EPA/600/R-21/105 | August 2021
www.epa.gov/emergency-response-research
United States
Environmental Protection
Agency
&EPA
Remediation Options for
Fentanyl Contaminated
Indoor Environments
Office of Research and Development
Homeland Security Research Program
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Remediation Options for Fentanyl-
contaminated Indoor Environments
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EPA/600/R-21/105 | August 2021
Remediation Options for Fentanyl-contaminated Indoor Environments
U.S. Environmental Protection Agency
Office of Research and Development
Center for Environmental Solutions and Emergency Response
Research Triangle Park, NC 27711
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EPA/600/R-21/105 | August 2021
DISCLAIMER
The U.S. Environmental Protection Agency (EPA) through its Office of Research and
Development funded and managed the research described herein under Contract Number EP-C-
16-014, Task Order 68HERC20F0076 with Battelle. It has been subjected to the Agency's
review and has been approved for publication. Note that approval does not signify that the
contents necessarily reflect the views of the Agency. Any mention of trade names, products, or
services does not imply an endorsement by the U.S. Government or EPA. The EPA does not
endorse any commercial products, services, or enterprises. The contractor role did not include
establishing Agency policy.
Questions concerning this document, or its application should be addressed to:
Lukas Oudejans, Ph.D.
Homeland Security and Materials Management Division
Center for Environmental Solutions and Emergency Response
Office of Research and Development
U.S. Environmental Protection Agency (MD-E343-06)
109 T.W. Alexander Drive
Research Triangle Park, NC 27711
Phone: 919-541-2973
Fax: 919-541-0496
E-mail: Oudeians,Lukas@epa.gov
in
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EPA/600/R-21/105 | August 2021
FOREWORD
The U.S. Environmental Protection Agency (EPA) is charged by Congress with protecting the
Nation's land, air, and water resources. Under a mandate of national environmental laws, the
Agency strives to formulate and implement actions leading to a compatible balance between
human activities and the ability of natural systems to support and nurture life. To meet this
mandate, EPA's research program is providing data and technical support for solving
environmental problems today and building a science knowledge base necessary to manage our
ecological resources wisely, understand how pollutants affect our health, and prevent or reduce
environmental risks in the future.
The Center for Environmental Solutions and Emergency Response (CESER) within the Office of
Research and Development (ORD) conducts applied, stakeholder-driven research and provides
responsive technical support to help solve the Nation's environmental challenges. The Center's
research focuses on innovative approaches to address environmental challenges associated with
the built environment. We develop technologies and decision-support tools to help safeguard
public water systems and groundwater, guide sustainable materials management, remediate sites
from traditional contamination sources and emerging environmental stressors, and address
potential threats from terrorism and natural disasters. CESER collaborates with both public and
private sector partners to foster technologies that improve the effectiveness and reduce the cost
of compliance, while anticipating emerging problems. We provide technical support to EPA
regions and programs, states, tribal nations, and federal partners, and serve as the interagency
liaison for EPA in homeland security research and technology. The Center is a leader in
providing scientific solutions to protect human health and the environment.
This report assesses decontamination options for fentanyl contaminated building materials as
well as responder gear and personal protective equipment related materials. This report builds on
a previous fentanyl decontamination efficacy study that looked at other decontaminants. The
focus in this report is on decontamination products that were previously not tested and variations
in decontamination applications and their use to clean responder gear or personal protective
equipment (PPE).
Gregory Sales, Director
Center for Environmental Solutions and Emergency Response
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EPA/600/R-21/105 | August 2021
ACKNOWLEDGMENTS
This research is part of the U.S. Environmental Protection Agency's (EPA's) Homeland Security
Research Program's (HSRP) efforts to evaluate surface-applied liquid-based decontamination
methodologies for decontamination of fentanyl on building materials and responder gear or
personal protective equipment (PPE). Funding was provided through the regional applied
research effort (RARE) program, administrated by the Office of Science, Policy and Engagement
(OSAPE) under RARE Project 2082 entitled "Remediation of Fentanyl Contaminated Indoor
Environments".
This effort was directed by the principal investigator (PI) from the Office of Research and
Development's (ORD's) Homeland Security and Materials Management Division (HSMMD)
within the Center for Environmental Solutions and Emergency Response (CESER). The
contributions of the following individuals have been a valued asset throughout this effort.
EPA Project Team
Lukas Oudejans, ORD/CESER/HSMMD (PI)
Matthew Magnuson, ORD/CESER/HSMMD
James Justice, Region 5
Catherine Young, Region 1
Charlie Fitzsimmons, Region 3
Brian Englert, Region 4
Kerry Guy, Region 8
Battelle
David See (now Columbus Division of Fire)
Carissa Dodds
William Hayes
Melany Corlew
Anthony Ellingson
Jordan Vasko
Thomas Malloy
US EPA Technical Reviewers of Report
Paul Lemieux, ORD/CESER/HSMMD
John Archer ORD/CESER/HSMMD
US EPA Quality Assurance
Ramona Sherman, ORD/CESER/HSMMD
US EPA Technical Editing
Joan Bursey
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EPA/600/R-21/105 | August 2021
EXECUTIVE SUMMARY
The U.S. Environmental Protection Agency's (EPA's) Homeland Security Research Program
(HSRP) conducts research necessary for identification of methods and technologies that can be
used during hazardous materials remediation and cleanup efforts. The recent increase in the
number of unintentional fentanyl-related overdose fatalities in multiple states across the U.S. has
resulted in scenarios wherein local and state authorities request (technical) support from EPA in
the remediation of indoor fentanyl contamination at a home or other facility.
One of the main scientific gaps in the development of an adequate fentanyl contamination
remediation response is related to a lack of knowledge of effective decontamination technologies
and the conditions for their application for the degradation of fentanyl on a material or surface.
Another decontamination-related gap is associated with a lack of information on suitable
cleaning approaches for first responder gear or hazardous material responder personal protective
equipment (PPE) that may have become contaminated with fentanyl during the response or
remediation activities.
This project builds on an earlier fentanyl decontamination study which assessed efficacies of
several decontamination technologies given a single set of application conditions for the
degradation of fentanyl on the surface of selected commonly encountered indoor materials. The
purpose of this project was to evaluate the efficacy of two hydrogen peroxide-based
decontamination technologies that were either low cost, easy to acquire commercial off the shelf
(COTS) alternatives to specialized decontaminants, or technologies that were included as part of
a completed remediation of a fentanyl-contaminated property. For two other previously studied
decontaminants, namely, Dahlgren Decon™ and pH 5 adjusted bleach, the current study assessed
whether a reapplication of these decontaminants could improve overall efficacy, especially in the
presence of a benign additive that was demonstrated to create a demand on the decontamination
solution. Lastly, Dahlgren Decon™ and pH 5 adjusted bleach were also assessed on their ability
to degrade fentanyl on a short dwell timescale of only a few minutes. Such a short dwell time
would occur if these decontaminants were applied as part of a decontamination line procedure in
which the PPE or other response gear is cleaned to reduce exposure to support personnel and
eliminate spread of contamination when exiting a (fentanyl) contaminated site.
Decontaminants that were considered included: Meth Remover® and ZEP® Professional Stain
Remover with Peroxide (both hydrogen peroxide-based), Dahlgren Decon™ (activated peracetic
acid as active ingredient), and pH 5 adjusted bleach with surfactant (hypochlorite based) derived
from Clorox™ ProResults® Garage and Driveway Cleaner (referred to as pH 5 modified
surfactant bleach). Four indoor-related materials were selected for this study: painted drywall,
laminate, powder-coated steel, and wood. First responder PPE materials included neoprene,
Saranex®, DuraChem® 500 Level B HazMat suit, and bunker gear. Fentanyl hydrochloride
(HC1; 1 mg) was applied as a solid powder to the surface of replicate materials with 10-cm2
surface area.
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EPA/600/R-21/105 | August 2021
Decontaminants were applied via a spray at a target application volume of 60 |iL/cm2. Following
the specific decontaminant dwell period, coupons and decontaminant runoff were extracted with
an organic solvent and extracts were analyzed via gas chromatography/mass spectrometry
(GC/MS) or liquid chromatography-tandem mass spectrometry (LC-MS/MS) to quantify the
amount of fentanyl HC1 remaining in the extracts.
Results
• Measured efficacies for both hydrogen peroxide-based decontaminants were noticeably
modest and ranged from 14% to 46% (ZEP® product) and 23% to 58% for Meth
Remover® across the four materials. We can conclude that hydrogen peroxide is not a
highly effective degradant for fentanyl.
• The reapplication of Dahlgren Decon™ and pH 5 modified surfactant bleach did not
significantly improve efficacy versus the single application investigated as part of the
previous fentanyl decontamination study. Here, Dahlgren Decon™ yielded a greater than
99.8%) decontamination efficacy across all materials while the pH 5 modified surfactant
bleach yielded 80% - 96% efficacy, depending on the material. A direct comparison
between the previous and current study was complicated by the differences in materials
and recoveries from positive controls.
• Both Dahlgren Decon™ and pH 5 modified surfactant bleach achieved lower efficacies
for decontamination of fentanyl mixed with ascorbic acid as a challenging benign
additive, namely 97% (99.8% without additive) on wood for Dahlgren Decon™ and 80%
(84%) without additive) on wood for pH 5 modified surfactant bleach.
• A diluted (1:4) Dahlgren Decon™ and pH 5 modified surfactant bleach solutions were
able to degrade fentanyl over a short (5-minute), duration with efficacies ranging from
89%) - 98%) across materials for diluted Dahlgren Decon™ and 55% - 66% for the pH 5
modified surfactant bleach.
Figure ES-1 summarizes the average percent decontamination efficacies measured for each test
condition.
In many decontamination tests, the observed large variation in amounts recovered can be linked
to the presence of agglomerated fentanyl on the surface during the application of the
decontaminant spray, resulting in a slower mass transfer rate between decontaminant and
fentanyl and hence, higher amounts recovered even in the presence of an otherwise effective
decontaminant.
The results of this work inform EPA responders, governments, and health departments in their
guidance development for decontamination technology recommendations for building materials
contaminated with fentanyl.
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EPA/600/R-21/105 | August 2021
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TABLE OF CONTENTS
DISCLAIMER Ill
FOREWORD IV
ACKNOWLEDGMENTS V
EXECUTIVE SUMMARY VI
TABLE OF CONTENTS IX
LIST OF TABLES XI
LIST OF FIGURES XII
LIST OF ACRONYMS AND ABBREVIATIONS XIII
INTRODUCTION 1
1.1 Purpose 1
1.2 Project Objectives 1
1.3 Test Facility Description 2
1.4 Staff and Resources 2
EXPERIMENTAL METHODS 3
2.1 Experimental Design 3
2.1.1 Test Methods 3
2.1.2 Decontamination Efficacy Evaluation 8
2.2 Experimental Methods and Materials 11
2.2.1 Coupon Materials 11
2.2.2 Fentanyl 13
2.2.3 Application of Decontamination Technologies 15
2.2.4 Extraction of Fentanyl from Coupons 21
2.3 Analytical Methods 21
2.3.1 Quantitative Fentanyl Analysis - GC/MS 22
2.3.2 Quantitative Fentanyl Analysis - LC-MS/MS 22
2.4 Calculations 24
2.4.1 Decontamination Efficacy Evaluation 24
2.4.2 Decontamination Efficacy Evaluation (Benign Additive Ascorbic Acid) 26
2.5 Statistical Analyses 26
2.5.1 Group 1: Comparisons of Decontaminant Performance within Material Type 27
2.5.2 Group 2: Decontamination Period Effect Analysis Plan 28
2.5.3 Group 3: Challenge Additive Effect Analysis Plan 28
2.5.4 Group 4: Decontaminant Effect Analysis Plan 28
RESULTS 30
3.1 Methods Demonstration 30
3.1.1 Fentanyl Delivery (Spiking) Characterization 30
3.1.2 Decontaminant Spray Delivery Characterization 30
3.1.3 Quench Method Demonstration 31
3.2 Decontamination Efficacy Evaluation - Building Materials 33
3.2.1 Hydrogen Peroxide-Based Decontaminants 33
3.2.2 Decontamination Efficacy Evaluation - Reapplication of Decontaminants 35
3.2.3 Decontamination Efficacy Evaluation - PPE/Gear Materials 38
3.3 ANOVA Results 41
3.3.1 Positive Control Comparison Results 41
3.3.2 Comparison of Test Sample Results 46
3.3.3 Limitations of Statistical Analysis 52
QUALITY ASSURANCE/QUALITY CONTROL 53
IX
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4.1 Data Quality Indicators 53
4.2 Instrument Calibration 54
4.2.1 Calibration Schedules 54
4.2.2 LC-MS/MS Calibration 55
4.2.3 GC/MS Calibration 57
4.3 Sample Handling and Custody 58
4.4 Test Parameter Control Sheets 58
4.5 Technical Systems Audit 58
4.6 Performance Evaluation Audits 58
4.7 Data Quality Audit 59
4.8 Q APP Amendments 60
4.9 QAPP Deviations 60
DISCUSSION/CONCLUSIONS 61
5.1 Building Material Decontamination 61
5.2 PPE/Responder Gear Decontamination 62
5.3 Clustering of Fentanyl Powder 62
REFERENCES 63
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LIST OF TABLES
Table 1. Quench Method Scoping Test Matrix 6
Table 2. Decontamination Efficacy Test Matrix - Hydrogen Peroxide Solutions 9
Table 3. Decontamination Efficacy Test Matrix 10
Table 4. Decontamination Efficacy Test Matrix for PPE Materials 10
Table 5. Coupon Materials 13
Table 6. GC/MS Conditions for Quantitative Fentanyl Analysis 22
Table 7. Analyte Ion Transitions 23
Table 8. LC-MS/MS Conditions for Quantitative Fentanyl Analysis 23
Table 9. Decontaminant Spray Application Settings 31
Table 10. Decontaminant Spray Delivery Summary 31
Table 11. Quench Method Demonstration Test, Average Mass Recovery 32
Table 12. Decontamination Efficacy Testing, Spike Control Average Recovery 33
Table 13. Decontamination Efficacy Testing, Spike Control Average Recovery 36
Table 14. Average Mass Recovery, Diluted Dahlgren Decon™ 39
Table 15. Decontamination Efficacy Testing, Spike Control Average Recovery 39
Table 16. ANOVA Results for Dahlgren Decon™ at 60 + 60 min (Positive Controls) 43
Table 17. ANOVA Results for Diluted Dahlgren Decon™ at 5 min (Positive Controls) 43
Table 18. ANOVA Results for pH 5 Modified Surfactant Bleach at 60 + 60 min (Positive Controls) 43
Table 19. ANOVA Results for pH 5 Modified Surfactant Bleach at 5 min (Positive Controls) 43
Table 20. ANOVA Results for Dahlgren Decon™, Collapsed over Multiple Materials (Positive Controls) 44
Table 21. ANOVA Results for pH 5 Modified Surfactant Bleach, Collapsed over Multiple Materials (Positive
Controls) 44
Table 22. ANOVA Results for Dahlgren Decon™ on Wood at 60 + 60 min (Positive Controls) 44
Table 23. ANOVA Results for pH 5 Modified Surfactant Bleach on Wood at 60 + 60 min (Positive Controls) 44
Table 24. ANOVA Results on Laminate at 60 min (Positive Controls) 45
Table 25. ANOVA Results for Dahlgren Decon™ at 60 + 60 min (Test Samples) 48
Table 26. ANOVA Results for Diluted Dahlgren Decon™ at 5 min (Test Samples) 48
Table 27. ANOVA Results for pH 5 Modified Surfactant Bleach at 60 + 60 min (Test Samples) 48
Table 28. ANOVA Results for pH 5 Modified Surfactant Bleach at 5 min (Test Samples) 49
Table 29. ANOVA Results for Dahlgren Decon™, Collapsed over Multiple Materials (Test Samples) 49
Table 30. ANOVA Results for pH 5 Modified Surfactant Bleach, Collapsed over Multiple Materials (Test Samples)
49
Table 31. ANOVA Results for Dahlgren Decon™ on Wood at 60 + 60 min (Test Samples) 50
Table 32. ANOVA Results for pH 5 Modified Surfactant Bleach on Wood at 60 + 60 min (Test Samples) 50
Table 3 3. ANOVA Results on Laminate at 60 min (Test Samples) 51
Table 34. Data Quality Indicators and Results 53
Table 35. Equipment Calibration Schedule 55
Table 36. LC-MS/MS Analysis Performance Parameters and Acceptance Criteria 57
Table 37. Performance Evaluation Audit Results 59
Table Bl. Environmental Conditions in Experimental Chamber with Spray Setup 71
Table CI. Decontaminant Spray Delivery Mass per Position, Meth Remover® 73
Table C2. Decontaminant Spray Delivery Mass per Position, ZEP® 73
Table C3. Decontaminant Spray Delivery Mass per Position, pH 5 Modified Surfactant Bleach 74
Table C4. Decontaminant Spray Delivery Mass per Position, Dahlgren Decon™ 74
Table C5. Decontaminant Spray Delivery Mass per Position, Diluted Dahlgren Decon™ 75
Table Dl. Average Mass Recovery, Meth Remover® 77
Table D2. Average Mass Recovery, ZEP® 77
Table D3. Decontamination Efficacy Testing, Average Percent Efficacy 78
Table D4. Average Mass Recovery, Dahlgren Decon™ - Double Application 78
Table D5. Average Mass Recovery, pH 5 Modified Surfactant Bleach - Double Application 79
Table D6. Decontamination Efficacy Testing, Average Percent Efficacy 79
Table D7. Average Mass Recovery, Dahlgren Decon™ 80
Table D8. Average Mass Recovery, pH 5 Modified Surfactant Bleach 80
Table D9. Decontamination Efficacy Testing, Average Percent Efficacy 81
XI
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LIST OF FIGURES
Figure ES-1. Average decontamination efficacies viii
Figure 1. Drummond pipettor for delivery of solids 4
Figure 2. Handheld pump pressurization-style sprayer 17
Figure 3. Sprayer and test chamber setup 18
Figure 4. Decontaminant spray tray setup 19
Figure 5. Spray application 20
Figure 6. Test sample arrangement under sprayer 30
Figure 7. Quench method demonstration test, average percent recovery by GC/MS 32
Figure 8. Quench method demonstration test, average percent recovery by LC-MS/MS 32
Figure 9. Decontamination efficacy testing, average mass recovery 34
Figure 10. Decontamination efficacy testing, average percent efficacy 35
Figure 11. Decontamination efficacy testing, average mass recovery 37
Figure 12. Decontamination efficacy testing, average percent efficacy 38
Figure 13. Decontamination efficacy testing of PPE materials, average mass recovery 40
Figure 14. Decontamination efficacy testing, average percent efficacy 41
ATTACHMENTS
Attachment A - Fentanyl Certificate of Analysis
Attachment B - Environmental Data
Attachment C - Spray Characterization Data
Attachment D - Average Mass Recovery and Decontamination Efficacy Data
xii
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LIST OF ACRONYMS AND ABBREVIATIONS
ANOVA
analysis of variance
°C
degrees Celsius
CAS
Chemical Abstracts Service
CBRN
Chemical, Biological, Radiological, Nuclear
CCV
continuing calibration verification
CESER
Center Environmental Solutions and Emergency Response (U.S. EPA)
cm
centimeter(s)
cm2
square centimeter(s)
CoC
chain of custody
COTS
commercial off the shelf
CoV
coefficient of variation
DEA
U.S. Drug Enforcement Agency
DFTPP
decafluorotriphenylphosphine
EPA
U.S. Environmental Protection Agency
FEP
fluorinated ethylene propylene
g
gram(s)
GC
gas chromatography
GC/MS
gas chromatography/mass spectrometry
h
hour(s)
HC1
hydrochloride
HMRC
Hazardous Materials Research Center
HPLC
high performance liquid chromatography
HSMMD
Homeland Security and Materials Management Division (U.S. EPA)
HSRP
Homeland Security Research Program (U.S. EPA)
in2
square inch(es)
IPA
isopropyl alcohol
IS
internal standard
LC-MS/MS
liquid chromatography-tandem mass spectrometry
LLOQ
lower limit of quantitation
LRB
laboratory record book
M
Molar
Mg
microgram(s)
jiL
microliter(s)
mg
milligram(s)
min
minute(s)
mL
milliliter(s)
mm
millimeter(s)
MRM
multiple reaction monitoring
MSD
mass selective detector
NFPA
National Fire Protection Agency
ng
nanogram(s)
NIST
National Institute of Standards and Technology
ORD
Office of Research and Development (U.S. EPA)
OSAPE
Office of Science, Policy and Engagement (U.S. EPA)
PE
performance evaluation
PFPP
pentafluorophenylpropyl
PI
principal investigator
PP
polypropylene
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PPE
personal protective equipment
psi
pound(s) per square inch
PTFE
polytetrafluoroethylene
PVC
polyvinyl chloride
QA
quality assurance
QAPP
quality assurance project plan
QC
quality control
r2
coefficient of determination
RARE
Regional Applied Research Effort
RH
relative humidity
RSD
relative standard deviation
RT
retention time
RTU
ready to use
SC
spike control
sec
seconds
SIM
selected ion monitoring
STS
sodium thiosulfate (Na2S:Ch)
TPCS
test parameter control sheet
TSA
technical systems audit
V
Volt
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INTRODUCTION
The U.S. Environmental Protection Agency (U.S. EPA) is responsible for preparing for,
responding to, and recovering from threats to public health, welfare, or the environment caused
by actual or potential hazardous materials incidents. Hazardous materials include chemical,
biological, and radiological substances, whether accidentally or intentionally released. The
imminent threat of a chemical agent release into the environment is driving EPA's Homeland
Security Research Program (HSRP) to systematically evaluate potential decontamination
technologies for chemical agents.
