Decontamination of Indoor Surfaces Contaminated with Biological
or Chemical Agents, Version 1, prepared by Battelle, Columbus, Ohio, November 22,
2002.
2. Generic Verification Protocol for Technology Verification of Commercially Available
Methods for Decontaminating Indoor Surfaces Contaminated with Biological or Chemical
Agents, Version 1, prepared by Battelle, Columbus, Ohio, March 11, 2003.
3. Priority Threat Summary, prepared for EPA's Safe Buildings Program by Battelle,
Columbus, Ohio (draft), March 7, 2003.
4. Raber, E., Jin, A., Noonan, K., McGuire, R., and Kirvel, R.D. Decontamination Issues for
Chemical and Biological Warfare Agents: How Clean is Clean Enough? Int. J. Environ.
Health Res., 11, 128-148 (2001).
5. NBC Contamination Survivability Criteria for Military Equipment, Quadripartite
Standardization Agreement 747, Edition 1. 12 August, 1991.
6. Groenewold, G.S., Williams, J.M., Appellans, A.D., Gresham, G.L., Olson, J.E., Jeffrey,
M.T., and Rowland, B. Hydrolysis of VX on Concrete: Rate of Degradation by Direct
Surface Interrogation Using an Ion Trap Secondary Ion Mass Spectrometer, Environ. Sci.
Technol., 36, 4790-4794 (2002).
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APPENDIX A
TEST PERFORMANCE CONTROL SHEETS
TEST COUPON SAMPLE FORM
-------
TEST PERFORMANCE CONTROL SHEETS
CONTACT/EXTRACT TEST
DATE
OPERATOR
TRIAL#
ASSISTANCE
AGENT
RECORDER
SPIKE CONTROLS
MATERIAL
AGENT DROPS
EXTRACT
VOLUME
SAMPLE ID
GLASS
GLASS
GLASS
POSITIVE CONTROLS (NOT DECONTAMINATED)
MATERIAL
TYPE
REP
AGENT
APPLIEl
D
COUPON
EXTRACTED
ID
DROPS
TIME
TIME
VOLUME
A
1
A
2
A
3
B
1
B
2
B
3
C
1
C
2
C
3
D
1
D
2
D
3
-------
TEST SAMPLES
MATERIAL
REP
AGENT
HPV
CONTACT
EXTRACT
TYPE
APPLIED
GENERATION
TRANSFER
DROPS
TIME
START
END
TIME
VOLUME
TIME
VOLUME
A
1
A
2
A
3
B
1
B
2
B
3
C
1
C
2
C
3
D
1
D
2
D
3
A
NONE
B
NONE
C
NONE
D
NONE
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TEST PERFORMANCE CONTROL SHEETS
OFFGAS TEST
DATE
OPERATOR
TRIAL#
ASSISTANCE
AGENT
RECORDER
SPIKE CONTROLS
MATERIAL
AGENT DROPS
EXTRACT
VOLUME
SAMPLE ID
GLASS
GLASS
GLASS
TEST SAMPLES
MATERIAL
TYPE
REP
AGENT
APPLIED
HPV
GENERATION
OFFGAS SAMPLE
DROPS
TIME
START
END
INTERVAL
(TBD)
INTERVAL
(TBD)
INTERVAL
(TBD)
VOLUME
A
1
A
2
A
3
B
1
B
2
B
3
C
1
C
2
C
3
D
1
D
2
D
3
A
NONE
B
NONE
C
NONE
D
NONE
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E .£
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Initials &
Date
Comments/Observations (e.g., Color, Reflectivity, Apparent Roughness)
Heat-Shock Extract
from...
Time Spores Added
Tier No.
