&EPA
United States
Environmental Protection
Agency
TEST/QA PLAN FOR
Sabre Technical Services Chlorine Dioxide
Gas Generator
Office of Research and Development
National Homeland Security
Research Center
-------
-------
EPA/600/R-06/168
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page i of vi
EPA/Battelle Approval of
Test/QA Plan
For
EVALUATION OF SPORICIDAL
DECONTAMINATION TECHNOLOGIES
February 2005
(SIGNA TV RES ON FILE)
Original signed by:
Original signed by:
John C.S.Chang
John C.S. Chang, Ph.D.
Task Order Project Officer
U.S. EPA
04/04/05
Date
Eletha Brady-Roberts
Eletha Brady-Roberts
Quality Manager
U.S. EPA
04/01/05
Date
Original signed by:
Original signed by:
Thomas Kelly for
Karen Riggs
Program Manager
Battelle
03/30/05
Date
Zachary Willenberg
Zachary Willenberg
Quality Manager
Battelle
03/30/05
Date
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page ii of vi
Test/QA Plan
for
EVALUATION OF SPORICIDAL
DECONTAMINATION TECHNOLOGIES
February 2005
(SIGNA TV RES ON FILE)
Name David W. Skodack Signature David W. Skodack
Company Sabre Technical Services L.L.C.
Date 03/29/05
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page iii of vi
TABLE OF CONTENTS
Page
List of Acronyms v
Distribution List vi
A PROJECT MANAGEMENT 1
Al Technology Evaluation Organization 1
Al.l Battelle 2
A1.2 Vendors 5
A1.3 EPA 5
A1.4 Test Facility 6
A2 Problem Definition/Background 7
A3 Technology Evaluation Description and Schedule 8
A3.1 Applicability 8
A3.2 Scope 9
A3.3 Schedule 10
A4 Quality Objectives 10
A5 Special Training/Certification 12
A5.1 General Site Description 12
A5.2 Site Operations 13
A5.3 Training 14
A6 Documentation and Records 14
B MEASUREMENT AND DATA ACQUISITION 16
Bl Experimental Design 16
Bl.l General Test Design 16
B1.2 Scale of Testing and Testing Apparatus 17
B1.3 Test Surfaces 18
B1.4 Biological Agents and Surrogates 19
B1.5 Temperature and Relative Humidity Conditions 20
B1.6 Surface Damage 20
B2 Methods Requirements and Procedures 21
B2.1 Agents 21
B2.2 Coupon-Scale Testing 21
B2.2.1 Preparation of Test Materials 21
B2.2.2 Application of Biological Agents to Test Coupons 21
B2.2.3 Confirmation of Surface Application Density 22
B2.2.4 Application of the Decontamination Technology, Monitoring of
Test Procedures 22
B2.2.5 Determination of Decontamination Efficacy 22
B2.2.6 Observation of Surface Damage 24
B3 Quality Control Requirements 25
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page iv of vi
B4 Instrument Calibration and Frequency 25
B5 Data Acquisition Requirements 25
B6 Data Management 26
B6.1 Efficacy Calculations 26
B6.2 Statistical Analysis 26
C ASSESSMENT/OVESIGHT 28
Cl Assessments and Response Actions 28
Cl.l Technical Systems Audit 28
C1.2 Performance Evaluation Audits 28
C2 Reports to Management 28
D DATA VALIDATION AND USABILITY 30
E REFERENCES 31
APPENDIX
Detailed Descriptions of Decontamination Technologies and Operating Parameters for Efficacy
Evaluation
LIST OF FIGURES
Figure 1. Organization Chart for the Sporicidal Decontamination Technology
Evaluation 1
Figure 2. Compact Glove Box for the Sporicidal Decontamination Technology
Evaluation 18
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page v of vi
ANOVA
ATCC
BSC
BSL
C
CDC
CFR
CPU
C1O2
cm
d
EPA
FDA
h
ISO
L
mL
,iL
MREF
NHSRC
ppm
QA
QC
QMP
rpm
SD
SOP
TOPO
ISA
TTEP
UV
w
LIST OF ACRONYMS
two-way analysis of variance
American Type Culture Collection
biosafety cabinet
biosafety level
Celsius
Centers for Disease Control and Prevention
Code of Federal Regulations
colony forming unit
chlorine dioxide
centimeter
deep
U.S. Environmental Protection Agency
Food and Drug Administration
high
International Organization for Standardization
liter
milliliter
microliter
Medical Research and Evaluation Facility
National Homeland Security Research Center
parts per million
quality assurance
quality control
Quality Management Plan
revolutions per minute
standard deviation
standard operating procedure
Task Order Project Officer
technical systems audit
Technology Testing and Evaluation Program
ultraviolet
wide
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page vi of vi
DISTRIBUTION LIST
Ms. Karen Riggs
Mr. Zachary Willenberg
Dr. James Rogers
Battelle
505 King Avenue
Columbus, OH 43201-2693
Dr. Michael Taylor
Dr. Harry Stone
Battelle
10300 Alliance Rd, Ste 155
Cincinnati, OH 45242
Mr. Eric Koglin
USEPA National Homeland Security
Research Center
944 East Harmon Avenue
Las Vegas, NV 89119
Ms. Eletha Brady-Roberts
USEPA National Homeland Security
Research Center
26 W. Martin Luther King Drive
Cincinnati, OH 45268
Dr. John C.S. Chang
U.S. Environmental Protection Agency
U.S. EPAMailroom, E305-03
Research Triangle Park, NC 27711
Technology Vendor(s)
Mr. John Mason
Sabre Technical Services
17 Computer Drive East
Albany, NY 12205
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 1 of 31
A PROJECT MANAGMENT
Al TECHNOLOGY EVALUATION ORGANIZATION
The technology evaluation will be performed by Battelle under the direction of the U.S.
Environmental Protection Agency's (EPA) National Homeland Security Research Center
(NHSRC) through the Technology Testing and Evaluation Program (TTEP). This test/quality
assurance (QA) plan is based on a previously approved test/QA plan per the directive of the Task
Order Project Officer (TOPO). The organization chart in Figure 1 shows the individuals from
Battelle, the vendor, and EPA who will have responsibilities in the technology evaluation. The
responsibilities of these organizations and individuals are summarized in the following
subsections.