Fentanyl is a synthetic, short-acting opioid analgesic that is 50-100 times more potent than
morphine. Fentanyl has been approved for managing acute or chronic pain associated with
advanced cancer. Although pharmaceutical fentanyl can be diverted for misuse, most cases of
fentanyl-related morbidity and mortality have been linked to illicitly imported or manufactured
fentanyl that is sold via illicit drug markets. Illicit-use fentanyl is often mixed with heroin,
cocaine, or more benign additives. The recent increase in unintentional fentanyl-related overdose
fatalities in multiple states across the U.S. has resulted in scenarios where local and state
authorities request (technical) support from EPA in the remediation of indoor fentanyl
contamination at a residence or other type of facility. One of the main scientific gaps in
development of an adequate remediation response is related to a lack of knowledge of effective
decontamination technologies and their application conditions for degradation of fentanyl on a
material or surface. Current remediation efforts tend to rely on physical removal approaches
including careful dry vacuuming followed by "soap and water" cleaning.
A first systematic fentanyl decontamination study has recently been completed [1], That study
assessed efficacy of multiple decontamination solutions including water (reference solution),
OxiClean™, bleach, pH adjusted bleach (both at pH 7 and 5), EasyDecon DF200, and Dahlgren
Decon™. All tests were conducted with fentanyl applied to stainless steel, laminate, acrylic and
painted drywall as a dry powder. Decontaminants were applied via spray for a fixed 1-hour (h)
dwell period.
The fentanyl decontamination tests described in this report build on the knowledge gained from
this previous study through the addition of decontamination solutions, changes in application
procedures as well as an investigation into the ability of two of the better performing
decontamination solutions to degrade fentanyl on personal protective equipment (PPE)-related
materials over a short (5-minute [min]) dwell time.
1.1 Purpose
The purpose of this project was to evaluate the efficacy of various decontamination technologies
to degrade fentanyl on the surface of commonly encountered building- or PPE-related materials.
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1.2 Project Objectives
Specific objectives of this study included:
• Assessment of hydrogen peroxide-based commercially available decontamination
technologies that are anticipated to be efficacious in degrading fentanyl contamination on
hard nonporous material surfaces.
• Determine change in overall efficacy after reapplication of technologies that
demonstrated the highest degree of efficacy during the previous decontamination tests
and materials commonly found in indoor settings [1],
• Evaluating decontamination efficacy through additional testing with a significantly
shorter dwell time of 5 min for responder gear or PPE materials.
1.3 Test Facility Description
All tests were performed at Battelle's Hazardous Materials Research Center (HMRC) located in
West Jefferson, Ohio. The HMRC is registered with both the U.S. Drug Enforcement Agency
(DEA) and the Ohio Board of Pharmacy to procure, store, synthesize, and use controlled
substances up to and including DEA Schedule I material. Wherever applicable and required, the
reporting requirements of these registrations were followed.
1.4 Staff and Resources
Surface decontamination efficacy testing and all associated method development testing and
sample analyses were completed using staff and resources from Battelle's HMRC in consultation
with the EPA Center for Environmental Solutions and Emergency Response (CESER) Homeland
Security and Material Management Division (HSMMD).
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EXPERIMENTAL METHODS
2.1 Experimental Design
Decontamination efficacy was evaluated through execution of surface decontamination tests.
Research was limited to decontamination of fentanyl hydrochloride (HC1) and did not include
analogs (e.g., carfentanil). Four (4) commercially available decontaminants were tested (refer to
Section 2.1.2) to evaluate the efficacies of the technologies in the degradation of solid fentanyl
HC1 (CAS 1443-54-5, from here on referred to as fentanyl unless otherwise specified) from the
surface of four (4) commonly encountered, indoor-related building materials as well as from the
surface of four (4) responder gear or PPE materials. Prior to the surface decontamination efficacy
tests, the test methods were experimentally demonstrated, and results were evaluated against
predefined criteria to ensure valid data would be generated.
Individual test articles consisted of small coupons of the indoor building or gear-/PPE-related
materials. Coupons measured 2.5 centimeters (cm) x 4 cm (10 square centimeters [cm2]
contamination/decontamination surface area). Coupon thickness was dependent on the specific
material type. Refer to Section 2.2.1 for more information on the test articles and indoor and
PPE-related materials used during the evaluation.
During all decontamination testing, decontaminants were delivered by a sprayer system that
produced a low-pressure spray similar to common commercially available, backpack-type
sprayers and were applied via spray directly onto the surface of the test articles.
Following decontamination (dwell time varied based on purpose of the test), test articles were
extracted with a solvent and the extract as well as decontamination rinsate were analyzed by gas
chromatography/mass spectrometry (GC/MS) or liquid chromatography/tandem mass
spectrometry (LC-MS/MS) to quantify the total amount of residual fentanyl (freebase and salt).
Analysis via GC/MS was anticipated for samples with fentanyl concentrations in the range of
350 micrograms ([^/milliliter (mL) down to 0.05 |ig/mL, LC-MS/MS was used for analysis of
samples with fentanyl concentrations that fall within the range of 5 nanograms (ng)/mL down to
0.01 ng/mL.
2.1.1 Test Methods
The test methods used were experimentally verified/demonstrated, and results were evaluated
against predefined criteria to ensure valid data would be generated.
2.1.1.1 Fentanyl Delivery (Spiking) Characterization
Solid fentanyl was applied to test articles using a 50-|iL Drummond Series 500 Digital
microdispenser (3-000-550, Drummond, Broomall, PA) (Figure 1).
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M I
Figure 1. Drummond pipettor for delivery of solids.
During use, the capillary was pressed into the solid fentanyl at least three times to ensure
adequate packing of material into the capillary underneath the plunger. The solid agent was then
dispensed onto the test article surface. Refer to Section 2.2.2.2 for additional information
regarding application of fentanyl to coupons. The mass application target was 1 milligram (mg),
which required a Drummond setting of 1.9 microliters (|iL) based on previous use to deliver
fentanyl.
The adequacy of the Drummond to deliver accurate and reproducible amounts of solid fentanyl
onto the surfaces of coupons was also continuously evaluated during execution of the method
demonstration tests (Sections 2.1.1.4) and decontamination efficacy tests (Section 2.1.2). As
described for the approaches for these test phases, fentanyl spike controls were generated during
tests by delivering the same mass of fentanyl as the mass applied to test coupons into extraction
jars, dissolving the fentanyl in extraction solvent, and analyzing the spike control extracts
alongside the test and control samples. A comparison between the spike control sample results
and the theoretical target fentanyl delivery mass provided information on the accuracy and
reproducibility of the Drummond method to deliver solid fentanyl onto coupons. The target spike
control recovery criteria were set at 80% to 120% of the theoretical mass with less than 30%
relative standard deviation [RSD], see Table 34 in Section 4.1. The spike control results are
provided together with the associated test sample results in Sections 3.1.3 and 3.2.
During the previous study, it was postulated that the Drummond capillary was repeatedly pressed
into the solid fentanyl in the working vial leaving void spaces in the powder after the Drummond
capillary was withdrawn. These void spaces led to inconsistent Drummond capillary loading. To
eliminate the void spaces, regular stirring of the fentanyl powder in the working vial using the
Drummond capillary was implemented. Specifically, a stainless steel microspatula was used to
stir and mix the powder prior to each fentanyl spiking operation and then again during the
operation (after approximately half of the samples of the test had been spiked). Decreased
variability (that is, variability within specification, i.e., < 30% RSD, refer to Section 4.1) across
spike control replicates included in subsequent tests was then achieved following implementation
of this regular stirring and mixing of the fentanyl within the vial to promote uniformity of the
powder.
Fentanyl was applied to the 10-cm2 coupons as a single (target) 1-mg pile placed in the center of
the coupon. Following application of fentanyl onto the surface of test and control coupons, the
solid was spread evenly across the test article using an antistatic spatula (14-245-99, Fisher
Scientific, Pittsburgh, PA). Fentanyl was spread evenly (subjective, visually) across
approximately 50% of the 10-cm2 area of the coupons. As part of the previous study, we had
4
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determined that the amount of fentanyl typically lost to the spatula was minimal (average of 9%
or less of the amount spiked), so the spatulas were not extracted during decontamination testing.
A different spatula was used for each test/control coupon subsection (i.e., spatulas were not
reused for multiple samples).
2.1.1.2 Solvent Extraction of Fentanyl from Coupons
The methods developed for solvent extraction of fentanyl from similar materials during previous
work [1] were not evaluated for effectiveness in recovery of fentanyl from the new materials that
were introduced in this study. Fentanyl-HCl as a powder binds loosely to nonporous surfaces and
can be extracted without difficulty. Confirmation of the high recoveries can be found in the
comparison of fentanyl recoveries from spike controls and associated positive controls for each
decontamination test. Based on solvent extraction recovery means and coefficient of variance
(CoVs) for each solvent evaluated during the previous fentanyl decontamination study (highest
recovery with lowest CoV), isopropyl alcohol (IPA) was selected for use during all phases of
testing.
2.1.1.3 Decontaminant Delivery Characterization
Liquid decontaminants were applied to test and control sample coupons via moderately low flow
spray using a nozzle typical of a pump pressurization style sprayer (12U469, Grainger, Lake
Forest, IL). Section 2.2.3.5 provides information on the sprayer that was used and how the
sprayer was interfaced with the test chamber and operated to deliver decontaminants to coupons
via uniform spray at the target application volume.
Prior to testing, operation of the sprayer was characterized using each decontamination
technology to determine the sprayer settings and use procedures (e.g., sprayer nozzle stand-off
distance, sprayer pressurization, spray sweep speed, etc.) necessary to deliver a target
decontaminant volume per unit area of 60 |iL/cm2. Additionally, spray settings were determined
such that in addition to delivering 60 |iL/cm2 of decontaminant to the surface of coupons, spray
impact pressure was minimized to reduce, to the greatest extent possible, physical removal of
spiked fentanyl (powder) from the surface of coupons (so that quantification of residual fentanyl
post decontamination could be attributed to chemical degradation rather than to physical
removal).
2.1.1.4 Decontamination Technology Quench and Matrix Effect Evaluation
During decontamination efficacy tests, residual decontaminant on the materials or in the runoffs
could be collected in the sample extracts and continue to decontaminate fentanyl. Additionally,
chemical compounds extracted from the indoor materials, residual decontaminants, and/or
quench agents could create complex sample matrices, which could lead to false-positive or false-
negative results and/or analytical interferences. Effective decontaminant quench methods were
necessary to allow measured decontamination efficacies to be associated with specific
5
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decontaminant dwell times. Similarly, assessment of matrix effects was also necessary to ensure
the matrices would not interfere with analyses.
During the previous fentanyl decontamination effort [1], two approaches were considered to
quench the reaction of the decontaminants with fentanyl. Dilution in extraction solvent alone was
first evaluated as a method for quenching reaction of the decontaminants with fentanyl. The
hypothesis was that dilution in excess extraction solvent would slow the decontamination
reaction enough to allow for measurement of efficacy after a defined period. Results of the
quench method test suggested that dilution in extraction solvent was ineffective in preventing
decontamination of the post-spiked fentanyl by Dahlgren Decon™ and pH 7 bleach. In parallel,
an alternative quench method was identified by the addition of 5 mL of a 3-molar (M) solution of
sodium thiosulfate (Na2S2C>3; STS) in water to the IPA used to extract coupons and runoff boxes.
This approach was demonstrated to effectively quench the decontamination of fentanyl by
Dahlgren Decon™ and pH 7 bleach. In this study, a procedure and single-test matrix was
developed to demonstrate the adequacy of 3M STS as a quench agent for halting
decontamination of fentanyl by the hydrogen peroxide decontaminants (Meth Remover® and
ZEP® Professional Stain Remover with Peroxide; hereafter ZEP®). Meth Remover® was
selected based on its use during the 2017 remediation of a fentanyl contaminated house [2],
ZEP® was selected as a low-cost hydrogen peroxide-based decontaminant with a relatively high
(approximately 4% v/v) hydrogen peroxide concentration and similar to the EasyDecon DF200
decontamination product that was evaluated during the previous study [1], Table 1 provides the
experimental matrix that was intended to serve two purposes: to evaluate 3M STS as an
appropriate quench method for quenching the residual fentanyl decontamination reactions of
Meth Remover® and Zep® Professional Stain Remover with hydrogen peroxide, and to evaluate
any effects of sample matrices due to residual decontaminant and/or the 3M STS quench agent
itself on analysis of fentanyl and the response of the fentanyl-d5 internal standard (IS).
Table 1. Quench Method Scoping Test Matrix
Decontaminant
Fentanyl
Target
Concentration
Description
Quench
Analysis
Meth Remover®
2 iift/mL
Quench Samples
3M STS
GC/MS
ZEP®
2 iift/mL
Quench Samples
3M STS
GC/MS
None
2 iift/mL
Spike Controls
NA
GC/MS
Meth Remover®
2 ng/mL
Quench Samples
3M STS
LC-MS/MS
ZEP®
2 ng/mL
Quench Samples
3M STS
LC-MS/MS
None
2 ng/mL
Spike Controls
NA
LC-MS/MS
Each decontaminant/quench combination described in Table 1 was tested in triplicate. Spike
controls were generated throughout the test (i.e., one spike control prior to delivery of fentanyl to
the quench samples, one in the middle of the operation, and one after all quench samples had
been spiked).
Sample extract matrices representative of decontaminant runoff test samples (regarding the
amount of decontaminant anticipated to be present in the runoff extracts following
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decontaminant application via spray onto coupons) were also prepared. During the previous
study, the volumes of decontaminant remaining on the surface of coupons and collected in the
decontaminant runoff following spray-application were characterized using water. The average
volume of water/decontaminant remaining on the surface of coupons following spray-application
was measured to be 0.21 mL, and the average volume of water/decontaminant collected in the
runoff was 0.99 mL. Thus, representative decontaminant runoff sample extracts were prepared
by adding 0.99 mL of test decontaminant to 20 mL of IP A with 5 mL of 3M STS (i.e., no indoor-
or PPE-related material coupons or spray application of decontaminants were used).
Representative extracts were thoroughly mixed via vortex following preparation. Representative
decontaminant runoff extract samples were prepared and used for quench method demonstration
testing since the runoff decontaminant volume is greater than the coupon decontaminant volume
(0.99 mL versus 0.21 mL) and provides the most technically conservative "worst case" quench
test scenario (since the same volume of 3M STS quench [5 mL] was used to quench residual
decontaminant in both coupon and runoff extracts).
Following preparation of the representative decontaminant runoff extracts, a dilute fentanyl
solution was post-spiked into the IPA layer of the extracts. Extracts intended for GC/MS analysis
were spiked with 40 [xL of a 1 mg/mL fentanyl solution to target a final concentration of 2
[j,g/mL in the extracts. Extracts intended for LC-MS/MS analysis were spiked with 40 |iL of a 1
[j,g/mL fentanyl solution to target a final concentration of 2 ng/mL in the extracts. Extracts were
mixed thoroughly again via vortex post-spike. Aliquots of the extracts were collected from the
IPA (top) layer and stored at -20 ± 10 degrees Celsius (°C) for 72 h. Following the 72-h period,
the extract aliquots were equilibrated to room temperature and analyzed via GC/MS or LC-
MS/MS (depending on post-spike concentration) to quantify fentanyl, and results were compared
to the anticipated concentrations (based on the post-spiked mass and assuming no
decontamination occurred) and appropriate control samples containing no decontaminant to
determine the effectiveness of the quench methods.
Addition of 5 mL of 3M STS to the IPA used to extract coupons (10 mL) and runoff samples (20
mL) during decontamination efficacy tests were considered sufficient to preserve residual
fentanyl in the extracts (i.e., prevent further decontamination of fentanyl past the tested
decontaminant dwell period) during storage for up to 72 h at -20 ± 10°C if the amounts of
fentanyl recovered from post-spiked extracts containing decontaminants (quench samples; three
(3) replicates per decontaminant/fentanyl post-spike concentration/analysis method combination)
were each at least 70% of the mean amount of fentanyl recovered from post-spiked extracts that
did not contain decontaminants (positive controls; three (3) replicates per fentanyl post-
spike/analysis method combination). The impact of extract matrix effects due to residual
decontaminants and/or the 3M STS quench on the accuracy of quantitative analyses was
considered negligible as well if both of the following criteria were met:
• Recovery of fentanyl > 70% of the theoretical post-spiked amount in representative
decontaminant runoff extract matrix samples (quench samples).
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• The quality assurance/quality control (QA/QC) criteria for fentanyl-d5 IS response
discussed in Sections 4.2.2 and 4.2.3 were satisfied.
If the above criteria were satisfied, we concluded that any matrix effects observed were
influencing response of both fentanyl and the fentanyl-d5 IS in an identical manner, and that the
IS was adequately and appropriately compensating for any effects and facilitating accurate
fentanyl quantitation. As described in Sections 4.2.2 and 4.2.3, IS was added to analytical
samples just prior to analysis to lessen the concern for decontamination/degradation of the IS in
the samples due to unquenched decontaminant.
2.1.2 Decontamination Efficacy Evaluation
A post-test only control group experimental design was used for the decontamination efficacy
evaluation. Decontamination was the experimental variable. Test coupons were contaminated,
decontaminated, sampled, and analyzed for fentanyl. Positive control coupons were
contaminated but not decontaminated and subsequently sampled and analyzed for fentanyl along
with the test coupons. The effect of decontamination (efficacy) was defined as the percentage of
fentanyl remaining (total residual active fentanyl, i.e., salt and freebase) on the test coupons
compared to the positive control coupons. The higher the efficacy, the greater the effect of
decontamination by the specific technology.