Test Coupon
Description
Test Coupon
Code
Sample ID
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APPENDIX B
VENDOR-SPECIFIC
TEST INFORMATION
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TEST/QA PLAN AMENDMENT
TEST/QA PLAN TITLE AND DATE: Test/OA Plan for Verification of Hydrogen Peroxide
Vapor Technologies for Decontaminating Indoor Surfaces Contaminated with Biological or
Chemical Agents. July 21. 2003
AMENDMENT NUMBER: 1-1
EFFECTIVE DATE: September 8, 2003
PART TO BE CHANGED/REVISED:
1) Page iv of the Table of Contents and the Appendix B section divider page in the back of the Test/QA Plan
2) Page 2, first paragraph of Section 1.3.1 and the Organization Chart on page 4
3) Organization Chart on Page 4
CHANGE/REVISION:
1) Remove references to Appendix B in the Table of Contents and remove the divider page referring to
Appendix B in the back of the plan report
2) Replace Dr. Thomas J. Kelly with Dr. Michael L. Taylor
3) Add Elisha N. Morrison as Biological Testing QA Coordinator under Zachary Willenberg
REASON FOR CHANGE:
1) Vendor/technology specific information will be included in the vendor/technology specific ETV
Verification Report, not the Test/QA Plan, to be consistent with ETV procedures.
2) Dr. Taylor will replace Dr. Kelly as Verification Testing Leader for the ETV Building Decontamination
Technology Center Project.
3) For The biological testing in the Medical Research and Evaluation Facility, Elisha Morrison will
coordinate QA activities under the overall direction of Zachary Willenberg.
APPROVED BY:
(SIGNATURES ON FILE)
Michael L. Taylor
Verification Testing Leader
09/08/03
Date
Zachary Willenberg
Battelle Quality Assurance Manager
09/09/03
Date
Required Distribution -
All individuals/organizations listed on distribution for the applicable test/QA Plan, including but
not limited to:
Battelle Program Management Verification Test Partners (if any)
Battelle Testing Staff EPA PO and TOPO
Battelle Quality Assurance Manager EPA Quality Staff
Vendors
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TEST/QA PLAN AMENDMENT
TEST/QA PLAN TITLE AND DATE: Test 0/A Plan for Verification of Hydrogen Peroxide Vapor
Technologies for Decontaminating Indoor Surfaces Contaminated with Biological or Chemical Agents.
July 21. 2003
AMENDMENT NUMBER: 1-2
EFFECTIVE DATE: November 18, 2003
PART TO BE CHANGED/REVISED:
1) Section 1.3.1 - Page 5, bullets 6 and 8 and concluding sentence
2) Section 3.2 - Page 12, first paragraph; Page 13, Table 3-1
3) Section 6.2.4 - Page 31, first paragraph
4) Appendix A - Test Coupon Sample Form
CHANGE/REVISION:
1) Remove bullets 6 and 8 and add the following concluding sentence: "William J. Ritter, the Facilities
Manager, reviews and approves data and records related to facility operation. Mr. Ritter will:
• Review and approve all data and records related to facility operation
• Provide input on facility procedures for the verification report"
2) Lines 4-8 of the first paragraph are amended to read: "The HMRC is an AR50-6 surety facility and is
certified to work with chemical surety material through a Bailment Agreement by the U.S. Army
Soldier biological and chemical command (SBCCOM). Battelle has demonstrated via...".
Table 3-1, The title is amended to read "Certification of HMRC" and the text of the table is amended
by changing the Bailment Agreement cited from No. DAAD13-H-03-0003 to No. DAAD13-03-H-
0003 (3 Mar 03 - 3 Mar 05).
3) Lines 1-2 of the first paragraph are amended to read: "After application of agents and surrogates to the
test coupons and completion of the drying or weathering time, the test coupons will be decontaminated
in the test chamber (Compact Glove Box). The control coupons will remain in the primary and
secondary containers within the Class II BSC and will not be placed in the test chamber at any time.
Each decontamination technology...".
4) Removed company name from before Parameters in the top right column.
REASON FOR CHANGE:
1) Reassign tasks from Testing Leader to Facilities Manager.
2) Update information on the HMRC certification.
3) Added a statement to clarify where the control coupons will be stored during decontamination.
4) Specific company name was removed so form can be used as a template.
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(SIGNATURES ON FILE)
APPROVED BY:
Michael L. Taylor
Verification Testing Leader
11/18/03
Date
Zachary Willenberg
Battelle Quality Assurance Manager
11/18/03
Date
Required Distribution -
All individuals/organizations listed on distribution for the applicable test/QA Plan, including but not
limited to:
Battelle Program Management
Battelle Testing Staff
Battelle Quality Assurance Manager
Verification Test Partners (if any)
EPA PO and TOPO
EPA Quality Staff
Vendors
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