Battelle Management
Zachary Willenberg
Battelle Quality
Assurance Manager
Elisha Morrison
Biological Testing
QA Coordinator
Eric Koglin
EPA/NHSRC
Project Officer
Karen Riggs
Battelle TTEP
Manager
Eletha Brady-Roberts
NHSRC Quality
Assurance Manager
1
John Chang
EPA Task Order
Project Officer
Michael Taylor
Building Decontamination
Technology Area Leader/
Task Order Leader
James Rogers
Laboratory Test Coordinator
Medical Research and
Evaluation Facility (MREF)
Decontamination
Technology
Vendor
Representatives
Figure 1. Organization Chart for the Sporicidal Decontamination Technology Evaluation
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 2 of 31
Al.l Battelle
Dr. Michael Taylor is Battelle's Building Decontamination Technology Area Leader and
Task Order Leader for this technology evaluation. He will have overall responsibility for
ensuring that the technical, schedule, and cost goals established for testing and evaluation are
met, and that the procedures employed for testing are consistent with TTEP guidelines. Dr.
Taylor will serve as the primary interface for the TOPO. Dr. Taylor's responsibilities are to:
• Ensure that TTEP procedures are being followed.
• Select the appropriate laboratory or location for the evaluation.
• Prepare the draft test/QA plan and evaluation reports.
• Establish a test schedule.
• Revise this test/QA plan and evaluation reports in response to reviewers' comments.
• Keep the Battelle Program Manager informed of the progress and difficulties in
planning and conducting the evaluation.
• Coordinate with the Battelle Quality Assurance Manager for the performance of
technical and performance audits as required by Battelle or EPA Quality Management
staff.
• Have overall responsibility for ensuring that this test/QA plan is followed.
• Respond to any issues raised in assessment reports and audits, including instituting
corrective action as necessary.
• Establish a budget and schedule for the technology evaluation and direct the effort to
ensure that budget and schedule are met.
• Coordinate distribution of final test/QA plan and evaluation reports.
Ms. Karen Riggs is Battelle's TTEP Manager. As such, Ms. Riggs will:
• Maintain communication with EPA's NHSRC Project Officer on all aspects of the
program.
• Monitor adherence to budgets and schedules in this work.
• Provide the TOPO with monthly technical and financial progress reports.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page3 of 31
• Review the draft test/QA plan.
• Review the draft evaluation reports.
• Ensure that necessary Battelle resources, including staff and facilities, are committed
to the technology evaluation.
• Ensure that vendor confidentiality is maintained.
• Support Dr. Taylor in responding to any issues raised in assessment reports and
audits.
Mr. Zachary Willenberg is Battelle's Quality Assurance Manager for TTEP. As such,
Mr. Willenberg will:
• Review the draft test/QA plan.
• Maintain communication with EPA Quality Management staff for this program.
• Conduct a technical systems audit (TSA) at least once during the technology
evaluation.
• Audit at least 10% of the evaluation data.
• Prepare and distribute an assessment report for each audit.
• Verify implementation of any necessary corrective action.
• Notify Battelle's TTEP Manager to issue a stop work order if internal audits indicate
that data quality is being compromised. Notify the Task Order Leader if such an
order is issued.
• Provide a summary of the Q A/quality control (QC) activities and results for the
evaluation reports.
• Review the draft evaluation reports.
• Ensure that all quality procedures specified in this test/QA plan and in the Quality
Management Plan(1) (QMP) are followed.
Ms. Elisha Morrison will serve as Battelle's Biological Testing QA Coordinator and
assist Mr. Willenberg as necessary.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 4 of 31
Dr. James Rogers is Battelle's Laboratory Test Coordinator for this evaluation. His
responsibilities are to:
• Coordinate with vendor representatives to facilitate the performance of the
evaluation.
• Assist in preparation of the draft test/QA plan.
• Arrange for use of the test facility and establishment of evaluation schedules.
• Arrange for the availability of qualified staff to conduct the evaluation.
• Assure that the evaluation is conducted in accordance with this test/QA plan.
• Provide input into revision of this test/QA plan, evaluation report, and evaluation
statement in response to reviewers' comments.
• Update the Battelle TTEP Manager and Task Order Leader on progress and
difficulties in planning and conducting the evaluation.
• Coordinate with the Battelle Quality Assurance Manager for the performance of
technical and performance audits as required by Battelle or EPA Quality Management
staff.
A Medical Research and Evaluation Facility (MREF) Laboratory Facilities Coordinator
will review and approve data and records related to facility operation. This Facilities
Coordinator will:
• Review and approve all data and records related to facility operation.
• Provide input on facility procedures for the evaluation report.
• Provide requisite technical staff during the technology evaluation.
• Provide any safety training needed by Battelle, vendor, or EPA staff.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 5 of 31
Battelle technical staff will support Dr. Taylor in planning and conducting the technology
evaluation. These staff will:
• Ensure that the facility is fully functional prior to the times/dates needed in the
technology evaluation.
• Adhere to the requirements of the test/QA plan and the program WMP in carrying out
the technology evaluation.
• Support Dr. Taylor in responding to any issues raised in assessment reports and audits
related to facility operation.
A1.2 Vendors
Vendors of the sporicidal decontamination technologies will:
• Provide input for preparation of the draft test/QA plan.
• Review this test/QA plan and approve the current version prior to the evaluation of
their technology.
• Sign a Vendor Agreement specifying the respective responsibilities of the vendor and
of Battelle in the evaluation.
• Provide information on the quantitative response of their sporicidal decontamination
technology to aid in the planning of the evaluation.
• Provide the necessary equipment used for their sporicidal decontamination
technology for use in the technology evaluation.
• Train Battelle and/or test facility staff in the operation of their sporicidal
decontamination technology.
• Provide support, if needed, in use of their sporicidal decontamination technology
during testing.
• Review their respective draft evaluation report.
A1.3 EPA
Mr. Eric Koglin is the EPA/NHSRC Project Officer for the EPA contract with Battelle,
"Testing and Evaluation of Homeland Security-Related Technologies for the Measurement,
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 6 of 31
Sampling, Removal, and Decontamination of Chemical and Biological Agents" under which
TTEP has been established.
Dr. John Chang is the EPA TOPO for Task Order 1113. As such, Dr. Chang will:
• Have overall responsibility for directing the evaluation process.
• Review the draft test/QA plan.
• Approve the final test/QA plan and any subsequent versions.
• Review the draft evaluation reports.
• Oversee the EPA review process on the draft test/QA plan and evaluation reports.
• Coordinate submission of evaluation reports for final EPA approval.
Ms. Eletha Brady-Roberts is the NHSRC Quality Assurance Manager for the TTEP. As
such, Ms. Brady-Roberts will:
• Review the draft test/QA plan and any subsequent versions.
• Perform, at her option, one external TSA during the technology evaluation.
• Notify the EPA TOPO to issue a stop work order if an external audit indicates that
data quality is being compromised.
• Prepare and distribute an assessment report summarizing the results of the external
audit, if one is performed.
• Review the draft evaluation reports.