Procedurally, a target 1 mg of solid fentanyl was spiked and distributed onto the center portion of
each test (3 replicates) and positive control (3 replicates) material coupon as described in Section
2.2.2.2. Material coupons measured 4.0 cm long by 2.5 cm wide (10-cm2 coupon surface area).
The spiked coupons were allowed to remain undisturbed during a set fentanyl contact period of
60 min. Following the fentanyl contact period, the decontamination technology under test was
applied as a liquid spray directly to the fentanyl challenge on each test coupon and allowed to
remain in contact with the fentanyl for the targeted dwell time.
Decontamination technology application procedures are described in Section 2.2.3.5. Following
the decontamination period, the test and positive control coupons were sampled for residual
fentanyl via solvent extraction according to Section 2.1.1.4 using 10 mL of IPA with 5 mL of 3M
STS quench. Coupon extracts were then analyzed for residual fentanyl via GC/MS or LC-
MS/MS according to Section 2.3.1 and 2.3.2.
During application of the decontaminants via moderately low flow spray, we anticipated that a
portion of the (target) 600 |iL delivered over each coupon would run off the coupon surface.
Coupons were placed in individual acrylic boxes on top of polypropylene (PP) mesh disks to
allow for collection of the runoff while elevating the coupons out of the decontaminant liquid
that was collected (refer to Section 2.2.3.6). Decontaminant runoff from each coupon was
collected, and each runoff sample was analyzed via GC/MS or LC-MS/MS to quantify any
residual fentanyl. Refer to Sections 2.2.3.5 and 2.2.3.6 for details related to spray application of
decontaminants and collection of the associated decontaminant runoff from each coupon. Runoff
analysis results provided indication of physical removal of fentanyl from the coupon surface.
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The matrix for decontamination efficacy testing for the two hydrogen-peroxide-based
decontaminants is provided in Table 2. During each test, environmental conditions (temperature
and relative humidity [RH]) were monitored and recorded, but not controlled.
Table 2. Decontamination Efficacy Test Matrix - Hydrogen Peroxide Solutions
Test
No.
Sample Type
Material
Fentanyl
Challenge
Decontamination
Technology
Dwell Time*
(min)
Replicates
Test Sample
Painted drywall
1 mg
Meth Remover®
60
3
Positive Control
Painted drywall
1 mg
None
60
3
Test Sample
Laminate
1 mg
Meth Remover®
60
3
1
Positive Control
Laminate
1 mg
None
60
3
Test Sample
Coated steel
1 mg
Meth Remover®
60
3
Positive Control
Coated steel
1 mg
None
60
3
Test Sample
Wood
1 mg
Meth Remover®
60
3
Positive Control
Wood
1 mg
None
60
3
Test Sample
Painted drywall
1 mg
Zep®
60
3
Positive Control
Painted drywall
1 mg
None
60
3
Test Sample
Laminate
1 mg
Zep®
60
3
2
Positive Control
Laminate
1 mg
None
60
3
Test Sample
Coated steel
1 mg
Zep®
60
3
Positive Control
Coated steel
1 mg
None
60
3
Test Sample
Wood
1 mg
Zep®
60
3
Positive Control
Wood
1 mg
None
60
3
In the previous fentanyl decontamination study [1], high efficacies (better than 99%) were
measured for the peracetic acid-based Dahlgren Decon™ product as well as the hypochlorite-
based chemistry in pH 5 modified surfactant bleach. Here, these two decontaminants were
applied as a liquid spray directly to the fentanyl challenge on each test coupon and allowed to
remain in contact with the fentanyl for 60 min. At that time, coupons were tilted to allow residual
decontaminant to run off the surface into the collection box. This process was followed by a
second application of the decontaminant as a liquid spray, which was allowed to remain in
contact with the fentanyl on the surface for another 60 min. The matrix for decontamination
efficacy testing for these two decontaminants is provided in Table 3.
As a continuation and natural progression of the decontamination efficacy evaluations, additional
decontamination efficacy tests were conducted that focused on evaluation of the efficacy of
selected decontaminants for degradation of a formulation of fentanyl directly on the surface of
indoor-related materials. As it pertains to this testing, a formulation of fentanyl was defined as
fentanyl mixed with a benign additive as may be encountered in samples collected in the field. In
this study, ascorbic acid (Vitamin C; PHR1008-2G, Millipore Sigma, St. Louis, MO) as a benign
additive was applied to the surface of selected test and control coupons (according to procedures
described in Section 2.2.2.2) along with the fentanyl, and the fentanyl and benign additive were
thoroughly mixed on the coupon surface prior to the double 60-min fentanyl dwell period.
Select test coupons (wood only) included in the matrix provided in Table 3 were spiked with a
target 1 mg of fentanyl, which accounted for either a target 5% or 100% of the total challenge
applied. For samples challenged with 100% fentanyl HC1 by weight, coupons were spiked with
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only the target 1 mg of fentanyl HC1 solid. For samples challenged with 5% fentanyl HC1 by
weight, the ascorbic acid accounted for the remaining 95% of the total applied solid (i.e., 19 mg
of a total 20 mg challenge; refer to Section 2.2.2.2).
Table 3. Decontamination Efficacy Test Matrix
Test
No.
Decontaminant
Material
Surface Type(s)
Fentanyl HC1 Formulation
Dwell Time*
(min)
Replicates
Fentanyl-HCl by
Weight (%) '
Painted dry wall
Fentanyl HC1 only
60+60
3
100
1
Dahlgren Decon™
Coated steel
Fentanyl HC1 only
60+60
3
100
Wood
Fentanyl HC1 only
60+60
3
100
Wood
Fentanyl-HCl/ascorbic acid
60+60
3
5
Modified Clorox™
Painted dry wall
Fentanyl HC1 only
60+60
3
100
2
ProResults®
Coated steel
Fentanyl HC1 only
60+60
3
100
Garage and
Wood
Fentanyl HC1 only
60+60
3
100
Driveway Cleaner
Wood
Fentanyl-HCl/ascorbic acid
60+60
3
5
*: 60+60 equates to a 60-min dwell time followed by reapplication and a second 60-inin dwell time of the decontaminant.
The test matrix for short (less than 15 min) dwell times and responder gear- or PPE-related
materials is shown in Table 4. The first test was executed for three (3) materials with no
replicates to allow for multiple dwell timepoints in one test. The second and third test used a
fixed five (5)-min dwell time between the decontaminant and the fentanyl on the surface.
Table 4. Decontamination Efficacy Test Matrix for PPE Materials
Test
Decontaminant
Material
Fentanyl HC1 Formulation
Dwell Time
Replicates
No.
Surface Type(s)
(min)
Diluted Dahlgren
Decon™
Saranex®
Fentanyl HC1 only
1,2, 6, 10, 15
1
1
HazMat suit
Fentanyl HC1 only
1,2, 6, 10, 15
1
Bunker gear
Fentanyl HC1 only
1,2, 6, 10, 15
1
Neoprene
Fentanyl HC1 only
5
3
2
Diluted Dahlgren
Saranex®
Fentanyl HC1 only
5
3
Decon™
HazMat suit
Fentanyl HC1 only
5
3
Bunker gear
Fentanyl HC1 only
5
3
Modified Clorox™
Neoprene
Fentanyl HC1 only
5
3
3
ProResults®
Saranex®
Fentanyl HC1 only
5
3
Garage and
HazMat suit
Fentanyl HC1 only
5
3
Driveway Cleaner
Bunker gear
Fentanyl HC1 only
5
3
As indicated in Table 3 and Table 4, each decontaminant/material/fentanyl formulation
combination was tested in triplicate (except for the efficacy time series, Test 1 in Table 4). In
addition to the test coupons identified above, positive, blank and spike control samples were
incorporated into each test, including:
• Positive Controls - Indoor material coupons that were spiked with fentanyl (with or
without benign additive) using the same equipment and procedures as used to spike the
test coupons, but to which no decontaminant was applied. Following the fentanyl contact
and decontaminant (for test coupons) dwell periods, positive controls were extracted with
solvent, and extracts were analyzed alongside the test coupons.
• Procedural Blanks - Indoor material coupons that were not spiked with fentanyl (with or
without benign additive) but that were decontaminated, extracted with solvent, and
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analyzed alongside the test coupons using the same procedures and equipment (one
replicate per material/decontaminant combination per test).
• Laboratory Blanks - Indoor material coupons that were not spiked with fentanyl (with or
without benign additive) or decontaminated but that were extracted with solvent and
analyzed alongside the test coupons using the same procedures and equipment (one
replicate per material per test).
• Spike Control Samples - A mass of fentanyl consistent with the amount applied to the
test coupons and positive controls (target 1 mg) that was dissolved in extraction solvent
(three replicates per test; refer to Section 2.2.2.2). Spike control replicates were generated
throughout the fentanyl (with or without benign additive) spiking operation (i.e., one
spike control prior to application of fentanyl to test/positive control coupons, one in the
middle of the operation, and one after all test/positive control coupons had been spiked).
Positive controls, procedural blank samples, and laboratory blank samples consisted of coupons
of the same indoor materials of the same dimensions as the test coupons to which they were
associated.
2.2 Experimental Methods and Materials
Experimental methods and materials used to conduct the testing described in Sections 2.1.1 and
2.1.2 are described in the subsections below.
2.2.1 Coupon Materials
Method demonstration and decontamination efficacy testing were conducted using the following
types of indoor materials: painted drywall, laminate, wood, and coated steel while Saranex®
(Tychem®), a Level B HazMat suit, bunker gear, and neoprene were used as responder/PPE
materials.
Materials were cut into coupons of uniform length (4.0 cm) and width (2.5 cm). Therefore, the
top surface area to which the fentanyl (and benign additive, as required) challenge and
decontamination technologies were applied measured 10 cm2. These dimensions enabled the
coupons to fit lying flat at the bottom of the 60-mL glass jars that were used for solvent
extraction of coupons.
Coupon thicknesses were dependent upon the material type. All coupons were visually inspected
prior to use during testing to confirm the integrity and representativeness of the material.
Coupons with irregular edges and/or damaged areas were discarded. Following cutting, coupons
were cleaned using dry air to remove dust and debris prior to use in tests. Coated steel coupons
were also wiped using IPA-soaked wipes to remove any machining/cutting grease residue.
Painted drywall is a panel made of gypsum typically pressed between two thick sheets of paper
with a layer of paint coating one side of the thick sheet of paper. Painted drywall is typically
used for walls and ceilings of indoor structures. During this work, white joint tape (Sheetrock®
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brand, Lowes, Hilliard, OH) was used to simulate the thick sheet of drywall paper and was
painted with latex paint (KILZ® latex primer, Lowes, Hilliard, OH; Behr® Premium Plus
interior flat white latex paint, Home Depot, Columbus, OH) simulating the painted surface of a
drywall board. The painted joint tape was cut into individual 10 cm2 coupons.
Laminate, Formica™, Arborite™ or Garolite™ is a sheet created by combining fiber, paper
and/or fabric with epoxy or resin and set under heat. Typical uses of laminate include household
countertops and flooring. During this work, laminate coupons with a thickness of 3.2 millimeter
(mm) (0.125 inch) were used. The 24 by 24-inch Garolite™ G-10 sheets (McMaster-Carr®,
Aurora, OH) were obtained and cut into individual 10 cm2 coupons (2.5 cm by 4 cm).
Wood refers to structural wood used for framing in commercial or residential construction and is
also referred to as dimensional lumber. Douglas Fir is commonly sold and used as dimensional
lumber due to its strength, hardness, and durability. Coupons for this testing were cut from 4-
inch by 4-inch by 8-foot untreated kiln-dried Douglas Fir dimensional lumber (Home Depot,
Columbus, OH). A target coupon thickness of approximately 0.375-inches (3/8-inches) was
used. Wood surface was cross-grain and the exposed surfaces were not sanded or sealed. After
cutting to size any remaining dust was blown off with air.
Coated Steel refers to a powder-coated hard finish of steel that is similar to but generally
considered to be more durable than conventional paint. The coating is applied electrostatically as
a free-flowing dry powder and then cured under heat. The powder may be a thermoplastic or a
thermoset polymer, mainly used for coating metals, such as household appliances, aluminum
extrusions, drum hardware, and automobile, motorcycle, and bicycle parts. For this testing, black
powder-coated steel landscape edging (Lowe's, Hilliard, OH) was used as a representative
powder-coated surface. Coupons needed for this testing were cut from the edging sections.
Saranex® is the Transcendia, Inc., brand name for polyvinylidene chloride, a vinylidene chloride
homopolymer. Saranex® offers barrier protection against gases and vapors, so among many
other uses, it is often incorporated into textile laminates to produce chemical-protective clothing.
For this work, coupons were excised from Tychem® SL hooded disposable coveralls (Grainger,
Lake Forest, IL) which are constructed of Saranex® 23P film-laminated Tyvek®.
"HazMat Suit" refers to the DuraChem® 500, a National Fire Protection Agency (NFPA) 1994-
certified (2018 edition; Class 1 and Class 2) chemical, biological, radiological, nuclear (CBRN)
protective "multiuse, single-exposure" garment manufactured by Kappler, Inc. (Guntersville,
AL). Coupons cut from larger swatches of DuraChem® 500 suit material received directly from
Kappler, Inc., were included during testing.
"Bunker gear" (i.e., turnout gear) refers to the PPE worn by firefighters, most often during
structural fire operations. A typical set of turnout gear includes coat and trousers that, according
to NFPA Standard 1971, must incorporate: (1) an outer shell (typically of Nomex®/Kevlar®
construction) for heat, fire, abrasion, and chemical resistance, (2) a waterproof moisture barrier,
and (3) an inner thermal barrier. Turnout coupons used during this testing were harvested from
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the outermost layer (outer shell) of a Chieftain® 32XTM turnout coat (Grainger, Lake Forest,
IL).
Neoprene, a synthetic rubber produced by polymerization of chloroprene, is used in a wide
variety of products and applications, including gasketing and sealing of electrical enclosures,
sports, and medical equipment (e.g., joint braces and supports), wetsuits, and safety gloves.
Individual coupons for this testing were cut from a larger 12-inch by 24-inch sheet of
multipurpose neoprene rubber (0.016-inch thickness; McMaster-Carr®, Aurora, OH).
Table 5 provides a summary of test coupon information for this work.
Table 5. Coupon Materials
Material
Description
Supplier
Coupon
Thickness
(mm)
Preparation
Painted
Drywall
White joint tape, Sheetrock®
brand, item number 15335, model
number 380041;
KILZ® latex primer, item number
45548, model number 20902
Lowes
Hilliard, OH
-0.5
1. Apply one coat of latex primer;
2. Allow to dry;
3. Apply one coat of paint;
4. Allow to dry.
5. Clean using dry air to remove debris
Belir® Premium Plus Interior Flat
White Latex Paint, item number
923827
Home Depot
Columbus, OH
Laminate
Garolite™ G-10 sheet, 24" x 24",
epoxy resin with fiberglass fabric
reinforcement, item number
9910T2
McMaster-Carr
Aurora, OH
3.2
• Coupons cut from sheet
• Clean using dry air to remove debris.
Wood
4" x 4" x 8' Untreated Kiln-Dried
Douglas Fir Dimensional Lumber,
item number 137195
Home Depot
Columbus, OH
9.5
• Cut coupons to size
• Clean using dry air to remove debris
Coated
Steel
Black powder-coated steel
landscape edging section, item
number 959658
Lowes
Hilliard, OH
3.2
• Cut coupons to size
• Clean using dry air to remove debris
Saranex®
Tychem® SL coveralls (item
number 34CL41) with
elastic material (Saranex® 23P
film laminated Tyvek
construction; white)
Grainger Lake
Forest, IL
0.3
• Cut coupons to size
• Clean using dry air to remove debris
HazMat
Suit
DuraChem® 500 HazMat and
CBRN Protective Suit material
Kappler
Guntersville, AL
0.4
• Cut coupons to size
• Clean using dry air to remove debris
Bunker
gear
Chieftain® 32XTM khaki turnout
coat, item number 1370N51;
Nomex® construction and
polymer-coated Kevlar® cuff
reinforcements
Grainger
Lake Forest, IL
0.4
• Cut coupons to size
• Clean using dry air to remove debris
Neoprene
Multipurpose neoprene rubber
sheet, item number 1370N51
McMaster-Carr
0.4
• Cut coupons to size
• Clean using dry air to remove debris
2.2.2 Fentanyl
13
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2.2.2.1 Fentanyl Source
Fentanyl HC1 (3.5 grams [g]) was purchased from Cayman Chemical Company (14719, Cayman
Chemical Company, Ann Arbor, MI). All fentanyl originated from the same synthesis/production
lot. Upon receipt, fentanyl was stored at ambient laboratory temperature in accordance with
facility and DEA security and storage policies until needed for testing. Fentanyl was stored in a
single capped vial from which working quantities were drawn for use when needed.
The purity of the fentanyl received from Cayman Chemical Company was 99.59% ± 0.18%, as
provided on the certificate of analysis received with the compound. The certificate of analysis for
the fentanyl received and used for all testing that was performed is provided as Attachment A.
2.2.2.2 Fentanyl Application
Test and positive control coupons were inspected visually prior to contamination with fentanyl
and any coupons with surface anomalies were not used. Fentanyl was applied to the center of
each designated test and positive control coupon as a single (target) 1-mg pile using a 50-|iL
Drummond Series 500 Digital Microdispenser (part no. 3-000-550, Drummond, Broomall, PA)
utilizing a borosilicate capillary tube and Teflon® plunger. The fentanyl was spread over
approximately 50% of the coupon surface (as determined visually) using an antistatic spatula
(14-245-99, Fisher Scientific, Pittsburgh, PA). This spread equates to a coupon contamination
level of approximately 200 |ig/cm2 (based on the 10-cm2 coupon contamination/decontamination
surface area). Spike control samples were generated by delivering the same quantity of fentanyl
as that applied to the surface of coupons (target 1 mg) into an empty 60-mL glass extraction jar
with subsequent addition of 10 mL of IP A to dissolve the fentanyl. Following preparation, spike
controls were processed in a manner like the coupon extracts (that is, spike controls were
sonicated and aliquoted for analysis as described for coupon extracts in Section 2.2.4).
During the fentanyl/benign additive decontamination efficacy evaluation, the benign additive
ascorbic acid was applied to the surface of the 10-cm2 coupons along with fentanyl. A target 19
mg of the ascorbic acid was applied to the center of each designated test or positive control
coupon using a 100-|iL Drummond Series 500 Digital Microdispenser (3-000-575, Drummond,
Broomall, PA). A setting of 19 |iL on the 100-|iL Drummond was used to deposit (target) 19-mg
piles of ascorbic acid. A single (target) 19-mg pile was thus applied onto the surface of the
coupon in the center. Following application of the (target) 19 mg of the ascorbic acid, the (target)
1 mg of fentanyl was applied using a 50-|iL Drummond Series 500 microdispenser (as discussed
earlier). The fentanyl/ ascorbic acid applied to each coupon was then spread over approximately
50% of the coupon surface using an antistatic spatula as described above, and the two
compounds (fentanyl and ascorbic acid) were mixed concurrently with the spreading step.
14
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2.2.2.3 Fentanyl Contact Period
Following application of fentanyl, the contaminated coupons were allowed to remain undisturbed
for a 60-min fentanyl contact period. During this contact period, the coupons were subjected to
the ambient atmosphere within the test chamber. Coupons remained uncovered during the 60-
min fentanyl contact period. While temperature and RH inside the test chamber were not
controlled to a specific target, extreme conditions were avoided. Generally, test chamber
temperature ranged from 18°C to 28°C, and RH from 30% to 70%. Temperature and RH for
each test were monitored and recorded via a HOBO UX100-003 Temperature/RH datalogger
(part no. UX100-003, Onset®, Bourne, MA). Environmental data from each test are provided in
Attachment B.
2.2.3 Application of Decontamination Technologies
2.2.3.1 Meth Remover®
Meth Remover® is a formulated aqueous alkaline decontamination solution from Apple
Environmental. It is a hydrogen peroxide-based decontaminant that is intended to be
"environmentally-friendly", non-corrosive, and used for cleanup and remediation of
methamphetamine contamination. Meth Remover® is a two-component system that includes
disodium carbonate, ethanol, and a water-based buffer (Part 1), and stabilized hydrogen peroxide
(< 8%; Part 2). The decontaminant is prepared by mixing Parts 1 and 2 in equal amounts. Meth
Remover® was prepared in accordance with manufacturer instruction prior to each test during
this work.
Prior to use during decontamination efficacy tests, hydrogen peroxide concentration and pH of
the prepared Meth Remover® decontaminant were measured. Hydrogen peroxide concentration
was measured using a Hach® hydrogen peroxide test kit (HYP-1, Hach Company, Loveland,
CO), and pH was measured using a pH meter (Orion Star™ A221 pH portable meter,
STARA2210, Thermo Fisher Scientific, Waltham, MA).
2.2.3.2 Zep® Professional Stain Remover w ith Peroxide
Zep® Professional Stain Remover with Peroxide (ZEP®) is a hydrogen peroxide-based (>5% to
< 10%) cleaner intended for use on natural and synthetic textiles including carpet and upholstery.
In addition to the hydrogen peroxide active ingredient, the cleaner includes water, sodium
acrylate copolymer (film-forming agent), and ethoxylated alcohols.
Prior to use during decontamination efficacy tests, hydrogen peroxide concentration and pH of
the cleaner were measured. Hydrogen peroxide concentration was measured using a Hach®
hydrogen peroxide test kit (HYP-1, Hach Company) and pH was measured using a pH meter
(Orion Star™ A221 pH portable meter, STARA2210, Thermo Fisher Scientific, Waltham, MA).
15
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2.2.3.3 Modified Clorox™ ProResults® Garage and Driveway Cleaner
Clorox™ ProResults® Garage and Driveway Cleaner (564084310, Walmart) is a hypochlorite-
based cleaner that includes a surfactant (myristamine oxide, CAS 3332-27-2). The cleaner was
modified in pH for use as a test decontaminant. Prior to use during testing, the necessary ratio of
cleaner to vinegar (Heinz Distilled White Vinegar, 5% acidity; 700667856063, Amazon) to
water (Crystal Springs Water) required to adjust both the pH of the cleaner to 5 and the
hypochlorite concentration to 0.5% were determined. Such adjustments were intended to produce
a decontaminant like the pH 5 bleach tested previously [1], but that also included a surfactant to
promote spreading of the decontaminant across a material surface. The pH was adjusted using
vinegar (as measured using a pH meter) and then diluted as necessary using water to target a
hypochlorite concentration of 0.5% (measured using a Hach® hypochlorite test kit). A ratio of 1
part Clorox™ ProResults® Garage and Driveway Cleaner to 0.66-parts vinegar to 1.5 parts
water was determined to produce an adjusted cleaner at the pH and hypochlorite concentration
targets.
Prior to initial application of decontaminant and prior to reapplication during indoor-related
material decontamination efficacy tests (at 60 min into the total 120-min decontaminant dwell
period), hypochlorite concentration and pH of the prepared pH-adjusted surfactant bleach were
measured. Hypochlorite concentration was measured using a Hach® hypochlorite test kit (CN-
HRDT, 2687100, Hach Company) and pH was measured using a pH meter (Orion Star™ A221
pH portable meter, STARA2210, Thermo Fisher Scientific).
2.2.3.4 Dahlgren Decon™
Dahlgren Decon™ (DD-006-RTU, First Line Technology, Chantilly, VA) is a three-component
decontaminant system including water and a surfactant package (Part A), sodium hydroxide (Part
Bl), and peracetyl borate (active ingredient; Part B2; releases peracetic acid upon dissolution in
water). Normally, Part A comes as a solid and must be dissolved in water before mixing with
Parts Bl and B2, but for this testing a "ready-to-use" (RTU) version was used that provides Part
A already dissolved in water from the manufacturer.
Approximately 1 liter of Dahlgren Decon™ was prepared for use prior to each test by mixing the
three parts in accordance with directions provided by the manufacturer. Per manufacturer
direction, the prepared decontaminant must be used (i.e., applied via spray to designated
coupons, as it pertains to this testing) within 6 h of preparation. Preparation and use of Dahlgren
Decon™ adhered to this requirement during this testing. Prior to initial application of
decontaminant and prior to reapplication during indoor-related material decontamination efficacy
tests (at 60 min into the total 120-min decontaminant dwell period), peracetic acid concentration
and pH of the prepared decontaminant were measured. Peracetic acid concentration was
measured using a LaMotte test kit (7191-02, LaMotte Company, Chestertown, MD), and the pH
was measured using a pH meter (Orion Star™ A221 pH portable meter, STARA2210, Thermo
Fisher Scientific).
16
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During PPE-related material decontamination efficacy testing, Dahlgren Decon™ was diluted by
a factor of 5 in distilled water (Crystal Springs Water) prior to use. Full-strength Dahlgren
Decon™ was first prepared as described above, then 1 part (undiluted) Dahlgren Decon™ was
mixed with 4 parts water. Peracetic acid concentration and pH of the 5-fold diluted Dahlgren
Decon™ were measured as described above for full strength Dahlgren Decon™.
2.2.3.5 Decontamincmt Application
Liquid decontaminants were applied to test and control
sample coupons via moderately low flow spray using a
handheld 1-gallon pump-pressurization-style sprayer
(12U469, Grainger, Lake Forest, IL; Figure 2) equipped
with a polyethylene nozzle (2ZV94, Grainger).
The sprayer was integrated into the test chamber (nozzle
inside; tank maintained outside) to allow for moderately
low flow spray application of the decontaminants onto
coupons while still enabling operators to work with solid
fentanyl within the safety of the test chamber. The handheld
pump sprayer was selected as it is readily commercially
available, and the pump pressurization mechanism allows
for better control of spray impact pressure.
Integration of the sprayer into the test chamber involved
replacement of the sprayer extension wand with flexible
tubing that was run into the test chamber through a port on
Figure 2. Handheld pump the side walL Specifically, the spray shut-off assembly and
pressurization-style sprayer. complete nozzle assembly were removed from the extension
wand and attached to each end of a length of chemical-
resistant Versilon™ polyvinyl chloride (PVC) tubing lined with fluorinated ethylene propylene
(FEP, 6519T14, McMaster-Carr, Aurora, OH; coiled red tubing in Figure 3 below). The nozzle
assembly was then mounted to a rail installed at the top of the test chamber that allowed the
spray delivered from the nozzle to be swept from side to side. The nozzle standoff distance (i.e.,
distance from the nozzle outlet to the top surface of the coupons placed underneath) was
approximately 10.25-inches. A variable speed motorized pulley system was used to move the
nozzle across the rail at a uniform and constant rate. The sprayer was cleaned after use and in
between changes in decontamination solution. At least one liter of distilled water was passed
through the sprayer, with any residual water discharged under air pressure.
17
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Figure 3. Sprayer and test chamber setup.
During tests with the 10-cm2 coupons, test and procedural blank coupons were placed into
separate acrylic boxes (1.75-inch square by 1-inch height; part no. 3790-CL, G&G Distributors,
Saddle Brook, NJ) on top of small PP mesh disks (1.375-inch diameter, 0.05-inch thickness, cut
from larger sheet of PP mesh (part no. 9265T47, McMaster-Carr). The acrylic boxes holding
individual coupons were placed onto a tray that was positioned underneath the sprayer nozzle.
The plastic boxes containing the coupons were arranged in two rows of eight boxes as shown in
Figure 4. The sprayer nozzle stand-off distance was set such that the spray fan/cone delivered
from the nozzle extended past the outer edges of the plastic boxes placed on the tray below
within the characterized area of the spray (Figure 5).
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Figure 4. Decontaminant spray tray setup.
Fentanyl-spiked coupon
PTFE mesh
When applying decontaminant to the coupons, the sprayer was pressurized to 20 pounds per
square inch (psi), the variable speed motor was set to the necessary rate (dependent upon the
decontaminant), and the sprayer shut-off assembly (outside the chamber/hood) was actuated to
begin spray delivery. The motorized pulley system was then activated, and the nozzle (inside the
chamber) was swept from one side to the other at a rate/speed required to deliver the target
volume of decontaminant per unit area (60 |iL/cm2) to each coupon as illustrated in Figure 5.
Following application of the decontaminant, the spray was stopped, and the nozzle was returned
to the starting position on the rail. Any decontaminant dripping from the nozzle after spray had
been stopped was collected/captured so excess decontaminant did not fall on top of the coupons
below.
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Nozzle standoff distance
(approximately 10.25-inches)
Sweep speed
Figure 5. Spray application.
As discussed in Section 2.1.1.3, the sprayer pressure and sweep rate/speed necessary to deliver
60 |iL/cm2 of decontaminant to the top surface of coupons was determined prior to testing.
Determination of the necessary sprayer pressure and sweep speed was made with the 1.75 square
inch (in2) acrylic boxes present in the tray so that uniformity of the spray delivery could be
assessed by measuring the weight of liquid added to each acrylic box. Each plastic box had an
internal area (bottom internal surface) of 18.87 cm2. The target decontaminant volume delivery
of 60 |iL/cm2 would thus equate to approximately 1.13 mL of decontaminant delivered into each
box. Various combinations of sweep speeds and sprayer pressures were evaluated, and the liquid
delivered into each box was weighed to determine successful delivery of the target 1.13 mL
volume of decontaminant.
Following application, the decontaminants were allowed to remain undisturbed on the coupons
(to react with the fentanyl challenge, in the case of test coupons) for a predetermined dwell
period. Visual observation of the wetness of each coupon was recorded. The 10-cm2 coupons in
acrylic boxes were left uncovered during the decontamination dwell period. Following the
decontaminant dwell period, 10-cm2 coupons were extracted in solvent according to procedures
described in Section 2.2.4.
2.2.3.6 Decontaminant Runoff
Decontaminant that ran off the test and procedural blank coupons following spray delivery was
collected for analysis for fentanyl by GC/MS or LC-MS/MS.
During decontamination efficacy tests described in Section 2.1.2, each coupon was placed into a
separate acrylic box during application of decontaminant via spray (see Figure 5), so that the
decontaminant runoff from each coupon was segregated for collection. A PP mesh placed
underneath the coupons in the acrylic boxes provided stable elevation of the coupons off the
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bottom of the acrylic boxes to prevent the coupons from contacting any decontaminant runoff
(that may potentially contain fentanyl physically removed from the coupon by the spray-
application of decontaminant). Following removal of coupons from the acrylic boxes for
extraction with solvent (Section 2.2.4), the acrylic boxes (containing decontaminant runoff and
PP mesh) were placed into individual 250-mL glass jars (05-719-61, Fisher Scientific), and the
acrylic boxes and runoff contents were extracted with 20 mL of IP A and 5 mL 3M STS.
Following extraction of the acrylic box and runoff contents, aliquots of the IP A layers of the
extracts were transferred into individual gas chromatograph (GC) vials (21140 (vial), 24670
(cap), Fisher Scientific (Restek Corp.), and extracts were analyzed via GC/MS (Section 2.3.1) or
LC-MS/MS (Section 2.3.2).
2.2.4 Extraction of Fentanyl from Coupons
All coupons were extracted by placing each into a separate 60-mL glass jar (05-719-257, Fisher
Scientific) containing 10 mL of IP A and 5 mL of 3M STS quench. IP A was selected based on
the results of previous solvent extraction method testing [1], Using the dimensions provided in
Section 2.2.1 and Table 5, coupons of the indoor materials fit lying flat within the inside
diameter of the extraction jar identified above. The 10 mL of IP A reached a height within the jar
of approximately 1 cm. This jar and volume of solvent were sufficient to submerge all coupon
types fully. Wood coupons were placed face down as they floated to the top surface of the
solvent due to buoyancy.
Following the addition of coupons to the extraction solvent within each jar, the jars were swirled
by hand for approximately 5-10 seconds and placed into a sonicator (Branson Model 5510R-
DTH). Extraction jars were sonicated at 40 to 60 kilohertz for 10 min. Within 30 min of
completing this process, aliquots of approximately 0.5 mL from each extraction jar were
transferred to individual GC vials and sealed (21140 (GC vial), 24670 (GC vial cap), Fisher
Scientific (Restek Corp.)). Samples that were not analyzed the same day were stored at -20 ±
10°C.
2.3 Analytical Methods
As described in Section 2.1.1.4, the strategy for quantification of residual fentanyl in coupon and
decontaminant runoff extracts included both GC/MS and LC-MS/MS analyses. GC/MS was used
for quantitation of fentanyl in control samples of known concentration as well as initial analyses
of samples of unknown concentration (i.e., decontamination test samples). Samples below the
quantitation range of GC/MS were then analyzed via LC-MS/MS.
The GC/MS and LC-MS/MS analyses did not include a qualitative assessment of fentanyl
degradation byproducts and neither were other analytical methods considered. The previous
fentanyl decontamination effort [1] included some qualitative interpretation of byproduct
formation.
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2.3.1 Quantitative Fentanyl Analysis - GC/MS
Samples were analyzed in selected ion monitoring (SIM) mode on an Agilent 6890 using an
Agilent 5973A mass selective detector (MSD; Agilent Technologies, Santa Clara, CA). Modern
instrumental SIM analysis allows for multiple ion selections while still providing increased
sensitivity. Fentanyl was detected using ions m/z 245 (quantifier ion), 146, 189, 105, and 202. A
decafluorotriphenylphosphine (DFTPP) tune check was performed on the MSD to ensure proper
operation prior to sample analysis. Prior to GC/MS analysis, samples were spiked with a known
amount of fentanyl-d5 (F-001-1ML, Sigma-Aldrich) to use as an IS (m/z 250 as quantifier ion).
The concentration of analyte in samples was interpolated using the analyte area/IS area ratio and
the regression equation generated from calibration standards. See Section 4.2.3 for GC/MS
calibration details.
Table 6 provides the GC/MS conditions that were used during fentanyl analyses. Refer to
Section 4.2.3 for QA/QC provisions that were included during analyses to ensure adequate
performance of the GC/MS across the calibration range.
Table 6. GC/MS Conditions for Quantitative Fentanyl Analysis
Parameter
Description
Instrument
Agilent Model 6890 Gas Chromatograph equipped with HP 5973A
Mass Selective Detector and Model 7683 Automatic Sampler
Data System
MSD ChemStation
Column
Rxi-5Sil MS, 30.0 meters x 0.25 mm, 0.25 ^m film thickness
Liner Type
4 mm sp 1 it/sp 1 itle s s
Carrier Gas Flow rate
1.2 mL/min
Column Temperature
50 °C initial temperature, hold 0.5 min, 30 °C/min to 280 °C, hold
1.0 min
Injection Volume
3.0 jiL
Injection Temperature
250 °C
MS Quad Temperature
Ux
o
o
O
MS Source Temperature
230 °C
Solvent Delay
3.1 min
2.3.2 Quantitative Fentanyl Analysis — LC-MS/MS
Coupon extracts and aliquots of decontaminant runoff were analyzed using LC-MS/MS to
quantify the amount of residual fentanyl present. An AB Sciex 5500 triple quadrupole MS
(SCIEX, Framingham, MA) coupled to a Shimadzu 20 XR series LC (Shimadzu, Columbia,
MD) was used for sample analyses. Fentanyl was quantitated in sample extracts using a
reversed-phase high performance liquid chromatography (HPLC) method and multiple reaction
monitoring (MRM). MRM provides high specificity and sensitivity and is typically used in
quantitative applications. The MRM transition with the best signal-to-noise ratio is usually
selected for quantitation. Fentanyl-ds (F-001-1ML, Sigma-Aldrich) was used as the IS for
quantitation of fentanyl and was added to calibration standards, controls, and test samples just
prior to LC-MS/MS analysis (nominal concentration in samples after addition of 0.45 ng/mL).
Table 7 provides the ion transitions that were used for detection and quantitation of fentanyl.
22
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Table 7. LC-MS/MS Analyte Ion Transitions
Analyte
Precursor Ion
(m/z)
Product Ion Quantifier
(m/z)
Fentanyl
337
188
Fcntanyl-ds
342
188
The lower limit of quantitation for fentanyl free base was 0.010 ng/mL, which was equal to the
concentration of the lowest standard used to generate the calibration curve.
The concentration of analyte in samples was interpolated using the analyte area/IS area ratio and
the regression equation generated from calibration standards. Samples that quantitated below the
lowest calibration standard concentration or displayed area counts below the lowest
concentration on the calibration curve were reported as less than the Lower Limit of Quantitation
(LLOQ; e.g., <0.01 ng/mL). The less-than-the-LLOQ value was corrected to account for the
sample dilution factor. Samples that quantitated above the highest calibration standard were re-
diluted and reanalyzed. See Section 4.2.2 for LC-MS/MS calibration details. All data were
reported to two significant figures.
LC-MS/MS parameters that were used are provided in Table 8.
Table 8. LC-MS/MS Conditions for Quantitative Fentanyl Analysis
Parameter
Description
Ionization Mode and Polarity
Electrospray Ionization, Positive Mode
HPLC Column
Restek Allure PFPP A, 2.1 x 50 mm, 5 (un (part no. 9169552)
Column Temperature
35 °C
Curtain Gas
Nitrogen (20 psi pressure)
IonSpray Voltage
2500 V
Ion Source Temperature
500 °C
Entrance Potential
10 V
Cell Exit Potential
15 V
Mobile Phase
A: 2 mM Formic Acid/2 mM Ammonium Formate in Water
B: 2 mM Formic Acid/2 mM Ammonium Formate in Methanol
Time (min)
%B
Flow Rate
(mL/min)
0.0
20
0.5
Mobile Phase Gradient
1.0
20
0.5
2.0
100
0.7
4.0
100
0.7
4.1
20
0.5
5.0
20
0.5
Injection Volume
4nL
Run Time
5 min
A Pentafluorophenylpropyl phase.
Samples in IPA were diluted at least 10-fold prior to LC-MS/MS analysis. Samples (e.g.,
aliquots of decontaminant runoff) were matrix-matched to the calibration standards by addition
of IPA to a final concentration of approximately 10%. Alternative dilution factors were used for
23
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samples of high analyte concentration or to reduce sample matrix concentration (such as residual
Dahlgren Decon™). Sample dilutions were performed using calibrated positive displacement
pipettes and were documented on the sample chain of custody (CoC; refer to Section 4.3) and
laboratory record book (LRB).
2.4 Calculations
For each fentanyl formulation/indoor material/decontaminant combination, means of the coupon
mass recoveries, coupon residual contamination, and decontamination efficacy values were
calculated and reported, along with percent RSD.
2.4.1 Decontamination Efficacy Evaluation
Test, control, and blank coupon and runoff extract concentrations were provided in units of (_ig of
fentanyl per mL of extract by the GC/MS ChemStation software (ver. E.02.02 SP1) or in units of
ng of fentanyl HC1 per mL of extract by the LC-MS/MS Analyst® software (ver. 1.6.2) through
comparison of the analyte and IS peak areas to the GC/MS or LC-MS/MS calibration curves.