A1.4 Test Facility
The location for the technology evaluation described here will be Battelle's laboratories
in Columbus and West Jefferson, Ohio. The Columbus facilities to be used are chemical
laboratories equipped for safe handling of a wide variety of chemicals. The MREF, located in
West Jefferson, has chemical and biological surety agent laboratories certified for use of
chemical and biological warfare agents. Other test facilities could be used depending on the
availability and capability of the facilities.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 7 of 31
A2 PROBLEM DEFINITION/BACKGROUND
Among its responsibilities related to Homeland Security, the 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 NHSRC to manage, coordinate, and support a wide variety of homeland
security research and technical assistance efforts. The NHSRC is conducting tests to evaluate
the performance of commercially-available products, methods, and equipment for
decontamination of porous and non-porous indoor surfaces contaminated with biological or
chemical agents.
The purpose of this testing is to generate objective performance data that can be used by
building and facility managers, first responders, groups responsible for building
decontamination, and other technology buyers and users to make informed purchase and
application decisions. All potential users need unbiased, high-quality, objective third-party data
and information in order to assess how well the available decontamination tools will meet their
performance objectives while protecting human health and the environment. All testing and
evaluation conducted through the TTEP is under the direction of EPA and is subject to the TTEP
QMP. In performing each test, Battelle will follow the general procedures described in the
QMP, and develop a separate test/QA plan that is specific for the type of decontamination
technology being tested. This particular test/QA plan has been prepared for testing and
evaluation of decontamination technologies that use gaseous or sporicidal decontamination
agents [e.g. chlorine dioxide (C1O2), hydrogen peroxide vapor, formaldehyde].
The objective of this test/QA plan is to describe laboratory test procedures that will be
implemented to determine the efficacy of sporicidal technologies for removing or inactivating
biological agents or surrogates on a range of representative indoor surfaces. This test/QA plan is
specifically focused on decontamination of indoor surfaces typical of those found in a public
building with the ultimate goal of providing technologies for restoring the building to a usable
state following a terrorist attack. Decontamination of personnel or large equipment items (e.g.
manufacturing equipment) is not covered in this test/QA plan. Decontamination technology
testing and evaluation are being performed to generate data indicative of the technology
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 8 of 31
performance or efficacy. For the evaluation conducted under this test/QA plan, quantitative
assessment of performance is accomplished by sampling and analysis of contaminants before and
after the implementation of the decontamination technology. The performance parameters to be
evaluated in the evaluation under this test/QA plan are discussed in the Appendix.
One or more biological agents (e.g., spores; vegetative cells, biotoxins) may be released
inside a building during a terrorist attack. The highly persistent biological warfare agent,
Bacillus anthracis Ames spores, was selected for this evaluation.
Indoor surfaces (e.g., carpet, laminate, wood) representing those found in a typical office
building have been selected for use in evaluating the decontamination technology. The indoor
surfaces selected include both porous and non-porous materials (see Section B1.3).
A3 TECHNOLOGY EVALUATION DESCRIPTION AND SCHEDULE
The overall objective of the evaluation called for under this test/QA plan is to determine
the efficacy of the sporicidal decontamination technologies for removing/inactivating biological
agents in or on typical indoor surfaces. Evaluation of each technology will be accompanied by
careful monitoring of dwell time, decontamination agent concentration, temperature, relative
humidity and other parameters that may impact decontamination efficacy.
A3.1 Applicability
This test/QA plan focuses on the evaluation of commercially available technologies for
decontaminating indoor surfaces found in a typical office building or subway. This plan
specifically focuses on building decontamination in the context of use by personnel responsible
for decontamination after a terrorist attack. Toxic industrial chemicals, chemical warfare agents
and/or biological warfare agents (including toxins) may pose a threat in the building
contaminated by a terrorist attack. This plan focuses on the evaluation of technologies that are
potentially applicable for decontaminating indoor surfaces contaminated with biological warfare
agents (or toxins).
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 9 of 31
Technology evaluation testing requires a quantitative basis for establishing the
performance of the tested technologies. For this evaluation, the performance of each of the
decontamination technologies will be evaluated by comparing the amount of biological agent
remaining on the indoor surface (carpet, wallboard, etc.) after decontamination with the amount
of biological agent that was added to the indoor surface prior to decontamination.
A3.2 Scope
The overall objective of the technology evaluation described in this test/QA plan is to
evaluate the performance of decontamination technologies using biological agents (including
toxins) and surrogates under a range of realistic conditions. Testing may be conducted over
ranges of temperature and relative humidity representing those that might be encountered in a
decontamination situation in a building environment.
The performance parameters on which the decontamination technologies will be
evaluated under this plan include:
• Log kill or efficacy [the logarithm (base 10) of the number of colony-forming units
(CPUs) removed/destroyed by applying the decontamination technology].
• Surface damage caused by the decontamination technology.
The evaluation to be conducted under this plan is limited to detection of biological
warfare agents (or surrogates) or toxins in or on individual samples (test coupons) of indoor
materials. Testing will be conducted in two phases. The initial phase will take place in a naive
area (not a biological safety laboratory) and the second phase will take place in a biosafety level
(BSL) 3 area.
In the initial phase, the decontamination technology will be coupled with the test
chamber, associated monitoring devices will be installed in the test chamber and the vendor staff
will train the MREF staff regarding use of the decontamination technology. In addition, the
MREF staff will verify that all sub-systems comprising the testing apparatus are in working
order.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 10 of 31
The second phase of the testing entails evaluating the performance of the
decontamination technology using live agent. This evaluation will be performed in a BSL-3 in
the MREF.
A3.3 Schedule
The evaluation described in this test/QA plan is expected to commence within two weeks
after this test/QA plan has been approved. It is anticipated that four weeks will be required to
complete all testing for a single sporicidal technology. This schedule is predicated upon the
vendor shipping the decontamination technology to Battelle and training Battelle personnel in the
use of the equipment in accordance with the overall testing schedule.
A4 QUALITY OBJECTIVES
The performance parameters to be evaluated under this test/QA plan include:
• Quantitative assessment of decontamination efficacy of sporicidal technologies.
CPUs in extracts of control test coupons are determined using standard plating
techniques. Plate counts for three controls (inoculated with 108 spores per coupon but
not decontaminated) is determined and each of these values is used to calculate the
value N/N' where N is the number of CPUs found on the control coupon and N' is the
number of CPUs found on the decontaminated coupon. The log [10] of each of these
is calculated and the three logs are averaged to obtain a mean log kill ± standard
deviation (SD). The log kill value is also efficacy discussed in Section Bl.l. The log
kill or efficacy value is used as an indication of the overall effectiveness of the
decontamination technology. The closer the efficacy value is to 108 (the number of
spores applied to the test coupon) the more effective or efficacious the treatment.
• Qualitative assessment of residual biological agent and surrogate spores on test
surfaces following decontamination and extraction. Following the extraction to
determine the efficacy value quantitatively, test coupons will be immersed in liquid
growth medium contained in individual vials, cultured at the respective temperatures,
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 11 of 31
and bacterial growth in the vials will be observed visually at one and seven days after
immersion. If the growth medium appears cloudy, then the conclusion is drawn that
viable spores remained on the decontaminated samples following extraction. Growth
could arise from spores added to the test coupon or reflect growth of indigenous
organisms. Spores or other organisms giving rise to growth in the nutrient medium
will be identified by standard culture techniques. A small sample of each positive
culture will be streak plated onto tryptic soy agar plates and cultured overnight. The
streak plates will provide information as to whether the organisms present are the
same as those inoculated on the coupons, or other distinct microorganisms
Qualitative assessment of spore strips and biological indicators. Following
implementation of the decontamination technology the spore strips and indicators are
placed in growth medium. The growth medium is examined at one and seven days
post-decontamination for evidence of viable spores (medium becomes cloudy). No
cloudiness indicates no viable spores which indicates 100% kill or a log kill of 106.
The biological indicators and spore strips have been used in large-scale
decontaminations to assess the completeness of decontamination. However, the
spores on the biological indicators and spore strips are not B. anthracis spores but
surrogates. In addition, the spore strips and biological indicators are more amenable
to decontamination (biological indicators are a metal disc, spore strips are filter
paper). The results for biological indicators and spore strips provide a means of
comparing the effectiveness of the decontamination technology with other results
obtained using the same or other decontamination technologies.
Changes in appearance of representative indoor surfaces. Each test coupon is
visually examined before and after application of the decontamination technology to
assess whether or not the test coupon material exhibits physical changes (loss or
change of color etc.). This information provides a general indication of how
"caustic" the decontamination technology is and whether or not the contents of, for
example, a subway station would be damaged or destroyed during decontamination.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 12 of 31
Quantitative determinations in this study do not involve the use of analytical
measurement devices. Rather, bacterial colonies will be enumerated manually and recorded. All
other determinations will be qualitative. Specific information regarding the individual
decontamination technologies is included in the Appendix.
A5 SPECIAL TRAINING/CERTIFICATION
These tests are expected to be conducted at the Battelle facility in West Jefferson, Ohio.
That facility is described below. Alternative facilities could also be used, provided those
facilities meet all the requirements for safety, security, and testing capability established by this
test/QA plan.
A5.1 General Site Description
Evaluation of sporicidal decontamination technologies will be conducted at Battelle's
MREF located in West Jefferson, Ohio, near Battelle's headquarters in Columbus, Ohio. The
following section describes the MREF biofacility. The evaluation will be performed in
accordance with Battelle's facility-specific methods and standard operating procedures (SOPs)
that are cited where appropriate throughout this test/QA plan.
The MREF specializes in research, development, testing, and evaluation 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. This facility maintains state-of-
the-art equipment, and professional and technical staffing expertise to safely conduct testing and
evaluation of hazardous biological materials under the Food and Drug Administration's (FDA)
Good Laboratory Practices Guidelines (21 CFR Part 58). The MREF operates in compliance
with all applicable Federal, state, and local laws and regulations, including U.S. Army
regulations and is routinely inspected by personnel from the appropriate government agency.
Battelle operates the MREF in compliance with requirements contained in 32 CFR 626 and 627,
Biological Defense Research Programs. The MREF facilities are ISO 9001 certified, accredited
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 13 of 31
by the American Association for the Accreditation of Laboratory Animal Care, and inspected
and compliant with the U.S. Department of Agriculture, 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, 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. The MREF
fully complies with all applicable U.S. Army Regulations, and Federal Government and State of
Ohio regulations to conduct and support research, development, testing and evaluation studies
using highly toxic chemical and pathogenic biological materials. The MREF is licensed to ship,
receive, and handle select agents, as defined by the Centers for Disease Control and Prevention
(CDC).
Testing outlined in this test/QA plan will be performed in the MREF BSL-3 facility,
which was completed in 1995 and expanded to 31,000 square feet in 2002. The containment
area within the facility is designed to meet or exceed the BSL-3 facility guidelines published by
the CDC and National Institute of Health entitled Biosafety in Microbiological andBiomedical
Laboratories (4th edition, 1999). Included are seven BSL-3 microbiology laboratories that
contain multiple Class III biosafety cabinets (BSCs) and two autoclaves. Additional laboratories
within this area include multiple microbiology laboratories equipped with Class II BSCs. Test
procedures at the MREF are governed by established SOPs that are specified by facility, number,
and title.
A5.2 Site Operations
Battelle operates the MREF in compliance with all applicable Federal, state, and local
laws and regulations, including U.S. Army Regulations and the CDC. Battelle's facilities are
certified through inspection by personnel from the appropriate government agency. Battelle's
MREF is certified to work with both live and surrogate agents including bacterial endospores
(e.g. B. anthracis), vegetative bacteria, and viruses. Additionally, the MREF is ISO 9001
certified, performs work under this ISO standard, and is monitored by regular outside ISO
quality inspections.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 14 of 31
A5.3 Training
Because of the hazardous materials involved in this technology evaluation,
documentation of proper training and certification of the test personnel is mandatory before
testing takes place. The Battelle Quality Assurance Manager, or a designee, must assure that
documentation of such training is in place for all evaluation personnel before allowing evaluation
to proceed.
All participants in this evaluation (i.e., Battelle, EPA, and vendor staff) will adhere to the
security, health, and safety requirements of the Battelle facility in which testing will be
performed. Vendor staff will train Battelle evaluation staff in the use of their decontamination
technology, but will not be the technology users during the evaluation. To the extent allowed by
the test facility, vendor staff may observe, but may not conduct, any of the technology evaluation
activities identified in this test/QA plan.
Access to restricted areas of the test facility will be limited to staff who have met all the
necessary training and security requirements. The existing access restrictions of the test facility
will be followed, i.e., no departure from SOPs will be needed for this evaluation. All visiting
staff at the test facility will be given a site-specific safety briefing prior to the start of any
technology evaluation activities. This briefing will include a description of emergency operating
procedures and the identification, location, and operation of safety equipment (e.g., fire alarms,
fire extinguishers, eye washes, exits). Evaluation procedures must follow all safety practices of
the test facility at all times. Any report of unsafe practices in this evaluation, by those involved
in the evaluation or by other observers, shall be grounds for stopping the evaluation until the
Quality Assurance Manager and testing personnel are satisfied that unsafe practices have been
corrected.