GC/MS calibration data were fitted to a quadratic regression while the LC-MS/MS calibration
data fit a linear regression (1/x2 weighting). Based on the regression, concentrations of fentanyl
in the coupon and decontaminant runoff extracts were determined (calculated by the software)
according to either Equation 1 (quadratic regression) or Equation 2 (linear regression):
Quadratic regression:
(1)
where: Aa = Analyte peak area
Ais = Internal standard peak area
Ca = Actual analyte concentration (|ig/mL)
Cis = Internal standard concentration (|ig/mL)
a, b, c = quadratic regression coefficients.
Linear regression:
(cA/cIS) ^
(2)
A IS DF
where: Aa = Analyte peak area
Ais = Internal standard peak area
b = y-intercept of regression curve
Ca = Actual analyte concentration (ng/mL)
Cis = Internal standard concentration (ng/mL)
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DF = Dilution factor (set to 1 in the software; actual dilution factor is applied in the
raw analytical data spreadsheet)
m = slope of regression curve.
In Equations 1 and 2, Ca (actual fentanyl concentration in |ig/mL (GC/MS) or ng/mL (LCMS/
MS)) is determined as fentanyl free base equivalents (since the calibrations standards are
prepared from free base fentanyl). Equation 3 was applied to the results to convert to the
equivalent fentanyl HC1 concentration:
ConcExt = CAx (^i) (3)
where: ConcExt = Coupon/runoff extract concentration in terms of fentanyl HC1 (ng/mL
(LC-MS/MS) or |ig/mL (GC/MS))
Ca = Coupon/runoff extract concentration (fentanyl free base equivalents) provided
by the LC-MS/MS software (ng/mL) or GC/MS software (|ig/mL)
MWhci = Fentanyl HC1 molecular weight (372.94 g/mol)
MWFree = Free base fentanyl molecular weight (336.47 g/mol).
Mass recovered from the coupons or runoff samples via extraction was determined according to
Equation 4:
MassRec = ConCE^Xnl°lE" (4)
where: MassRec = Fentanyl mass recovered from the coupon/runoff (|ig)
ConcExt = Coupon/runoff extract concentration in terms of fentanyl HC1 (ng/mL
(LC-MS/MS) or |ig/mL (GC/MS))
Vol Ext = Volume of coupon/runoff extraction solvent (mL)
Conv = Conversion factor (1000 for LC-MS/MS analyses; 1 for GC/MS analyses)
Residual fentanyl contamination for each coupon was determined using the calculated mass
recovered from the coupon and the coupon contamination/decontamination surface area,
according to Equation 5:
4- Massac
ContRes = (5)
ACoupon
where: ContRes = Residual coupon contamination (|ig/cm2)
MassRec = Fentanyl mass recovered (|ig)
Acoupon = Contamination/decontamination surface area of the coupon (cm2).
Total sample mass was determined using the masses recovered from extraction of the coupon
and extraction of the associated runoff sample, according to Equation 6:
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MdSSj0i — MaSSRec (coupon) McLSSrbc (runoff)
where: Massiot = Total fentanyl mass recovered (|ig)
MassRec (coupon) = Fentanyl mass recovered from the coupon (|ig)
MassRec (runoff) = Fentanyl mass recovered from the runoff (|ig)
Percent efficacy of decontamination from each individual test coupon or percent total efficacy
for each coupon/runoff combination was calculated according to Equation 7:
Efficacy = (x 100o/o (7)
V MassRec (pos) )
where: Mass(x) = Either MassRec (coupon) or Massiot from Equation 4 (|ig)
MassRec (pos) = Fentanyl mass recovered from the associated positive control (|ig).
Calculation of efficacy using MassRec (coupon) provided a measurement of the ability of the
decontaminant to remove fentanyl contamination from the surface of the material coupons, either
by chemical decontamination of fentanyl or by physical removal. Calculation of efficacy using
Massiot intended to decouple physical removal from the efficacy calculation and provide an
indication of the ability of the decontaminant to chemically degrade fentanyl contamination.
2.4.2 Decontamination Efficacy Evaluation (Benign Additive Ascorbic Acid)
Fentanyl mass recoveries, residual contamination, total sample masses, and percent efficacies
were calculated according to Equations 1 through 7 used during the initial fentanyl
decontamination efficacy evaluations (10-cm2 coupons, fentanyl HC1 only without benign
additives present; Section 2.1.2).
2.5 Statistical Analyses
For each test condition as defined by the decontamination technology/material
type/decontamination period/challenge additive combinations, mean and percent RSD of the
fentanyl recovery from test coupon and positive control sample sets were calculated, and test
coupon fentanyl recovery means were compared to associated positive control means to
determine if statistically significant decontamination of fentanyl occurred. Geometric means
were compared for trial datasets as shown in Tables 3 and 4; and arithmetic means were
compared for datasets as shown in Table 2 to be consistent with the transformations used in the
comparisons between test conditions described below. These decontamination comparisons were
conducted both between mean positive control coupon mass versus mean test coupon extracted
mass, and between mean positive control coupon mass and the sum of test coupon extracted
mass and the decontaminant runoff mass.
F-tests were used to determine if the variances of the set of three test coupon mass (|ig) results
were equal to the set of three positive control coupon mass (|ig) results. The null hypothesis that
the variances of the two sets were equal was rejected if the F-test p-value was < 0.05. One-tailed,
26
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two-sample Student's Mests (homoscedastic or heteroscedastic based on the F-test result) were
then used to determine if the means of the test results were significantly less than the positive
controls or not [3], The null hypothesis that the test coupon and positive control coupon means
were equal was rejected if the Mest p-value was < 0.05. If multiple pairwise comparisons are
performed at a 0.05 significance level, the probability of falsely rejecting a true null hypothesis
at least once over all tests is greater than 0.05. Bonferroni corrections for multiple comparisons
were therefore applied within each trial dataset to maintain a familywise error rate of 0.05 over
all tests within a given dataset and test coupon outcome measurement [4], Rejecting the null
hypothesis represents evidence that fentanyl mass was reduced after the application of the
decontaminant.
Additionally, four separate groups of analyses were conducted to test whether there were
significant differences in fentanyl recovery between the test conditions of interest. Tukey's
multiple comparisons procedure was performed following each of the analyses where more than
two conditions were compared [5], Like the Bonferroni procedure, Tukey's procedure adjusts the
p-values of the pairwise comparisons to maintain a familywise error rate of 0.05 per each one-
way analysis of variance (ANOVA) model over the multiple comparisons being performed.
Tukey's procedure was selected to account for multiple comparisons instead of the Bonferroni
corrections because Tukey's procedure typically has a higher power to detect differences
between conditions but applies only when all pairwise comparisons are made within a model.
The Tukey-adjusted p-values are presented only if significant differences were identified.
Within each condition, the characteristics of fentanyl application to the positive controls are
assumed to be the same as the characteristics of application to the test coupons with regard to
variability from coupon to coupon and in the average amount of fentanyl applied. Acceptance
criteria for the spike control results (average within 80% to 120% of theoretical, <30% RSD) are
intended to support this assumption. For accurate comparison of the performance of the
decontaminants within each material as described above, the amount of fentanyl applied to the
test coupons must be consistent across all conditions being compared within a given analysis. To
evaluate consistency of fentanyl application across the three samples per material of each
decontaminant, the 58 comparisons described above for the test coupons were repeated using the
total mass recoveries from the positive control sets of three samples associated with each of the
31 test conditions.
2.5.1 Group 1: Comparisons of Decontaminant Performance within Material Type
For the first group of ANOVA model with an effect for material (painted dry wall paper, powder-
coated steel, and wood for 60 + 60-min decontamination periods; bunker gear, HazMat suit,
neoprene and Saranex® for 5-min decontamination periods) was fitted separately to each of four
decontaminant and decontamination period combinations (Dahlgren Decon™ at 60 + 60 min,
Diluted Dahlgren Decon™ at 5 min, pH 5 modified surfactant bleach at 60 + 60 min, pH 5
modified surfactant bleach at 5 min) to determine if there were significant performance
differences among the different materials. Materials were challenged with a targeted 1 mg of
27
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fentanyl. Tukey's multiple comparisons procedure was performed for the three (for the 60 + 60-
min decontamination periods) or six (for 5-min decontamination periods) possible pairwise
comparisons between the three or four materials within each decontaminant/decontamination
period group to determine which pairs of materials had mean total mass recoveries that were
significantly different from each other.
2.5.2 Group 2: Decontamination Period Effect Analysis Plan
For the second group of analyses, a one-way ANOVA model with an effect for decontamination
period (60 min, 60 + 60 min) was fitted separately to each of two decontaminants (Dahlgren
Decon™, pH 5 modified surfactant bleach) to determine if there were significant performance
differences depending on decontamination time. All data for the 60-min decontamination period
were taken from the previous study [1], while all data for the 60 + 60-min decontamination
period were taken from the current study. Materials were challenged with a targeted 1 mg of
fentanyl. The effect of decontamination period was calculated while collapsing across material
condition based on the assumption that there is no material effect. The results of Analysis 1 and
from the previous study suggested that there was not a statistically significant effect of material
on fentanyl recovery within the group of materials tested for each study. However, it should be
noted the materials used in the current study (with a decontamination period of 60 + 60 min)
were different from the materials used in the previous study (with a decontamination period of
60 min). It was therefore impossible to isolate the material effect from the decontamination
period effect for these materials, and impossible to confirm the assumption of no material effect
when comparing acrylic, laminate, painted drywall, and stainless steel (previous study materials
[1]) to drywall paper, powder-coated steel, and wood (current study materials).
2.5.3 Group 3: Challenge Additive Effect Analysis Plan
For the third group of analyses, a one-way ANOVA model with an effect for challenge
compound (1 mg fentanyl, 1 mg fentanyl +19 mg ascorbic acid) was fitted separately to each of
two decontaminants (Dahlgren Decon™, pH 5 modified surfactant bleach) to determine if there
were significant performance differences when the challenge did contain ascorbic acid versus did
not contain ascorbic acid. Only the wood material was challenged with both fentanyl and
fentanyl + ascorbic acid. Therefore, the model was only fitted to data from the wood material.
The decontamination period for all conditions was 60 min.
2.5.4 Group 4: Decontaminant Effect Analysis Plan
For the fourth group of analyses, a one-way ANOVA model with an effect for decontaminant
across the two fentanyl decontamination studies (Dahlgren Decon™, EasyDecon DF200,
OxiClean™, pH 7 bleach, pH 5 bleach, pH 5 modified surfactant bleach, and water from the
previous study [1]; Meth Remover® and ZEP® from the current study) was fitted separately for
each positive control and test coupon sample set from the laminate material condition to
determine if there were significant performance differences among the decontaminants tested.
28
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Only the laminate material was selected because this was the only material overlapping between
the previous and current decontamination study. Materials were challenged with a targeted 1 mg
of fentanyl and underwent a 60-min decontamination period.
For all four analysis groups (Sections 2.5.1 - 2.5.4), positive control coupon fentanyl mass (|ig)
and test coupon total sample fentanyl mass (|ig) were evaluated to determine if the total mass
recovery data were reasonably normally distributed or if a natural logarithmic transformation
would improve adherence to the statistical assumptions of normality and equal variances. The
total sample fentanyl mass was calculated as the sum of the coupon extract mass and the
decontaminant runoff mass. For the positive control data, a logarithmic transformation improved
conformity to the assumptions of normality and equal variance in all but one case. Therefore,
fentanyl recovery masses for all positive control samples were log-transformed. For the test
sample data, a natural logarithmic transformation also improved conformity to the assumption of
normality and equal variances for almost all datasets in analysis groups 1-3. Therefore, the test
sample data were log-transformed for the analyses in groups 1-3. However, the data better
conformed to the assumptions of normality and equal variances when untransformed for the
fourth analysis group, which examined the effect of decontaminant for the laminate material. For
this reason and to maintain consistency with the analysis in the previous study, data were left
untransformed for the fourth analysis group.
29
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RESULTS
3.1 Methods Demonstration
3.1.1 Fentanyl Delivery (Spiking) Characterization
Under the previous fentanyl decontamination effort [1], the 50-|iL Drummond Series 500 Digital
Microdispenser with a setting of 1.9 |iL on the 50-|iL Drummond produced generally accurate
and repeatable target 1 mg masses of fentanyl, producing an average percent recovery of 95%
with ±14% RSD. This setting was utilized without further verification prior to the method
development and decontamination testing. During the method development and decontamination
testing, spike controls exceeded the 80% to 120% of the target application. The Drummond
setting was gradually adjusted downwards between decontamination tests to get the delivered
fentanyl mass within the desired range.
3.1.2 Decontaminant Spray Delivery Characterization
As discussed in Section 2.1.1.3, the sprayer system used to apply the test decontaminants was
characterized using each specific decontamination technology to determine system settings
necessary to deliver the decontaminants at the target application volume of 60 |iL/cm2.
Empty acrylic runoff boxes (refer to Section 2.2.3.5) were arranged underneath the sprayer as
depicted in Figure 6. As described in Section 2.2.3.5, the acrylic boxes were weighed before and
after spray delivery of the decontaminants to determine the mass, and thus volume per unit area,
of decontaminant delivered.
¦4 Sprayer travel direction
1
2
3
4
5
6
7
S
9
10
11
12
13
14
15
16
Figure 6. Test sample arrangement under sprayer.
Spray system settings necessary for delivery of each decontaminant at the target volume of 60
|iL/cm2 are summarized in Table 9.
30
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Table 9. Decontaminant Spray Application Settings
Decontaminant
No. of
Sprayer
Passes
Sprayer
Motor
Setting
Pass
Speed
(cm/s)
Sprayer
Pressure
(psi)
Plastic
Box
Area
(cm2)
Target
Decontaminant
Delivery
(jiL/cm2)
Target
Decontaminant
Weight
(g)
Meth Remover®
76
7.9
1.16
ZEP®
70
6.9
1.16
pH 5 Modified
Surfactant Bleach
1
56
4.6
20
18.87
60
1.13
Dahlgren Decon™
84
9.1
1.28
Diluted Dahlgren
Decon™
75
7.9
1.16
Replicate decontaminant weights measured during characterization, average weights,
variabilities (% relative standard deviation [RSD]), and delivery accuracy (percent of target) for
each decontaminant are provided in Tables CI through C5 in Attachment C. A summary of the
delivery accuracies for each decontaminant is provided in Table 10.
Table 10. Decontaminant Spray Delivery Summary
Decontaminant
Average Percent
Lowest Percent of
Highest Percent of
of Target
Target (Position)
Target (Position)
(% ± SD)
(%)
(%)
Meth Remover®
111 ± 4
104 (#11)
117 (#13)
ZEP®
95 ± 10
83 (#11)
112 (#6)
pH 5 Modified Surfactant Bleach
99 ±3
93 (#9)
104 (#14)
Dahlgren Decon™
103 ±6
92 (#3)
115(#10)
Diluted Dahlgren Decon™
98 ±6
86 (#12)
107 (#1)
Results showed that the delivery of the decontaminant was accurate and precise with no evidence
for a nonuniform spatial distribution across the 16 coupon locations.
3.1.3 Quench Method Demonstration
The quench method test matrix was intended to demonstrate the adequacy of 3M STS as a
quench agent for halting decontamination of fentanyl by the hydrogen peroxide-based
decontaminants (Meth Remover® and ZEP®).
As described in Section 2.1.1.4, representative decontaminant runoff sample extracts were
prepared by adding 0.99 mL of test decontaminant to 20 mL of IP A with 5 mL of 3M STS
solution. Samples were produced in triplicate, and the extracts were post-spiked with a dilute
solution of fentanyl. Recovery of fentanyl > 70% of the theoretical post-spiked amount in
representative test extract matrix samples (quench samples) during the 72-h delayed analyses of
the samples and conformance to the QA/QC criteria for fentanyl-ds IS response discussed in
Section 4.2.2 and provided in Table 36 would demonstrate the adequacy of the quench method
and that no interferences due to the sample matrices were occurring. Results of the quench
method scoping test are summarized in Table 11 and Figures 7 and 8.
31
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Table 11. Quench Method Demonstration Test, Average Mass Recovery
Quench Method Demonstration
40 ng target fentanyl mass, GC/MS
analysis, 5 mL 3M STS quench
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The results suggest that addition of 5 mL of 3M STS with the extraction solvent may be an
effective quench for both hydrogen peroxide-containing decontaminants. Recoveries of post-
spiked fentanyl in samples containing Meth Remover® and ZEP® ranged from 91% to 92% for
the GC/MS results and ranged from 100% to 105% for the LC-MS/MS results, respectively,
which satisfy the minimum criterion of 70%. Thus, the results suggested that the defined quench
and sample storage procedures (addition of 5 mL of 3M STS to the IPA used to extract coupon
and runoff samples and storage of samples at -20°C for up to three days prior to GC/MS or LC-
MS/MS analysis) were adequate for preservation of the mass of fentanyl that was post-spiked
into the representative coupon and runoff samples extracts.
3.2 Decontamination Efficacy Evaluation - Building Materials
3.2.1 Hydrogen Peroxide-Based Decontaminants
The tests described in Table 2 in Section 2.1.2 evaluated the efficacy of two (2) decontaminants
(Meth Remover® and ZEP®) to decontaminate fentanyl on the surface of 10-cm2 coupons of
four (4) materials (painted drywall paper, powder-coated steel, laminate, and wood). Each test
included the necessary replicate test and control samples to evaluate efficacy of a single
decontaminant on all four material types.
Average spike control recoveries for each test are provided in Table 12. The average amounts
recovered exceeded the 80% to 120% of the target application, which may be due to a tighter
packing of the fentanyl in the vial leading to higher amounts spiked at the same setting on the
Drummond Pipettor from the previous research effort.
Table 12. Decontamination Efficacy Testing, Spike Control Average Recovery
Test Number
Test
Avg Mass
(fig)
% RSD
Avg % Recovery
(vs theoretical)
1
Meth Remover®
1234
63
122%
2
ZEP®
1398
206
138%
No fentanyl was detected in any laboratory blank samples. Fentanyl was detected in all
procedural blank coupon extracts and in runoff samples for all decontaminants, but all were
below the criteria provided in Table 34 in Section 4.1. Detections in procedural blank coupon
extract and runoff samples were always less than 1% of the associated positive controls for all
materials and both decontaminants.
Mass recovery results are summarized in Figure 9. Average fentanyl mass recoveries, standard
deviations, and variabilities (%RSD) for replicate test and positive control coupons of all four
material types for each decontaminant are provided in Tables D1 and D2 of Attachment D.
Average total test sample mass recoveries (mass recovered from extraction of the coupon sample
plus mass recovered from extraction of the associated runoff) and percent of the total test sample
mass recovery versus the associated positive controls are provided as well. In some instances,
solid material (which could potentially be undissolved fentanyl) was observed on the surface of
33
-------
replicate coupons following the 60-min decontaminant dwell period and prior to solvent
extraction of the coupons.
Average Fentanyl Mass Recovery
10-cm2 coupons, fentanyl HCI, 60 minute decontamination dwell time
(error bars equal ± one standard deviation)
MR: Meth Remover® ; ZEP: Zep® Professional Stain Remover with Peroxide
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Average Percent Efficacy
10-cm2 coupons, fentanyl HCI, 60 minute decontamination dwell time
MR: Meth Remover® ; ZEP: Zep® Professional Stain Remover with Peroxide
100%
¦ Physical and chemical decontamination
LU
PM
Decontaminant, Material
Figure 10. Decontamination efficacy testing, average percent efficacy.