A6 DOCUMENTATION AND RECORDS
Documentation of training related to technology testing, field testing, data analysis, and
reporting is maintained for all Battelle technical staff in training files at their respective
locations. The Battelle Quality Assurance Manager may verify the presence of appropriate
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 15 of 31
training records prior to the start of testing. Battelle will document training by the vendor with a
form signed by the vendor. Battelle technical staff will have a minimum of a bachelor's degree
in science/engineering or have equivalent work experience.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 16 of 31
B MEASUREMENT AND DATA ACQUISITION
Bl EXPERIMENTAL DESIGN
Bl.l General Test Design
This test/QA plan specifies procedures for bench-scale testing to evaluate the
performance of sporicidal decontamination technologies under specified operating conditions
and ambient conditions for decontaminating small pieces (i.e. test coupons) of indoor materials
to which biological agents and surrogates have been added.
Evaluation of the efficacy of a particular sporicidal technology for decontaminating a
particular material will be accomplished by determining the differential mean reduction of viable
agent or surrogate under an experimental treatment compared to mean reduction in the absence
of the treatment (control). Treatments will be defined in terms of viable agent or surrogate,
decontamination technology (or lack thereof) and operational use of the technology (e.g.,
concentrations, dwell times, flow/vent rates), and ambient conditions (e.g., temperature,
humidity). Decontamination efficacy will be assessed by comparing the number of viable
organisms remaining after decontamination with the number initially applied to the test surface.
Efficacy will be expressed as the log (base 10) of result of dividing the mean number of viable
organisms found on the control by the number of viable organisms found on a treated sample.
Triplicate samples (e.g., 3 carpet, 3 wood, etc.) are subjected to the decontamination technology
and an efficacy value for each sample is calculated and these three values are used to calculate a
mean and SD.
In advance of each efficacy evaluation, a null hypothesis (Ho) will be formulated for the
mean differential performance of the experimental treatment and control data:
HO '• Rrreatment ~ Rcontrol > C
HA '• ^-Treatment ~ -^Control ^ C
Where:
is the mean removal for the treatment group.
is the mean removal for the control group.
c is a constant that could be zero if desirable to identify any significant removal or some
threshold value for demonstrating minimum efficacy (e.g., 99.9999% removal).
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 17 of 31
The experimental treatment and controls will be defined according to desired planned
comparisons. For any particular material, the planned comparisons may include:
• Efficacy of a decontamination technology under selected environmental conditions for a
biological agent compared to a surrogate(s).
• Efficacy of a particular decontamination technology under selected environmental
conditions and a particular biological agent for each of several different configurations of
the technology (e.g., different concentrations, dwell times) or between use and absence of
the technology.
After obtaining the experimental treatment and control data, a statistical comparison will be
conducted. If sufficient evidence is found (at 95% confidence) that the experimental treatment
mean removal exceeds the control mean removal above the applicable threshold "c", the
treatment will be concluded to be efficacious.
A parallel sensitivity analysis will be conducted to determine, based on the variability of
the data collected, how large a true difference in efficacy would be highly likely (90%) to have
been identified as statistically significant by this hypothesis test.
Throughout the evaluation, the test coupons will be randomly assigned to control and
experimental groups. Each decontamination technology will be applied in a manner consistent
with the manufacturer's recommendations. The technology vendor will provide the equipment
and training regarding application of their technology.
The statistical approaches for these analyses are discussed in Section B6.2.
B1.2 Scale of Testing and Testing Apparatus
The parameters listed above will be evaluated during bench-scale testing in the
laboratory. A decontamination test chamber, a Compact Glove Box 830-ABC (Plas Labs, Inc.,
Lansing, MI; see Figure 2) will be used for exposing the test coupons to the decontamination
technology. This test chamber has dimensions of 71 cm w x 59 cm d x 74 cm h (28" x 23" x
29") and outer dimensions of 110 cm w x 61 cm d x 79 cm h (43" x 24" x 31"). The test
chamber has a total volume of 317L (11.2 cubic feet). The test chamber also has a top opening
of 43 cm x 58 cm (17" x 23") and an attached transfer chamber that is 30 cm (12") long and an
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 18 of 31
inner diameter of 28 cm (11"). Glove ports are available for working in the test chamber. The
test chamber may need to be modified, in accordance with the vendor, to accommodate the
decontamination technology. The decontaminant (fumigant, foam, liquid, gel) will be directed
from the vendor's system into the test chamber under the conditions (dwell time, decontaminant
concentration, temperature, and humidity) specified by the vendor. The performance tests will
use test coupons that are approximately 1.9 cm w x 7.5 cm long (%" x 3"); multiple coupons of
each indoor material will be contaminated with the agent/surrogate, placed into the test chamber
and then treated with the decontamination technology. Blank (i.e., uncontaminated) and control
(i.e., contaminated but not decontaminated) coupons will also be prepared for each test material,
and results obtained for these coupons will be utilized along with the data resulting from the
analyses of post-treatment samples to calculate decontamination efficacy. This evaluation
methodology comprises a highly controlled, reproducible approach to assess decontamination
efficacy, while simulating a realistic, small-scale application of the decontamination technology.
lompact
Glove Box
ti.W-AHC
Figure 2. Compact Glove Box for the Sporicidal Decontamination Technology Evaluation
B1.3 Test Surfaces
A subset of the various structural, decorative, and functional surfaces that may be found
inside an office building will be used for evaluating sporicidal decontamination technologies.
The surface materials to be used include both non-porous and porous surfaces representing a
variety of materials. Test coupons (typically measuring 1.9 cm x 7.5 cm) will be prepared from
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 19 of 31
larger pieces of stock material. The representativeness and uniformity of the test materials are
critical attributes to assure reliable evaluation results. Representativeness means that the
materials used are typical of such materials used in buildings in terms of quality, surface
characteristics, structural integrity, etc. Uniformity means that all test pieces are essentially
equivalent for evaluation purposes. Representativeness will be assured by selection of test
materials that meet industry standards or specifications for indoor use, and by obtaining those
materials from appropriate suppliers. Uniformity will be maintained by obtaining a large enough
quantity of material that multiple test samples can be obtained with presumably uniform
characteristics (e.g., test coupons will be cut from the interior rather than the edge of a large
piece of material), or by using standardized coupons where available. The test surfaces that will
be used are listed below with their corresponding sample identification codes in parentheses:
• Painted (latex, semi-gloss) concrete cinder block (PC)
• Painted (latex, flat) wallboard paper (PW)
• Decorative laminate (DL)
• Galvanized metal ductwork (GM)
• Glass (GS)
• Bare pine wood (BWD)
• Industrial carpet (1C)
B1.4 Biological Agents and Surrogates
The biological agent to be used in the evaluation under this test/QA plan has been
selected based on an evaluation of potential threats to buildings.(3) The evaluation considered
availability, lethality, potential delivery pathways and persistence of potential agents.
The biological agent used in evaluating the sporicidal decontamination technology will
be B. anthracis Ames strain spores. The biological surrogates will be used to establish
correlations between the decontamination efficacy of surrogates and agents. To provide
correlations with the B. anthracis results, the surrogates Bacillus subtilis (ATCC 19659) and
Geobacillus stearothermophilus (ATCC 12980) will be used. B. anthracis Ames strain spores
and surrogate spores will be prepared and characterized according to MREF SOPs.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 20 of 31
Both spores (B. subtilis and G. stearothermophilus) have commonly been used as surrogates for
B. anthracis in decontamination technology testing. The G. stearothermophilus surrogate
exhibits comparatively high resistance to various sporicidal decontaminants. The B. subtilis
(ATCC 19659) surrogate is the most commonly used surrogate for B. anthracis.
Two types of biological indicators will be used under this test/QA plan. A commercial
spore strip of the same spore type [B. subtilis var niger (B. atrophaeus ATCC 9372) on paper
backing manufactured by Raven Biological Laboratories] as those used during decontamination
of U.S. Postal Service facilities contaminated with B. anthracis will be used. In addition,
biological indicators containing B. stearothermophilus and B. subtilis (Apex Laboratories, Inc.)
will be included. Each of these biological indicators typically contains a spore population of
approximately 106 spores on a stainless steel disc packaged in a Tyvek® envelope.
B1.5 Temperature and Relative Humidity Conditions
Each sporicidal decontamination technology may require different temperature and
humidity conditions in the space to be decontaminated. The temperature and the relative
humidity will be controlled and monitored during the decontamination process. Specific
operating parameters and conditions for each sporicidal decontamination technology are
specified in the Appendix.
B1.6 Surface Damage
The effect of the decontamination technology on the test coupons will be evaluated
during the efficacy evaluation procedure. Before and after decontamination of the test coupons,
the appearance of the decontaminated coupons will be visually inspected, and any obvious
changes in the color, reflectivity, and apparent roughness of the coupon surfaces will be recorded
in the evaluation. This comparison will be performed for each of the test materials, before
extraction or sampling of the decontaminated test coupons.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 21 of 31
B2 METHODS REQUIREMENTS AND PROCEDURES
B2.1 Agents
B. anthracis (Ames), B. subtilis (ATCC 19659) and G. stearothermophilus (ATCC
12980) spores will be prepared according to established MREF procedures.(4'5) Working stock
suspensions of each spore type will be prepared at a target concentration of approximately IxlO9
CFU/mL.
B2.2 Coupon-Scale Testing
B2.2.1 Preparation of Test Materials
Each of 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 prior to inoculation with the biological agent or surrogates
and any surface anomalies will be recorded. Specifications of each test coupon will be recorded.
On each evaluation day, each coupon will be assigned a unique identifier code by the evaluation
staff for traceability. Prior to the application of the biological agent or surrogate, the surface of
each test coupon will be wiped with 70% isopropanol or ethanol to kill endogenous
microorganisms. This is intended to minimize contamination by microorganisms other than
those being evaluated. To ensure further cleanliness and prevent contamination of test surfaces,
sterile technique will be exercised during all phases of handling the test coupons.
B2.2.2 Application of Biological Agents to Test Coupons
Application of biological agent/surrogates to test coupons will be performed in a BSC III
according to established MREF procedures.(6) Test coupons will be placed lying flat in the
cabinet and contaminated at challenge levels of approximately 1 x 108 CFU per coupon. A 100
|iL aliquot of a stock suspension (approximately 1 x 109 CFU/mL) of spores will be dispensed
using a micropipette as small droplets across the surface of the test coupon. After contamination
with biological agent or surrogate suspension, the test coupons will remain overnight and
undisturbed in the BSC III to dry.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 22 of 31
B2.2.3 Confirmation of Surface Application Density
To confirm the application density of biological agents and surrogates, the respective
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 B2.2.5.
B2.2.4 Application of the Decontamination Technology, Monitoring of Test Procedures
One day following application of biological agent or surrogate, inoculated test coupons
intended for decontamination (including one blank) will be transferred into the test chamber that
has been coupled with the decontamination technology. The inoculated control coupons (not
decontaminated) and one blank will be left undisturbed in the BSC. The decontamination
technology will be applied in accordance with the vendor's instructions. The concentration of
the gaseous decontaminant, dwell time, relative humidity, and temperature will be controlled and
monitored as described in the Appendix. Following decontamination, the test chamber will be
cleared using the vendor-supplied method for neutralization of decontamination chemicals.
B2.2.5 Determination of Decontamination Efficacy
The efficacy of a sporicidal technology for neutralizing/inactivating biological agents on
indoor surface materials will be determined. For building decontamination, the amount of
biological agent that remains following decontamination needs to be ascertained since residual
agent could present a potential health risk for building occupants. The performance or efficacy
of the sporicidal decontamination technology will be assessed by comparing the number of
viable organisms remaining after decontamination with the number actually added to test
coupons. For biological agents, there is currently no determined safe level for remaining residual
organisms;(2) however, a traditional approach for qualifying decontamination performance is the
assessment of growth of biological indicators pre-positioned inside the space being fumigated.
Typically, biological indicators contain spore loads of approximately 1 x 106; therefore, complete
neutralization of these biological indicators results in a 6-log kill. As stated above there is no
officially recognized clean level for building decontamination; however, biological indicators
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 23 of 31
(used in other decontamination efforts) will also be used for this evaluation as a means for
evaluating the efficacy of the sporicidal decontamination technology.
The decontaminated, control, and blank coupons will be placed individually in a sterile
50 mL conical vial to which 10 mL of sterile phosphate-buffered saline containing 0.1% Triton
X-100 has been added. The purpose of the Triton X-100 is to minimize clumping of spores. For
spore extraction, the tubes will be agitated on an orbital shaker for 15 minutes at approximately
200 rpm at room temperature. Each tube will then be heat-shocked for 1 hour at 60° C to 65° C
(for B. anthracis and B. subtilis) or 90° C to 95° C (for G. stearothermophilus) to kill vegetative
bacteria. Following the heat-shock, 1.0 mL of the extract will be removed and a series of
dilutions through 10"7 will be prepared in sterile water. An aliquot (100 jiL) of the undiluted
extract and each serial dilution will be plated onto tryptic soy agar plates in triplicate, allowed to
dry, and incubated overnight at 35° C to 37° C for B. anthracis and B. subtilis and at 55° C to 60°
C for G. stearothermophilus. Plates will be enumerated within 18 to 24 hours of plating
as described in References 7 and 8. The number of CFU/mL will be determined by multiplying
the average number of colonies per plate by the reciprocal of the dilution. Data will be expressed
as mean + SD of the numbers of CPUs observed. To calculate the percent recovery of spores
following treatment, the calculated number of spores remaining on the decontaminated coupons
will be divided by the calculated number of spores on the control coupons that were not subject
to decontamination.