3.2.2 Decontamination Efficacy Evaluation — Reapplication of Decontaminants
The two tests described in Table 3 in Section 2.1.2 evaluated the efficacy of two (2)
decontaminants (Dahlgren Decon™ and pH 5 modified surfactant bleach) to decontaminate
fentanyl directly on the surface of 10-cm2 coupons. For these two tests, the recovered fentanyl
amounts are for the test coupons to which the decontaminant was applied twice, each with a 60-
min dwell time. Each test included three (3) materials (painted drywall paper, powder-coated
steel, and wood) with a fourth coupon consisting of the same wood but in the presence of
ascorbic acid as a benign additive to the fentanyl on the surface. Each test included the necessary
replicate test and control samples to evaluate efficacy of a single decontaminant on all material
types.
Average spike control recoveries for each test are provided in Table 13. The average amounts
recovered exceeded the 80% to 120% of the target application, which may be due to a tighter
packing of the fentanyl in the vial leading to higher amounts spiked at the same setting on the
Drummond Pipettor from the previous research effort.
35
-------
Table 13. Decontamination Efficacy Testing, Spike Control Average Recovery
Test Number
Test
Spike
Type
Avg Mass
(Mg)
%
RSD
Avg % Recovery
(vs theoretical)
1
Dahlgren Decon™
No AA
1549
10%
153%
With AA
1703
5.6%
168%
2
pH 5 modified
No AA
1509
10%
149%
surfactant bleach
With AA
1377
6.6%
136%
AA: Ascorbic Acid
No fentanyl was detected in any laboratory blank samples. Fentanyl was also not detected in the
procedural blank coupon extracts and in runoff samples associated with the Dahlgren Decon™
applications. For the pH 5 modified surfactant bleach decontamination test, fentanyl was
detected in the procedural blanks, but all were below the criteria provided in Table 34 in Section
4.1. Detections in procedural blank coupon extracts and runoff samples were always less than
1% of the associated positive controls for all materials and pH 5 modified surfactant bleach.
Mass recovery results are summarized in Figure 11. Average fentanyl mass recoveries, standard
deviations, and variabilities (%RSD) for replicate test and positive control coupons of all three
material types and the fourth material with ascorbic acid for each decontaminant are provided in
Tables D4 and D5, Attachment D. Average total test sample mass recoveries (mass recovered
from extraction of the coupon sample plus mass recovered from extraction of the associated
runoff) and percent of the total test sample mass recovery versus the associated positive controls
are provided as well. In some instances, solid material (which could potentially be undissolved
fentanyl) was observed on the surface of replicate coupons following the first 60-min
decontaminant dwell time or after the double 60-min decontaminant dwell periods and prior to
solvent extraction of the coupons.
36
-------
Average Fentanyl Mass Recovery
10-cm2 coupons, fentanyl HCI, 60 + 60 minute decontamination dwell time
(error bars equal ± one standard deviation)
AA: Ascorbic Acid
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Figure 11. Decontamination efficacy testing, average mass recovery.
Efficacy for each of the decontaminants was calculated by comparing the residual fentanyl mass
on the test coupons against the fentanyl mass recovered from the associated positive control.
Once again, two efficacy values were calculated - one using only the masses recovered from
extraction of the test and positive control coupons (efficacy thus does not differentiate between
physical removal and chemical decontamination), and another value wherein the fentanyl mass
measured in the runoff extract was added to the test sample coupon mass before comparison to
the positive control (to attempt to decouple physical removal from chemical decontamination).
Average percent efficacies (both excluding and including average runoff mass) for each
material/decontaminant combination are summarized in Figure 12. Efficacy values are also
tabulated in Table D6, Attachment D.
Efficacies ranged from 88% to more than 99.9% and from 80% more than 99.9% for the
Dahlgren Decon™ and modified pH 5 modified surfactant bleach products, respectively, when
considering only the chemical degradation by the decontaminant. Recovered fentanyl mass was
higher in the presence of ascorbic acid.
37
-------
Average Percent Efficacy
10-cm2 coupons, fentanyl HCI, 60 + 60 minute decontamination dwell time
AA: Ascorbic Acid
¦ Physical and chemical decontamination
¦ Chemical decontamination only
Decontaminant, Material
Figure 12. Decontamination efficacy testing, average percent efficacy.
3.2.3 Decontamination Efficacy Evaluation — PPE/Gear Materials
The first test described in Table 4 in Section 2.1.2 evaluated the efficacy of dilute Dahlgren
Decon™ as a function of time (1-15 min) for three materials (Saranex®, HazMat suit, and
bunker gear). Average mass recovered for the spike controls (n=3) was 1366 |ig with a 14%
RSD. As mentioned before, the amount spiked was higher (135% of theoretical mass spiked)
than expected.
No fentanyl was detected in any laboratory or procedural blank samples taken at the longest 15-
min dwell time.
Recovered fentanyl mass (no replicates) for the five timepoints (coupon, runoff, and sum) are
tabulated in Table 14. Positive controls are reported as recovered mass at 0 min (start fentanyl
mass on material).
38
-------
Table 14. Average Mass Recovery, Diluted Dahlgren Decon™
Time
Saranex®
Ha/Mat suit
Bunker gear
(min)
Coupon
Mass
(fig)
Runoff
Mass
(fig)
Sum
(fig)
Coupon
Mass
(fig)
Runoff
Mass
(fig)
Sum
(fig)
Coupon
Mass
(fig)
Runoff
Mass
(fig)
Sum
(fig)
0
1205
-
1205
1360
-
1360
1127
-
1127
1
36
4.1
40
11
2.2
13
15
4.6
19
3
0.63
2.0
2.7
14
0.67
15
316 A
2.0
318
6
0.58
7.2
7.8
47 A
1.4
48
25
5.5
31
10
4.6
2.8
7.5
11
1.5
12
41 A
2.3
43
15
20
1.9
22
0.31
0.53
0.84
61 A
0.87
62
A Solid material observed on coupon surface following application.
Recovered fentanyl mass from coupon and runoff combined was in general less than 50 |ig
except for one outlier of 318 |ig which was accompanied by observed solid material remaining
on the coupon surface. A recovered total mass of 50 |ig equates to approximately a 96% efficacy,
which can be reached within minutes following interaction with the diluted Dahlgren Decon™
decontaminant on these gear/PPE materials.
The second and third tests described in Table 4 in Section 2.1.2 evaluated the efficacy of two (2)
decontaminants (Dahlgren Decon™ and pH 5 modified surfactant bleach) with a 5-min dwell
time of the solution to decontaminate fentanyl directly on the surface of 10-cm2 gear/PPE
material coupons. Each test included the necessary replicate test and control samples to evaluate
efficacy of a single decontaminant on all material types.
Average spike control recoveries for each test are provided in Table 15. The average amounts
recovered slightly exceeded the 80% to 120% of the target application, which may be due to a
tighter packing of the fentanyl in the vial leading to higher amounts spiked. However, this
exceedance of the target application does not impact the decontamination process itself as the
amount of decontaminant significantly exceeds the amount of fentanyl on the material.
Table 15. Decontamination Efficacy Testing, Spike Control Average Recovery
Test
Number
Test
Avg Mass
(Mg)
% RSD
Avg % Recovery
(vs theoretical)
1
pH 5 modified surfactant bleach
1280
15%
130%
2
Diluted Dahlgren Decon™
1261
25%
128%
No fentanyl was detected in any laboratory blank samples. Fentanyl was also not detected in the
procedural blank coupon extracts and in runoff samples associated with the Dahlgren Decon™
applications. For the pH 5 modified surfactant bleach decontamination test, fentanyl was
detected in the procedural blanks, but all were well below the criteria provided in Table 34 in
Section 4.1. Detections in procedural blank coupon extract and runoff samples were always less
than 0.1% of the associated positive controls for all materials and pH 5 modified surfactant
bleach.
Mass recovery results are summarized in Figure 13. Average fentanyl mass recoveries, standard
deviations, and variabilities (%RSD) for replicate test and positive control coupons of all four
39
-------
PPE/gear material types are provided in Tables D7 and D8 in Attachment D. Average total test
sample mass recoveries (mass recovered from extraction of the coupon sample plus mass
recovered from extraction of the associated runoff) and percent of the total test sample mass
recovery versus the associated positive controls are provided as well. As expected, based on
previous research results, in some instances, solid material (which could potentially be
undissolved fentanyl) was observed on the surface of replicate coupons following the 5-min
decontaminant dwell time and prior to solvent extraction of the coupons
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Average Percent Efficacy
10-cm2 coupons, fentanyl HCI, 5 minute decontamination dwell time
pH adjusted bleach with surfactant Diluted Dahlgren Decon
¦ Physical and chemical decontamination
¦ Chemical decontamination only
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surfactant bleach with Surfactant (they do not share a similarity designation). However, the
positive control means were not significantly different for Water and pH 5 modified surfactant
bleach (they share the "B" similarity designation).
3.3.1.1 Group 1 Material Effect Analysis Effects
There were no significant differences in fentanyl recovery mass for the positive controls between
any pairs of materials within a given decontaminant trial (Dahlgren Decon™ at 60 + 60 min,
Diluted Dahlgren Decon™ at 5 min, or pH 5 bleach with surfactant at 60 + 60 min and 5 min).
Refer to Tables 16 through 19.
3.3.1.2 Group 2 Decontamination Period Effect Analysis Results
Collapsing across materials, positive control fentanyl recovery masses for a 60-min
decontamination period [1] were found to be statistically significantly different from the fentanyl
recovery masses of a 60 + 60-min decontamination period for both the Dahlgren Decon™ and
the pH 5 modified surfactant bleach decontaminants. We can therefore not say that fentanyl was
applied equally across the two fentanyl decontamination studies materials. Refer to Tables 20
and 21.
3.3.1.3 Group 3 Challenge Additive Effect Analysis Results
Positive control materials challenged with 1 mg fentanyl only were found to have a significantly
different fentanyl recovery mass than materials challenged with 1 mg fentanyl +19 mg ascorbic
acid for the Dahlgren Decon™ test condition. We cannot say that fentanyl was applied equally in
the 1-mg fentanyl and the 1-mg fentanyl + 19 mg ascorbic acid test conditions with Dahlgren
Decon™. Positive control fentanyl recovery mass was not found to be different for the 1 mg
fentanyl challenge compound versus the 1 mg fentanyl +19 mg ascorbic acid compound for the
pH 5 modified surfactant bleach decontaminant test condition. Refer to Tables 22 and 23.
3.3.1.4 Group 4 Decontaminant Effect Analysis Results
In the previous fentanyl decontamination study [1], only EasyDecon DF200 and pH 5 modified
surfactant bleach resulted in significantly different fentanyl recovery masses for the positive
control samples on the laminate material. The positive control geometric mean recovery masses
for EasyDecon DF200 and pH 5 modified surfactant bleach, which were identified as
significantly different in the previous study, remained significantly different for the current
analysis. Additionally, the positive control fentanyl recovery mass for Meth Remover® differs
significantly from the positive control fentanyl recovery mass of EasyDecon DF200, and
fentanyl recovery mass for ZEP® differs significantly from the positive control fentanyl
recovery mass of seven of the nine other decontaminant positive controls. We cannot conclude
that fentanyl was applied equally across the laminate in both studies. Refer to Table 24.
42
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Group 1:
Table 16. ANOVA Results for Dahlgren Decon™ at 60 + 60 min (Positive Controls)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery (jug)
Tukey-Adjusted
p-Value *
Dahlgren Decon™
Wood
60+60
fentanyl
A
1267
No significant
differences.
Dahlgren Decon™
Painted drywall
60+60
fentanyl
A
1368
Dahlgren Decon™
Coated steel
60+60
fentanyl
A
1668
* There were no significant differences between any pairs of materials.
Table 17. ANOVA Results for Diluted Dahlgren Decon™ at 5 min (Positive Controls)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery (jug)
Tukey-Adjusted
p-Value *
Diluted Dahlgren Decon™
Neoprene
5
fentanyl
A
818
Diluted Dahlgren Decon™
Bunker gear
5
fentanyl
A
916
No significant
Diluted Dahlgren Decon™
HazMat suit
5
fentanyl
A
991
differences.
Diluted Dahlgren Decon™
Saranex®
5
fentanyl
A
1150
* There were no significant differences between any pairs of materials.
Table 18. ANOVA Results for pH 5 Modified Surfactant Bleach at 60 + 60 nun (Positive Controls)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(WS)
Tukey-Adjusted
p-Value *
pH 5 modified surfactant bleach
Wood
60 + 60
fentanyl
A
1203
No significant
differences.
pH 5 modified surfactant bleach
Painted drywall
60 + 60
fentanyl
A
1208
pH 5 modified surfactant bleach
Coated steel
60 + 60
fentanyl
A
1208
* There were no significant differences between any pairs of materials.
Table 19. ANOVA Results for pH 5 Modified Surfactant Bleach at 5 ntin (Positive Controls)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(MS)
Tukey-Adjusted
p-Value *
pH 5 modified surfactant bleach
Saranex®
5
fentanyl
A
1020
No significant
differences.
pH 5 modified surfactant bleach
HazMat suit
5
fentanyl
A
1068
pH 5 modified surfactant bleach
Bunker gear
5
fentanyl
A
1129
pH 5 modified surfactant bleach
Neoprene
5
fentanyl
A
1138
* There were no significant differences between any pairs of materials.
43
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Group 2:
Table 20. ANOVA Results for Dahlgren Decon™, Collapsed over Multiple Materials (Positive Controls)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(Mg)
p-Value
Dahlgren Decon™ *
Multiple
60
fentanyl
A
859
<0.0001 (60 min <
60 + 60 min)
Dahlgren Decon™
Multiple
60 + 60
fentanyl
B
1425
* Data from previous study [1].
Table 21. ANOVA Results for pH 5 Modified Surfactant Bleach, Collapsed over Multiple Materials (Positive Controls)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery (jug)
p-Value
pH 5 modified surfactant bleach *
Multiple
60
fentanyl
A
947
<0.0001 (60 min<
60 + 60 min)
pH 5 modified surfactant bleach
Multiple
60+60
fentanyl
B
1206
* Data from previous study [1].
Group 3:
Table 22. ANOVA Results for Dahlgren Decon™ on Wood at 60 + 60 min (Positive Controls)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(Mg)
p-Value
Dahlgren Decon™
Wood
60 + 60
fentanyl
A
1267
0.0464 (fentanyl <
fentanyl + ascorbic acid)
Dahlgren Decon™
Wood
60 + 60
fentanyl + ascorbic acid
B
1406
Table 23. ANOVA Results for pH 5 Modified Surfactant Bleach on Wood at 60 + 60 min (Positive Controls)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(Mg)
p-Value *
pH 5 modified surfactant bleach
Wood
60 + 60
fentanyl
A
1203
No significant
differences.
pH 5 modified surfactant bleach
Wood
60 + 60
fentanyl + ascorbic acid
A
1273
44
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Group 4:
Table 24. ANOVA Results on Laminate at 60 nun (Positive Controls)
Decontaminant
Material
Period
(min)
Challenge
Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(MS)
Tukey-Adjusted p-Value
EasyDecon DF200*
Laminate
60
fentanyl
A
736
0.0337 (DF200 < Meth Remover®)
OxiClean™ *
Laminate
60
fentanyl
A
796
0.0215 (DF200 < pH 5 bleach)
0.0039 (DF200 < ZEP®)
pH 7 bleach*
Laminate
60
fentanyl
A
820
Dahlgren Decon™ *
Laminate
60
fentanyl
A
824
0.0140 (OxiClean™ < ZEP®)
Water*
Laminate
60
fentanyl
AB
849
0.0247 (Dahlgren Decon™ < ZEP®)
pH 5 modified surfactant bleach
Laminate
60
fentanyl
BCD
854
0.0398 (Water < ZEP®)
Meth Remover®
Laminate
60
fentanyl
CD
1174
0.0229 (pH 7 bleach < ZEP®)
pH 5 bleach*
Laminate
60
fentanyl
DE
1208
ZEP®
Laminate
60
fentanyl
E
1341
0.0429 (pH 5 modified surfactant bleach < ZEP®)
* Data from previous study [1].
45
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3.3.2 Comparison of Test Sample Results
Tables 25 through 33 present the mean mass recoveries of the ANOVA models for each of the
analysis groups ordered from lowest to highest mean, along with the significant Tukey-adjusted
comparisons. The estimated geometric mean is presented in Tables 25 through 32, corresponding
to the natural log-transformed data in analyses for Group 1 through Group 3. The estimated
arithmetic mean is presented in Table 33, corresponding to the untransformed data for analysis of
Group 4. As in Tables 16 through 24, the characters in the "Similarity Designation" column
indicate the statistical similarity of the mean total mass recovery of a given condition to the mean
total mass recovery of the other conditions tested. All rows with the same similarity designation
value are not statistically significantly different from each other.
3.3.2.1 Group 1 Material Effect Results
There were no significant differences in fentanyl recovery mass between any pairs of materials
within a given decontaminant (Dahlgren Decon™ at 60 + 60 min, Diluted Dahlgren Decon™ at
5 min, or pH 5 modified surfactant bleach with surfactant at 60 + 60 min and 5 min). Refer to
Tables 25 through 28. Hence, there are no significant impacts of the materials on the efficacies
within the limitations of this study.
3.3.2.2 Group 2 Decontamination Period Effect Analysis Results
Collapsing across materials, fentanyl recovery mass for a 60-min decontamination period was
not found to be statistically significantly different from the fentanyl recovery mass of a 60 + 60
min decontamination period for the Dahlgren Decon™ or pH 5 modified surfactant bleach
decontaminants. The reason that no significant differences were observed may be questioned,
however, due to significant differences between positive control masses for the 60 min and 60 +
60 min test conditions (see Limitations Section 3.3.3). Refer to Tables 29 and 30.
3.3.2.3 Group 3 Challenge Additive Effect Analysis Results
Materials challenged with 1 mg fentanyl only were found to have a significantly lower fentanyl
recovery mass than materials challenged with 1 mg fentanyl +19 mg ascorbic acid for the
Dahlgren Decon™ test condition. The cause of this significant difference may be questioned,
however, due to significant differences between positive control masses for the 1 mg fentanyl
and 1 mg fentanyl + 19 mg ascorbic acid conditions (see Limitations Section 3.3.3). Fentanyl
recovery mass was not found to be different for the 1 mg fentanyl challenge compound versus
the 1 mg fentanyl +19 mg ascorbic acid compound for the pH 5 modified surfactant bleach
decontaminant. Refer to Tables 31 and 32.
3.3.2.4 Group 4 Decontaminant Effect Results
In the previous decontamination study [1], Water and OxiClean™ had the highest fentanyl
recovery masses of any of the seven decontaminants tested and were the most statistically
46
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different of the decontaminants. Water had been significantly different from four and
OxiClean™ had been significantly different from five of the other decontaminants tested on
laminate.
For the current analysis, Meth Remover® and ZEP® numerically resulted in the highest fentanyl
recovery masses. Meth Remover® was significantly different from five of the other eight
decontaminants tested (all decontaminants except for Water, OxiClean™, and ZEP®). ZEP®
was different from all other decontaminants tested except for Meth Remover®. The cause of
significant differences between Meth Remover® or ZEP® and the decontaminants from the
previous study [1] may be questioned, however, due to significant differences between the
positive control masses for Meth Remover® and ZEP® compared to the remaining
decontaminants. Refer to Table 33.
47
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Group 1:
Table 25. ANOVA Results for Dahlgren Decon™ at 60 + 60 min (Test Samples)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(MS)
Tukey-Adjusted
p-Value *
Dahlgren Decon™
Painted drywall
60+60
fentanyl
A
2
No significant
differences.