Decontamination efficacy will be calculated as the log reduction (mean ± SD) in viable
organisms achieved by the decontamination technology. Efficacy (E) for biological agents or
surrogates will be calculated as
E = log (N/N')
Where N is the number of viable organisms recovered from the control coupons (i.e., those not
subjected to decontamination), and N' is the number of viable organisms recovered from the test
coupons after decontamination.
A separate efficacy calculation will be made for each of the surface materials for the
biological agent and surrogates. Percent recovery (mean ± SD) will be calculated for each type
of test material inoculated with each biological agent/surrogate by dividing the number of
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 24 of 31
biological organisms in the treated sample by the number of biological organisms in the control
(non-decontaminated). 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.
Based on previous decontamination studies, it is assumed that 100% recovery of spores
from the inoculated test coupons will not be achieved; therefore, viable spores may remain on the
test coupons. A qualitative assessment of these spores will be performed to determine whether
viable spores remain on the decontaminated test coupons. Following the extraction process
described above, each coupon will be transferred into a sterile 50 mL conical tube containing 20
mL of tryptic soy broth culture medium. These vials will be cultured at the appropriate
temperature for B. anthracis or surrogates to encourage viable spore germination and subsequent
proliferation of vegetative bacteria. At one and seven days post-decontamination the tubes will
be visually assessed qualitatively for viability. A cloudy culture medium will indicate "growth"
of viable spores, while clear culture medium will indicate "no growth." In the seven day samples
where growth is observed, a loop of the culture medium will be cultured on tryptic soy agar
plates using a standard streak plate technique. This culturing on tryptic soy agar plates will
provide a means to determine whether the observed growth in the liquid culture medium is due to
the B. anthracis or surrogate applied to the coupon, a contaminant, or both.
The biological indicators and spore strips will be cultured in tryptic soy broth and
assessed qualitatively at one and seven days post-decontamination for growth or no growth.
However, further culturing of any positive samples on tryptic soy agar plates will not be
performed.
B2.2.6 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 performed immediately after
completion of the decontamination process but before post-decontamination sampling to assess
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 25 of 31
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, and roughness will be assessed qualitatively and
observations will be made by the evaluation staff and recorded.
B3 QUALITY CONTROL REQUIREMENTS
Quantitative standards do not exist for biological agents and surrogates. The
confirmation procedure, controls, blanks, and method validation efforts will be the basis of
support for biological evaluation results.
B4 INSTRUMENT CALIBRATION AND FREQUENCY
The equipment needed for the evaluation will be maintained and operated according to
the quality requirements and documentation of the evaluation facility. Relative humidity and
temperature in the test chamber will be monitored using a NIST-traceable digital thermometer -
hygrometer. The accuracy of the instrumentation used to monitor the decontamination reagent
(e.g. fumigant) will be verified as indicated in the Appendix.
B5 DATA ACQUISITION REQUIREMENTS
There are no data needed for this project implementation that are obtained from non-
measurement sources such as computer databases, programs, literature files or historical
databases.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 26 of 31
B6 DATA MANAGEMENT
Data acquisition during the evaluation includes proper recording of the procedures used
in the evaluation to assure consistency in the evaluation and adherence to this test/QA plan;
documentation of sampling/evaluation conditions; recording observations regarding the
condition of the surface of each coupon before and after the decontamination process; and
recording of efficacy results and evaluation conditions. Data acquisition will be carried out by
the Battelle testing staff manually and recorded immediately in a consistent format throughout all
evaluations. All written records will be in ink and any corrections to recorded data will be made
with a single line through the original entry. The correction will then be entered, initialed, and
dated by the person making the correction. Any non-obvious correction will include a reason for
the correction. Strict confidentiality of evaluation data will be maintained.
B6.1 Efficacy Calculations
For biological agents and surrogates, decontamination efficacy will be calculated as
(described in Section B2.2.5) the reduction in viable organisms achieved by the technology.
B6.2 Statistical Analysis
For each material and species combination, calculating log reduction values will result in
a total of 63 efficacy values (that is three coupons for each of seven materials analyzed in
triplicate). In cases where no viable colonies remain after decontamination, one colony will be
assumed to be present for the purpose of this calculation. A two-way analysis of variance
(ANOVA) model with main effects for each test organism and test material and interactions will
be fitted to the efficacy data. This model will be used to compare each mean to zero, compare
each surrogate to a selected organism, for example B. anthracis (for a specific material), and
compare each surrogate to a selected organism for porous and non-porous materials. T-tests or
statistical contrasts will be used for the comparisons, with no adjustment for multiple
comparisons. The ANOVA model is fitted using the SAS® (Version 8.2) GLM procedure.
The evaluation results will be compiled in a report. The report will briefly describe
TTEP and evaluation procedures as well as all evaluation data and observations. The preparation
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 27 of 31
of the draft report, review of the draft report, the revision of the draft report, final approval, and
the distribution of the final report, will be conducted as stated in the QMP.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 28 of 31
C ASSESSMENT/OVERSIGHT
Cl ASSESSMENTS AND RESPONSE ACTIONS
Battelle's Quality Assurance Manager will audit at least 10% of the evaluation data. The
Quality Assurance Manager will trace the data from initial acquisition, through reduction and
statistical comparisons, to final reporting. All data calculations will be checked.
Cl.l Technical Systems Audit
Battelle's Quality Assurance Manager or designee will perform one TSA during the
evaluation. The TSA is to ensure the evaluation is performed in accordance with the TTEP QMP
and the test/QA plan and that QA/QC procedures are implemented. The Quality Assurance
Manager may review evaluation methods, compare test procedures to those specified in this
test/QA plan, and review data acquisition and handling procedures. The Quality Assurance
Manager will prepare a TSA report and the findings must be addressed either by modifications of
test procedures or by documentation in the evaluation records and final report. At EPA's
discretion, EPA QA staff may also conduct an independent on-site TSA during the evaluation.
The EPA TSA findings will be communicated to evaluation staff at the time of the audit, and
documented in a TSA report. These findings must be addressed as stated above.
C1.2 Performance Evaluation Audits
No performance evaluation audit will be performed for biological agents and surrogates,
as quantitative standards for these biological materials do not exist. The confirmation procedure,
controls, blanks, and method validation efforts will be the basis of support for biological
evaluation results.
C2 REPORTS TO MANAGEMENT
Each assessment and audit will be documented in accordance with the QMP. Assessment
reports will include the following:
• Identification of any adverse findings or potential problems.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 29 of 31
• Space for response to adverse findings or potential problems.
• Possible recommendations for resolving problems.
• Citation of any noteworthy practices that may be of use to others.
• Confirmation that solutions have been implemented and are effective.
During the course of any assessment or audit, the Quality Assurance Manager will identify to the
technical staff performing experimental activities any immediate corrective action that should be
taken. If serious quality problems exist, the Quality Assurance Manager is authorized to stop
work. Once the assessment report has been prepared, the Building Decontamination Technology
Area Leader or Task Order Leader 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 Assurance Manager will ensure that follow-up corrective action has been taken.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 30 of 31
D DATA VALIDATION AND USABILITY
Records generated during the evaluation will receive a QC/technical review before these
records are used to calculate, evaluate, or report results. This review will be performed by a
Battelle technical staff member other than the person who originally generated the record.
Evaluation staff will be consulted as needed to clarify any issues 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.
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
Page 31 of 31
E REFERENCES
1. Quality Management Plan (QMP) for the Technology Testing and Evaluation Program
(TTEP), Version 1, prepared by Battelle, Columbus, Ohio, January 2005.
2. 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.
HealthRes., 11, 128-148(2001).
3. Decontamination Technology Testing and Evaluation: Task 1: Technology Identification
and Selection, prepared for the U.S. EPA under Task Order 1113 by Battelle, Columbus,
Ohio, December 17, 2004.
4. Battelle MREF SOP Number: MREF. X-074, "Standard Operating Procedure (SOP) for the
Production of Bacillus anthracis Spores."
5. Battelle MREF SOP Number: MREF. X-093, "Standard Operating Procedure (SOP) for the
Production of Bacillus anthracis Spores in a Small Fermentor."
6. Battelle MREF Facility Safety Plan Annex 12 to Appendix B, "Guidelines for the Use of
Class II and Class III Biological Safety Cabinets in the MREF Biofacility."
7. Battelle MREF SOP Number: MREF. X-054, "Standard Operating Procedure (SOP) for
Enumeration of BL-2 and BL-3 Bacterial Samples via the Spread Plate Technique."
8. Battelle MREF SOP Number: MREF. X-l 12, "Standard Operating Procedure (SOP) for
Interpreting and Calculating Enumeration Data for Biological Decontamination Testing."
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
APPENDIX
DETAILED DESCRIPTIONS OF DECONTAMINATION TECHNOLOGIES AND
OPERATING PARAMETERS FOR EFFICACY EVALUATION
Sabre Technical Services Chlorine Dioxide Fumigant Technology
General Description:
C1O2 is not stable as a compressed gas and, therefore, vaporous C1O2 must be produced on-site.
Decontamination technologies that utilize vaporous C1O2 typically include the equipment and
chemicals for on-site generation, delivery, removal and neutralization of C1O2. In addition, C1O2
generation technologies may require concurrent use of equipment for establishing and
maintaining specific temperature and humidity conditions that are required for safe and effective
operation of the decontamination process.
Sabre Equipment Description and Operating Parameters
The Sabre equipment includes a 20.3 cm x 20.3 cm (8" x 8") base onto which is mounted a 15.2
cm x 15.2 cm (6" x 6"), 91.4 cm (36") high sparging column. A 19 L (5 gallon) container
(vented through a sodium thiosulfate trap, container placed in an over pack for safety) containing
15 L of an aqueous solution consisting of 3g/L of CICh plus 1000 ppm of chlorite is prepared on-
site. The C1O2 solution is pumped (using a peristaltic pump) into the sparging column and air
from the test chamber is pumped into and through the column to sparge the C1O2 from the liquid
into the air stream; the air stream re-enters the glove box to establish the desired gaseous C1O2
concentration. Liquid introduction from the reservoir of CICVchlorite solution to the sparging
column is initially at the rate of 60 mL per minute; when the desired C1O2 concentration in the
test chamber is achieved, the liquid introduction into the sparging column is decreased to 3 mL
per minute. The spent liquid exiting the sparging column is collected in a reservoir. The air
from the chamber is recirculated into and out of the sparging column. Temperature for the
decontamination is held in the range of 23.9-35° C and the relative humidity should be held in
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
the range of 23.9-35° C. A nebulizer (supplied by Battelle for this test) is used to establish the
desired humidity level in the test chamber. A constant temperature bath is employed to maintain
the temperature of the liquid entering the sparging tower. The parameters of temperature and
humidity will not be varied in this evaluation. Total treatment time is 3 hours for 3,000 ppm
C1O2 in order to achieve a CT of 9,000.
At the end of the decontamination test the C1O2 in the system is removed by pumping through a
carbon adsorption column.
Description of the Method for Real-Time Monitoring of ClOi
The concentration of C1O2 inside the glove box will be monitored using a portable ultraviolet
(UV) spectrometer (developed by DARPA) for real-time monitoring of the concentration of C1O2
in air. The UV spectrometer measures the strong 360 nm UV absorption of C1O2 gas, generates
a 4-20 mA signal and, based on the absorption data, the ClO2gas concentration is calculated.
The UV optical beam is produced by a low power light emitting diode and is detected by a
photodiode. A drawing of the monitor assembly is shown in the Figure below (lower panel), and
a photograph of the as-built units is shown (upper picture).
-------
Sporicidal Decontamination Technologies Test/QA Plan
Date: 3/22/05
Version 1
The calibration of the UV spectrometer will be checked at the start of the study by comparing the
concentrations indicated by the UV spectrometer (concentrations in the range of 1000 ppmv to
3000 ppmv) by sampling the air in the glove box and comparing the C1O2 concentrations found
using OSHA Method ID 202 [see www.osha.gov/dts/sltc/methods/inorganic/id202/id202.html
(Note: This method was modified by reducing the volume of air captured to only 0.5 mL and
increasing the potassium iodide capture solution volumes to 50 mL)] with concentrations
determined using the DARPA UV spectrometer. The concentrations of C1O2 as determined
using the DARPA UV spectrometer will be deemed acceptable if the concentration determined
using the DARPA UV spectrometer (over the C1O2 concentrations of 1000 ppm to 3200 ppm)
agrees within ± 30% of the corresponding concentration determined using OSHA Method ID
202. [Note: Data indicative of the performance of OSHA Method ID 202 will be included in the
section of the report that describes the effort to calibrate the DARPA UV spectrometer.]
------- |