Dahlgren Decon™
Coated steel
60+60
fentanyl
A
2
Dahlgren Decon™
Wood
60+60
fentanyl
A
2
* There were no significant differences between any pairs of materials.
Table 26. ANOVA Results for Diluted Dahlgren Decon™ at 5 min (Test Samples)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
fog)
Tukey-Adjusted
p-Value *
Diluted Dahlgren Decon™
Saranex®
5
fentanyl
A
12
Diluted Dahlgren Decon™
Neoprene
5
fentanyl
A
15
No significant
Diluted Dahlgren Decon™
Bunker gear
5
fentanyl
A
32
differences.
Diluted Dahlgren Decon™
HazMat suit
5
fentanyl
A
86
* There were no significant differences between any pairs of materials.
Table 27. ANOVA Results for pH 5 Modified Surfactant Bleach at 60 + 60 min (Test Samples)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(Mg)
Tukey-Adjusted
p-Value *
pH 5 modified surfactant bleach
Painted drywall
60 + 60
fentanyl
A
40
No significant
differences.
pH 5 modified surfactant bleach
Coated steel
60 + 60
fentanyl
A
77
pH 5 modified surfactant bleach
Wood
60 + 60
fentanyl
A
118
* There were no significant differences between any pairs of materials.
48
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Table 28. ANOVA Results for pH 5 Modified Surfactant Bleach at 5 min (Test Samples)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(Mg)
Tukey-Adjusted
p-Value *
pH 5 modified surfactant bleach
Bunker gear
5
fentanyl
A
389
pH 5 modified surfactant bleach
HazMat suit
5
fentanyl
A
422
No significant
pH 5 modified surfactant bleach
Neoprene
5
fentanyl
A
437
differences.
pH 5 modified surfactant bleach
Saranex®
5
fentanyl
A
444
* There were no significant differences between any pairs of materials.
Group 2:
Table 29. ANOVA Results for Dahlgren Decon™, Collapsed over Multiple Materials (Test Samples)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(Mg)
p-Value *
Dahlgren Decon™
Multiple
60 + 60
fentanyl
A
2
No significant
Dahlgren Decon™ **
Multiple
60
fentanyl
A
5
differences.
* There were no significant differences between the decontamination periods.
** Data from previous study [1].
Table 30. ANOVA Results for pH 5 Modified Surfactant Bleach, Collapsed over Multiple Materials (Test Samples)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(Mg)
p-Value *
pH 5 modified surfactant bleach **
Multiple
60
fentanyl
A
35
No significant
differences.
pH 5 modified surfactant bleach
Multiple
60+60
fentanyl
A
71
* There were no significant differences between the decontamination periods.
** Data from previous study [1].
49
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Group 3:
Table 31. ANOVA Results for Dahlgren Decon™ on Wood at 60 + 60 min (Test Samples)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(MS)
p-Value
Dahlgren Decon™
Wood
60 + 60
fentanyl
A
2
0.0124 (fentanyl <
fentanyl + ascorbic acid)
Dahlgren Decon™
Wood
60 + 60
fentanyl + ascorbic acid
B
33
Table 32. ANOVA Results for pH 5 Modified Surfactant Bleach on Wood at 60 + 60 min (Test Samples)
Decontaminant
Material
Period
(min)
Challenge Compound
Similarity
Designation
Geometric Mean
Mass Recovery
(MS)
p-Value *
pH 5 modified surfactant bleach
Wood
60 + 60
fentanyl
A
118
No significant
differences.
pH 5 modified surfactant bleach
Wood
60 + 60
fentanyl + ascorbic acid
A
233
* There were no significant differences between the challenge compounds.
50
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Group 4:
Table 33. ANOVA Results on Laminate at 60 min (Test Samples)
Decontaminant
Material
Period
(min)
Challenge
Compound
Similarity
Designation
Arithmetic Mean
Mass Recovery
(MS)
Tukey-Adjusted p-Value
pH 5 modified surfactant bleach
Laminate
60
fentanyl
A
64
0.0050 (pH 5 modified surfactant bleach < Water)
0.0010 (pH 5 modified surfactant bleach < OxiClean™)
0.0002 (pH 5 modified surfactant bleach < Meth Remover®)
EasyDecon DF200*
Laminate
60
fentanyl
A
72
<0.0001 (pH 5 modified surfactant bleach < ZEP®)
0.0056 (DF200 < Water)
pH 5 bleach*
Laminate
60
fentanyl
A
102
0.0011 (DF200 < OxiClean™)
0.0002 (DF200 < Meth Remover®)
0.0001 (DF200 < ZEP®)
Dahlgren Decon™ *
Laminate
60
fentanyl
A
114
0.0088 (pH 5 bleach < Water)
0.0017 (pH 5 bleach < OxiClean™)
pH 7 bleach*
Laminate
60
fentanyl
AB
223
0.0003 (pH 5 bleach < Meth Remover®)
<0.0001 (pH 5 bleach < ZEP®)
Water*
Laminate
60
fentanyl
BCD
628
0.0106 (Dahlgren Decon™ < Water)
0.0020 (Dahlgren Decon™ < OxiClean™)
0.0004 (Dahlgren Decon™ < Meth Remover®)
OxiClean™ *
Laminate
60
fentanyl
CD
739
<0.0001 (Dahlgren Decon™ < ZEP®)
0.0102 (pH 7 bleach < OxiClean™)
Meth Remover®
Laminate
60
fentanyl
DE
818
0.0024 (pH 7 bleach < Meth Remover®)
<0.0001 (pH 7 bleach < ZEP®)
ZEP®
Laminate
60
fentanyl
E
1168
0.0064 (Water < ZEP®)
0.0421 (OxiClean™ < ZEP®)
* Data from previous study [1 .
51
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3.3.3 Limitations of Statistical A nalysis
The significant differences between the positive controls for the various test conditions is an
important limitation for the current analysis, especially when comparing the results obtained in
this study against the results from previous study [1], The positive control fentanyl recovery
masses differed significantly for 11 of the 58 total comparisons with 8 of the 36 comparisons
between the two studies, suggesting that fentanyl application was mostly inconsistent across the
two studies. This inconsistency could result in artificial differences being detected if conditions
that were otherwise equal received significantly different amounts of fentanyl and could also
conceal differences between conditions if conditions that truly have lower fentanyl recovery
mass received more fentanyl and conditions that truly had greater fentanyl recovery mass
received less.
Further, the assumption of no material effect that was made to justify collapsing the data across
materials in the analysis of Group 2 could not be completely assessed. While there is not strong
evidence of differences between material groups within the past study [1] and results discussed
here, the materials tested at the 60-min decontamination period [1] differed from the materials
tested at a 60 + 60-min period. Therefore, the differences between the materials in the 60-min
condition [1] and 60 + 60-min condition could not be assessed separately from the differences
between the decontamination periods. The lack of significant differences between 60 versus 60 +
60-min decontamination periods could be due to no true differences between decontamination
periods or could be due to material differences and decontamination period differences balancing
out.
52
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QUALITY ASSURANCE/QUALITY CONTROL
Quality objectives and performance criteria described in the sections below provide the
requirements for determining the adequacy of data generated during this project. Methods were
considered acceptable and valid data were assumed if the data quality objectives for the test
measurements were met, and the Technical Systems Audit (TSA), Performance Evaluation (PE),
and data quality audits demonstrated acceptable results, as described in Sections 4.5, 4.6, and
4.7. Accuracy was ensured by the calibration of the instruments. The PE audits further confirmed
that valid data were generated (refer to Section 4.6). The consistently higher amounts of spiked
fentanyl based on the recovered mass from the spike controls did not impact the results of this
study. The only difficulty was that a direct comparison of the collected data (amounts recovered
from positive controls and test coupons) against the results from the previous study was more
complicated.
4.1 Data Quality Indicators
Data quality indicators and results are provided in Table 34. In general, the data quality indicator
results were acceptable per the Quality Assurance Project Plan (QAPP) titled Quality Assurance
Project Plan for Remediation of fentanyl Contaminated Indoor Environments (version 1.0, 20
February 2020), as amended, including checks of the measurement methods for temperature, RH,
time, volume, mass, fentanyl recovery from blank samples and spike controls, and pH.
Attainment of these data quality indicator results limited the amount of error introduced into the
evaluation results except for the amount of fentanyl recovered in the spike control (SC) extracts
from multiple tests.
Table 34. Data Quality Indicators and Results
Measurement
Parameter
Method
Data Quality Indicators
Results
Temperature
(°C)
National
Institute of
Standards and
Testing (NIST)-
traceable
thermometer
Compare against calibrated
thermometer once before testing;
agree ± 1 °C through 60 min.
The HOBO UX100 datalogger used in the test chamber remained within
0.1°C of the calibrated reference through 1 h.
Relative
Humidity
(%)'
NIST-traceable
hygrometer
Compare against calibrated
hygrometer once before testing; agree
±10% through 60 min.
The HOBO UX100 datalogger used in the test chamber during the TO
remained within 1.9% of the calibrated reference through one hour.
Time
(seconds,
sec)
Timer/data
logger
Compare to time provided at
NIST.time.gov once before testing;
agree ±2 sec/h.
No difference was observed between the timer and NIST.time.gov after 1 h.
Volume
(mL, jiL)
Calibrated
pipette (LC-
MS/MS sample
dilution)
Pipettes were checked for accuracy
and repeatability once before use by
determining mass of water delivered.
The syringe/pipette was acceptable if
the range of observed masses for five
replicate droplets was ±10% of
expected.
Five pipettes used for LC-MS/MS sample dilution were checked.
Systematic and random percent error ranges for each are provided below:
• Pipette 1 at 1, 5, and 10 jxL - 0.18% to 7.7%
• Pipette 2 at 3, 10, and 25 jiL - 0.34% to 8.0%
• Pipette 3 at 20, 35, and 50 jxL - 0.00% to 1.2%
• Pipette 4 at 50, 100, and 250 }iL - 0.12% to 1.2%
• Pipette 5 at 100, 500, and 1,000 jiL - 0.16% to 1.2%
53
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Parameter
Measurement
Method
Data Quality Indicators
Results
Volume
(mL, jiL)
Pump
pressurization-
style sprayer
(decontaminant
delivery)
Sweep speed and sprayer pressure for
the sprayer nozzle to achieve the 600
p.L/coupon target application volume
was determined once prior to testing
by weighing the amount of
decontaminant delivered. Spray
procedures were acceptable if the
range of measured volumes for 5
applications is ± 20% of the nominal
target volume.
Three (3) replicate spray applications were delivered to acrylic runoff boxes
at all sixteen (16) positions across the characterized spray area. Average
spray delivery across the 16 positions across the 3 replicates were:
• Meth Remover® - 111% of theoretical
• ZEP® - 95% of theoretical
• pH 5 modified surfactant bleach - 99% of theoretical
• - 103% of theoretical
• Diluted Dahlgren Decon™ - 98% of theoretical
Detailed data are provided in Section 3.1.2.
Fentanyl in
Laboratory
Blank
Coupon
Extracts
(Hg/mL)
Extraction,
LC/MS/MS or
GC/MS
Laboratory blanks (coupons without
applied fentanyl that are not
decontaminated) should have less than
50% of the lowest detected amount on
the test coupon or 1% of the amount
on the positive controls, whichever is
lower.
No fentanyl outside the stated criteria was measured in any laboratory blank
sample extract throughout testing.
Fentanyl in
Procedural
Blank
Coupon
Extracts
(Hg/mL)
Extraction,
LC/MS/MS or
GC/MS
Procedural blanks (coupons without
applied fentanyl that are
decontaminated) should have less than
50% of the lowest detected amount on
the test coupon or 1% of the amount
on the positive controls, whichever is
lower.
No fentanyl outside the stated criteria was measured in any procedural blank
sample extracts throughout testing.
Fentanyl in
Spike
Control
Extracts
(Hg/mL)
LC/MS/MS or
GC/MS
The mean of the spike controls
included with each day of testing was
within 80% to 120% of the target
application and had a CoV of <30%
between replicates.
Mean and RSD of the following SC sets were outside tolerance:
• Test 1 of Table 12, 122% avg SC recovery, 5.1% RSD
• Test 2 of Table 12, 138% avg SC recovery, 15% RSD
• Test 1 of Table 13 (no ascorbic acid), 153% avg SC recovery, 10% RSD
• Test 1 of Table 13 (with ascorbic acid), 168% avg SC recovery, 5.6% RSD
• Test 2 of Table 13 (no ascorbic acid), 149% avg SC recovery, 10% RSD
• Test 2 of Table 13 (with ascorbic acid), 136% avg SC recovery, 6.6% RSD
• Time dependence test (Section 3.2.3), 135% avg SC recovery, 14% RSD
• Test 1 of Table 15, 130% avg SC recovery, 15% RSD
• Test 2 of Table 15, 128% avg SC recovery, 25% RSD
PH
Calibrated pH
meter
Meter was checked for accuracy prior
to each use using unexpired buffer
solutions at:
• pH 4 (SB101-500, Fisher Scientific)
• pH 7 (1552-16, Fisher Scientific)
• pH 10 (1602-16, Fisher Scientific)
• pH 12.5 (1618-16, Fisher Scientific)
Check value must be within ±0.1 pH
units of the buffer value.
Meter was checked before each use using the specified unexpired buffer
solutions and was within tolerance during all checks.
4.2 Instrument Calibration
4.2.1 Calibration Schedules
Instrumentation needed for the investigation was maintained and operated according to the
quality and safety requirements and documentation of Battelle's HMRC. Except for the GC/MS
and LC-MS/MS, all instruments utilized during the project were calibrated as stipulated by the
manufacturer or, at a minimum, annually. The GC/MS and LC-MS/MS were calibrated as
described in Sections 4.2.2 and 4.2.3. Table 35 provides calibration schedules for instruments
that were used during the evaluation.
54
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Table 35. Equipment Calibration Schedule
Equipment
Frequency
Calibrated pipettes
Prior to the investigation and annually thereafter. Calibration/accuracy was
also verified as described in Table 34.
Calibrated UX100 HOBO
Hygrometer/Thermometer
Prior to the investigation by the manufacturer. After the manufacturer-
provided calibration expired, use of the expired unit was discontinued and
the unit was discarded. A new manufacturer-calibrated unit was obtained
for use.
Timer
Prior to the investigation by the manufacturer. After the manufacturer-
provided calibration expired, use of the expired unit was discontinued and
the unit was discarded. A new manufacturer-calibrated unit was obtained
for use.
LC-MS/MS
Calibrated prior to analysis of each set of test samples (calibration curve)
and a calibration verification standard was analyzed after every ten samples
(see Section 4.2.2).
GC/MS
Calibrated prior to analysis of each set of test samples (calibration curve)
and a calibration verification standard was analyzed after every ten samples
(see Section 4.2.3).
pH meter
Prior to the investigation and annually thereafter. Calibration/accuracy was
verified prior to each use as described in Table 34.
4.2.2 LC-MS/MS Calibration
Fentanyl (certified analytical reference material; separate source from the fentanyl used to
contaminate test coupons; part numbers F-013-1ML and F-002-1ML, Sigma Aldrich) was used
to create calibration standards (concentrations corrected for percent purity; see Section 2.2.2.1)
encompassing the appropriate analysis range. Calibration standards were kept and used for no
longer than two months from the date of creation. After two months of use, calibration standards
and continuing calibration verifications (CCVs) were replaced with a new (fresh) set prepared from
an unopened stock ampoule. The old and new sets were then analyzed, and the results were
compared to ensure consistency, accuracy, and precision (in terms of the criteria provided in
Table 36) and to demonstrate that degradation of the old standards during the two-month period
of use had not occurred. In all cases, calibration standard and CCV concentrations remained
stable (i.e., no degradation occurred) during the two-month use period.
A seven-point calibration curve for fentanyl was used with a lower calibration level of 0.010
ng/mL and an upper limit of 5.0 ng/mL. A linear or quadratic regression (specified in the raw
data product) was used to describe the data with 1/x2 weighting. The origin was not included for
regression. Limits were also placed on the percent bias (Equation 8) observed in the standards.
Bias = x 100% (8)
where: Ev = expected value from calibration curve
Ov = observed value from standard
55
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The percent bias for the low standard had to be less than or equal to 25%, and the percent bias
for the remaining standards had to be less than or equal to 15%. The signal-to-noise ratio of the
lowest calibration standard had to be approximately 3:1 at minimum. The retention time (RT) for
each target compound and IS in each injection had to be within ±0.1 min of the RT for the same
components in the mid-level calibration standard.
Solvent blank and double blank samples were included during analytical runs to confirm that no
fentanyl carryover occurred. Solvent blank sample analysis results had to be below the value of
the lowest calibration standard.
Independently prepared CCVs were analyzed prior to sample analysis, following every ten (or
fewer) test/control samples (not including blanks or matrix samples), and at the end of each set
of samples. Two CCV concentrations were used, one of which was equal to the low calibration
standard (0.010 ng/mL) and the other within the calibration range (2 ng/mL). CCV response had
to be within 25% of the nominal concentration for the low level CCV used and within 15% of the
nominal concentration for the mid-range CCV for fentanyl analyses to be considered acceptable.
Calibration standards and CCVs were matched to the samples undergoing analysis as closely as
possible. For example, test samples in IPA prepared for analysis by a 10-fold dilution in water
were quantitated using standards and CCVs prepared in 10% IPA.
The area of fentanyl-ds IS in the test samples was compared to the area of fentanyl-ds IS in the
nearest passing calibration standard or passing CCV. Fentanyl-ds area in the test samples had to
fall within 50% to 200% of the area of the IS in the calibration standard or CCV to which it was
compared (criteria per EPA Method 8000D [6]). As described in Section 2.1.1.4
(Decontamination Technology Quench and Matrix Effect Evaluation), the validity of the
assumption that any test sample matrix would affect analysis of fentanyl and fentanyl-ds IS in an
identical manner was evaluated prior to decontamination efficacy testing. Based on the data and
criteria (refer to Section 3.1.3) the assumption held, so IS response variability within the range of
50% to 200% of that of the nearest passing calibration standard or CCV was considered
acceptable and IS was assumed to properly compensate for identical effects on fentanyl response
due to sample matrices. In certain cases, IS area was found to be outside this acceptance range,
so the test sample dilution factor was increased to reduce the effect of sample matrix.
Table 36 summarizes LC-MS/MS analysis performance parameters and acceptance criteria.
56
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Table 36. LC-MS/MS Analysis Performance Parameters and Acceptance Criteria
Parameter
Criterion
Coefficient of determination (r2)
>0.990
% Bias for the lowest calibration standard
< 25%
% Bias for remaining calibration standards (except lowest standard)
< 15%
Solvent blank samples
< lowest calibration standard
% Bias for the low CCV
< 25%
% Bias for the high CCV
< 15%
Signal-to-noise ratio for the lowest calibration standard
Minimum of 3:1
RT for target compound and IS
±0.1 min. as same compounds in
mid-level calibration standard
Fentanyl-ds IS area in samples
50% to 200% area of nearest passing
calibration standard or passing CCV
4.2.3 GC/MS Calibration
As with LC-MS/MS calibration, fentanyl (certified analytical reference material; separate source
from the fentanyl used to contaminate test and control coupons; part numbers F-013-1ML and F-
002-1ML, Sigma Aldrich) were used to create calibration standards encompassing the
appropriate analysis range. Use and retention schedules and replacement procedures for
calibration standards and CCVs for GC/MS calibration were identical to those described for LC-
MS/MS calibration standards and CCVs in Section 4.2.2. Calibration standards and CCVs were
stored in a freezer at -20 ± 10°C when not in use. A five-point calibration for fentanyl was used
with a lower calibration level of 0.25 |ig/mL and an upper limit of 25 [j,g/mL. As discussed in
Section 2.3.1, fentanyl-d5 was used as an IS during GC/MS fentanyl analyses, and the IS was
added to samples just prior to GC/MS analyses. Target fentanyl-d5 concentration in samples was
5 [^g/mL. Fentanyl-d5 IS area in the test samples was compared to the area of fentanyl-d5 IS in
the nearest passing calibration standard or passing CC V and IS acceptance criteria was identical
to that described for acceptance of fentanyl-d5 IS response during LC-MS/MS analyses in
Section 4.2.2 and indicated in Table 36.
A quadratic regression curve fit was applied to the calibration data. As during LC-MS/MS
calibration (as indicated in Table 36), the GC/MS was recalibrated if the r2 from the regression
analysis of the standards was less than 0.990. Limits were also placed on the percent bias
(Equation 8) observed in the standards. As required during LC-MS/MS analyses (as described in
Section 4.4.2 and indicated in Table 36), the percent bias for the low standard must be less than
or equal to 25%, and the percent bias for the remaining standards must be less than or equal to
15%. The GC/MS was tuned initially and as needed following manufacturer's guidelines. A tune
check was performed before running each set of samples using DFTPP. A 12-h tune time was
not employed.
Following analysis of the calibration standards at the beginning of each analytical run, a solvent
blank sample was analyzed to confirm that no fentanyl carryover was occurring. Solvent blank
sample analysis results were always below the value of the lowest calibration standard. As with
LCMS/MS analysis, independently prepared CCV standards were analyzed prior to sample
57
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analysis, following every five test/control samples and at the end of each set of samples. Use of
CCVs and CCV acceptance criteria during GC/MS analyses were identical to those described for
LC-MS/MS analyses in Section 4.4.2 and summarized in Table 36.
4.3 Sample Handling and Custody
At all times during the project, protocols required by the U.S. DEA and Battelle's HMRC were
followed in the movement and use of fentanyl within the test facility. CoC forms were used to
ensure that test samples generated during the work were traceable throughout all phases of
testing.
4.4 Test Parameter Control Sheets
Test measurements and information were recorded on test parameter control sheets (TPCSs) or
in an LRB. Monitoring of test conditions, parameters, and times was performed by technical staff
familiar with the QAPP and testing and was documented on the TPCS.
4.5 Technical Systems Audit
The QA Officer performed a TSA at the HMRC facility in West Jefferson, Ohio, during
decontamination efficacy testing on July 13, 2020. The purpose of the TSA was to ensure that
testing was performed in accordance with the QAPP. The Battelle QA Officer reviewed the
investigation methods, compared test procedures to those specified in the QAPP, and reviewed
data acquisition and handling procedures. The Battelle QA Officer did not identify any findings
that required corrective action.
4.6 Performance Evaluation Audits
PE audits, provided in Table 37 with results, addressed those reference measurements that
factored into the data used in quantitative analysis during the evaluation, including volume,
mass, and time measurements and GC/MS or LC-MS/MS calibration and performance. The mass
of fentanyl dispensed correlated directly to the mass of fentanyl on the coupons. The measured
times that fentanyl and the decontamination technologies were allowed to remain in contact with
the coupons directly influenced efficacy of the decontaminants. Calibration of the GC/MS and
LC-MS/MS and IS recovery provided confidence that the analysis system was providing
accurate data.
Temperature and RH were measured and recorded on each day of testing, but not monitored or
controlled. Therefore, no PE audit of these parameters was performed. See Attachment B for a
summary table of measured temperature and RH ranges.
During the decontaminant spray delivery characterization (Section 3.1.2), two minor spills and
one loss of a sample were reported (Tables C1-C3, Attachment C) leading to a lower average and
larger standard deviation in two of the average weights while for the third lost sample, the
average was taken over two instead of three measurements. The spray delivery characterization
58
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experiment was not repeated as the impact of these three spills was negligible. Despite the lower
measured average weight, the percent of target weight was more than 100%.
Table 3 7. Performance Evaluation Audit Results
Parameter
Audit Procedure
Required Tolerance
Results
Volume
(mL, jaL)
Pipettes were checked for accuracy and
repeatability one time before use by
determining the mass of water delivered.
The pipette was acceptable if the range
of observed masses for five droplets is
±10% of expected.
±10%
Five pipettes used for LC-MS/MS sample dilution were checked.
Systematic and random percent error ranges for each are provided below:
• Pipette 1 at 1, 5, and 10 jxL - 0.18% to 7.7%
• Pipette 2 at 3, 10, and 25 jxL - 0.34% to 8.0%
• Pipette 3 at 20, 35, and 50 }iL - 0.00% to 1.2%
• Pipette 4 at 50, 100, and 250 jxL - 0.12% to 1.2%
• Pipette 5 at 100, 500, and 1,000 jiL- 0.16% to 1.2%
Time (sec)
Compare to time provided at
NIST.time.gov once before testing; agree
±2 seconds/h.
±2 seconds/h
No difference was observed between the timer and NIST.time.gov after 1 h.
Fentanyl in
Spike
Control
Extracts
(Hg/mL)
Use LC-MS/MS to determine mass of
fentanyl delivered as a 1 mg pile into 10
mL of extraction solvent and compare to
target application level.
>80% of spike target
< 120% of spike target
< 30% CoV
Mean and RSD of the following SC sets were outside of tolerance:
• Test 1 of Table 12, 122% avg SC recovery, 5.1% RSD
• Test 2 of Table 12, 138% avg SC recovery, 15% RSD
• Test 1 of Table 13 (no ascorbic acid), 153% avg SC recovery, 10% RSD
• Test 1 of Table 13 (with ascorbic acid), 168% avg SC recovery, 5.6% RSD
• Test 2 of Table 13 (no ascorbic acid), 149% avg SC recovery, 10% RSD
• Test 2 of Table 13 (with ascorbic acid), 136% avg SC recovery, 6.6% RSD
• Time dependence test (Section 3.2.3), 135% avg SC recovery, 14% RSD
• Test 1 of Table 15, 130% avg SC recovery, 15% RSD
• Test 2 of Table 15, 128% avg SC recovery, 25% RSD
LC-MS/MS
Fentanyl
Calibration
Standards
(%)
Verify all standards and CCVs used to
calibrate and confirm calibration of the
LC-MS/MS system used for analysis fall
within the requirements provided in
Section 4.2.2.
Refer to Table 36
All standards and CCVs were within specification for all reported data.
Fentanyl-ds
IS Recovery
Use LC-MS/MS to measure from a
secondary source and compare to the
primary source one time.
±10%
IS used during analyses was compared to a secondary source. Five (5)
replicate analyses of a 1 ng/mL standard prepared from each source were
conducted. 2.2% relative percent difference in mean areas obtained.
PH
Meter was checked for accuracy prior to
each use using unexpired buffer
solutions at:
• pH 4 (SB101-500, Fisher Scientific)
• pH 7 (1552-16, Fisher Scientific),
• pH 10 (1602-16, Fisher Scientific),
• pH 12.5 (1618-16, Fisher Scientific)
±0.1 pH
Meter was checked before each use using the specified unexpired buffer
solutions and was within tolerance during all checks.
4.7 Data Quality Audit
Validation of the data included verification of the completeness of the data, compliance with the
acceptance criteria in the QAPP, recalculation checks, and tracing of the data from instrument
outputs through the final report. One hundred percent (100%) of the data was reviewed prior to
use in calculations or any data manipulation, and review was completed before the data were
provided to QA for the data quality audit.
The QA Manager, operating independently of the laboratory testing effort, audited at least 10%
of the data generated during testing. Data were traced from initial acquisition through reduction
and to final reporting. All calculations were checked.
Through the data quality audit, the TSA, and review of reports, the QA Manager ensured that
data generated during testing were valid, meeting the requirements of the QAPP.
59
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4.8 QAPP Amendments
Two (2) amendments to the QAPP were prepared during the project:
• Amendment 1 (dated June 18, 2020) revised the test matrix provided as Table 2 for the
indoor-related material decontamination efficacy evaluation described in Section B. 1.2 of
the QAPP.
• Amendment 2 (dated September 17, 2020) revised the test matrix provided as Table 3 for
the first responder PPE material-related decontamination efficacy evaluation described in
Section B.1.3 of the QAPP. An additional (third) PPE material-related decontamination
efficacy test was added to the matrix.
4.9 QAPP Deviations
Two (2) deviations from the procedures defined in the QAPP were noted during the TO:
• Per Sections B.2.3.5 and B.2.3.6 of the QAPP, the 10-cm2 test and control coupons are
placed into separate acrylic boxes during each test to collect any decontaminant (and
fentanyl) that runs off the coupon following spray-application of decontaminant. Plastic
mesh disks were placed underneath the coupons in the acrylic boxes to elevate the
coupons and prevent contact of the coupons with the decontaminant runoff that is
collected. The QAPP indicates that the plastic mesh disks will be made of
polytetrafluoroethylene (PTFE), but mesh disks made of polypropylene (PP) were used
during all tests. The PP mesh disks were cut from a larger sheet of PP mesh (part number
9265T47, McMaster-Carr) using a 1.5-inch diameter die.
• Table 8 in Section B .5 of the QAPP indicates that five (5) replicate decontaminant spray
applications will be performed during determination of necessary spray parameters for
each decontaminant to achieve the target application volume of 600 [xL/coupon (60
[xL/cm2). During characterization of the sprayer using the test decontaminants, only three
(3) replicate decontaminant spray applications were performed for each decontaminant.
60
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DISCUSSION/CONCLUSIONS
Bench scale decontamination efficacy tests were performed in which candidate decontaminants
were assessed for efficacy in decontamination of fentanyl on the surface of 10-cm2 material
coupons. Decontaminants included Meth Remover® (hydrogen peroxide active ingredient),
Zep® Professional Stain Remover with Peroxide (hydrogen peroxide), Dahlgren Decon™
(activated peracetic acid), and pH 5 modified surfactant bleach (nominal hypochlorite
concentration of 5%) derived from Clorox™ ProResults® Garage and Driveway Cleaner.
Decontaminants were applied via spray at a target application volume of 60 |iL/cm2.
Decontaminant that ran off the coupon surface during and after spray-application was collected
to assess the runoff of fentanyl from the material coupons in addition to chemical
decontamination of fentanyl.
Decontamination efficacies were calculated by both including and excluding mass detected in the
decontaminant runoff. Efficacy calculated without consideration of mass present in the runoff
describes the performance of the decontaminant with regard to both propensity for physical
removal of fentanyl by the decontaminant during spray-application as well as by chemical
degradation of fentanyl. Conversely, addition of fentanyl runoff mass to the mass recovered from
coupons via solvent extraction enables calculation of efficacy that is attributable primarily to
degradation of fentanyl.
5.1 Building Material Decontamination
After a 1-h dwell time with the fentanyl-contaminated surfaces, the Meth Remover® and ZEP®
product demonstrated similar average efficacies of 62% and 65%, respectively, across the four
materials attributable to a combination of physical removal and chemical decontamination. The
average percent efficacy dropped to 42% and 26%, respectively, across the four materials when
physical removal was decoupled from the efficacy calculation. While the application of these
two, hydrogen peroxide-containing, products led to degradation of fentanyl, their efficacies are
not as high as seen for some of the other decontamination solutions. It is possible that a longer
contact time or repeated application would improve efficacy. However, such a study was not
conducted. The Meth Remover® product was included in the completed remediation of a
fentanyl-contaminated home [2], While this suggests that residual fentanyl would have been
present after the application (and 1-h dwell time) of this product, it should be noted that the
remediation effort of the home included multiple applications of wiping and water rinsing of the
surfaces in addition to the Meth Remover® product application. The combination of the physical
removal and chemical degradation appears to have led to the successful remediation. Any future
cleanups should consider the combination of physical removal and chemical degradation.
After a double application and a total 2-h dwell time, the pH 5 modified surfactant bleach
decontaminant demonstrated average efficacies of 95% across the four materials attributable to a
combination of physical removal and chemical decontamination. The average percent efficacy
dropped to 91% across the three materials when physical removal was decoupled from the
61
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efficacy calculation. The addition of ascorbic acid mixed with the fentanyl lowered the efficacy
on wood from 84% to 80% based on chemical degradation only.
Under the same test conditions, the Dahlgren Decon™ decontaminant demonstrated an average
efficacy of 99.91% across the four materials attributable to a combination of physical removal
and chemical decontamination and the average percent efficacy dropped to 99.86% across the
four materials when physical removal was decoupled from the efficacy calculation. The addition
of ascorbic acid mixed with the fentanyl lowered the efficacy on wood from 99.8% to 97% based
on chemical degradation only.
A direct comparison between the double application data and the single application data [1] is
difficult to make as the amount of fentanyl that was applied was significantly different with more
fentanyl applied in the current study. Since efficacy values did not improve significantly, it
appears that a reapplication of these two products may not be useful unless there is evidence that
a double application can overcome any material demand of the decontamination solution. The
presence of additives such as ascorbic acid may result in lower efficacy due to a higher demand
of the decontaminant.
5.2 PPE/Responder Gear Decontamination
The Dahlgren Decon™ and pH 5 modified surfactant bleach decontaminants were also
considered in efficacy studies with a short 5-min dwell time, simulating a short dwell time as
part of decontamination line procedures.
After a 5-min dwell time, the pH 5 modified surfactant bleach decontaminant demonstrated
efficacies of 86% across the four PPE/responder gear materials attributable to a combination of
physical removal and chemical decontamination. The average percent efficacy dropped to 61%
across the four materials when physical removal was decoupled from the efficacy calculation.
Under the same test conditions, the 1:4 diluted Dahlgren Decon™ decontaminant demonstrated
efficacies of 96% across the four materials attributable to a combination of physical removal and
chemical decontamination and the average percent efficacy dropped to 95% across the four
materials when physical removal was decoupled from the efficacy calculation.
Generally, the 1:4 diluted Dahlgren Decon™ demonstrated noticeable higher average
decontamination efficacies than the pH 5 modified surfactant bleach decontaminant after the 5-
min dwell time.
5.3 Clustering of Fentanyl Powder
In many of the decontamination tests, agglomerated fentanyl was observed visually on surfaces
following the application and dwell time of the decontaminant. This clustering or clumping of
fentanyl on the surface results in a slower mass transfer rate between the decontaminant and
fentanyl. Hence, higher amounts were recovered on occasion even in the presence of an
otherwise effective decontaminant. Such behavior may also occur in actual remediation efforts
and should be watched for.
62
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REFERENCES
1. Oudejans L., D. See, C. Dodds, M. Corlew, and M. Magnuson, Decontamination Options
for Indoor Surfaces Contaminated with Realistic Fentanyl Preparations. Journal of
Environmental Management 297 (2021), 113327.
2. Doerflein, J. Case Study: A Fentanyl Incident.
https://www.epa.gov/sites/production/files/2020-02/documents/doerflein fentanyl.pdf Last
accessed April 30, 2021.
3. Fisher, R. A. (1942). The Design of Experiments, 3rd Edition, Edinburgh: Oliver & Boyd.
4. Miller, R. G., Jr. (1981). Simultaneous Statistical Inference. New York: Springer-Verlag.
5. Tukey, J. W. (1953). "The Problem of Multiple Comparisons." In Multiple Comparisons,
1948-1983, edited by H. I. Braun, vol. 8 of The Collected Works of John W. Tukey
(published 1994), 1-300. London: Chapman & Hall.
6. SW-846 Method 8000D, Determinative Chromatographic Separations. Revision 5, March
2018, Final Update V to the Third Edition of the Test Methods for Evaluating Solid Waste,
Physical/Chemical Methods, EPA Publication SW-846.
63
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Attachment A - Fentanyl Certificate of Analysis
64
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Cayman Chemical Co.
»1HU t. blteworth Kd
Ann Arbor, Ml 4B108
www.caymanchem cam
Contact Us:
1-UtHJ-3tS4-9B9/ or(/M)!iM-D3l>
Fa* (734)971-3640
Ems': crmqualityigjesyinanehem com
Fcr R&D purposes yvy- «er t»enaeatwl»mai>orai\tnml use.
CERTIFICATE OF ANALYSIS
Fenlanyl (hydrochloride]
N-phenyl-N-(l-(2-pheny]ethy))-4-piperidiny]1-prapaiiamlde, monohydrochluride
Item#: 14719
Batch #: 0530926
CAS Registry (lumber: 1443-54-5
Expiry Date: 01JUN2023 (valid from date of certification)
Description: neat solid
Storage and Handling: Store at -20'C. Warm to room
temperature prior to opening.
Safety: Poison
HCI
Chemical f ormula
CnlbflNiU • HCI
Formula Weight
372,90 amu
UVJW
205 nm
ISOilEC 170IS
•AT-1773
HOGuldt 34
IAR-1774
AKAB
accredited
ACCREDITED
TF^
Quality information
Qualifier
Method
Limit
Result
Meets
Specification
Appearance
Visual inspection
White / off-while
solid
Off-white solid
y
Chromatographic
Puritv, HPLC
Cayman Method TSTSD132
298.00%
99.59% ± 0.18%
V
Identity, LCMS
Cayman Method TST SD13, +ESI
337.2 ±0,5 amu
337.4 amu
¥
Identity, CCMS
Cayman Method 1ST SD12
Conforms
Conforms
V
FTIR
U5F-=B51> (diamond ATR)
Conforms
Conforms
Y
~Identity, NMR
iH NMR
Conforms
Conforms
Y
Loss on Drying;
Cayman Method TST SD24
£10.00%
0.38% ±0.49%
Y
Residue on Ignition
Cayman Method TST SD06
£3.00%
<0.10% ± 0.22%
Y
'NMR is provided as supp
eme "ta 1 info but is not within scope of ISO accreditation
Property values are traceable to SI units through an unbroken chain of measurements.
Measurement Uncertainty
All measurement uncertainties are expressed as expanded uncertainties in accordance with ISO 17Q25 and Guide 34 at the approximate 95%
confidence interval using a coverage factor of k=2.
Cayman Chemical certifies that this standard meets the specifications stated in this certificate and warrants this product to
meet the stated acceptance criteria through the expiration date when stored unopened as recommended.
Approval:
Title: Cayman Chemical ISO Quality Manager Certification Date: 01JUN2018
Page 1 of 5
Certificate # 14719-0530926-01
65
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Cayman Chemi cat Co. Contact Us: fir R&D Durroses any: Net Intended tar human or antra' use
1130 E. Blsworlh Rd 1-8Q0-364-&887 or (734)971-3335
Ann Arbor, M1481GB Fax: (734)971 -3640
'AVfW.caymanchenn.com Email: crrrqualrty@caymanchem.com
Supplemental Data
HPLC-UV
DAD1 A. Sig=210.8 Ref=450.1 B (C:\CAYMANV.. FENTANYL HCL\D530926 PURITY 2t>
Norm
800-
600-
400-
200-
o-
-200-
-400-
? 5
5^
Ir
\h$i-
Conditions
4.6x150mm, 5jjrn Luna Ptienyi-Hexyl column
A: 0.1 % TFA in H:0
B: methanol
D-10 minutes 20-05% B, 10-13 minutes 95% B,
13.1-20 m^Jtes 20% B
1ml/min
30 cofemrn temp
UV moniorec at 210rm
r s
in
\7 5
GC-MS
AtmiMno
W3QIK
iwm:
220001'
1KMH1G
iw:n>.
1ZOOOC
900Q9
flaott
*ooo:
20000
mil—•
Aw»9* rf 11 •'Stl la 11 rr»n 14Tl(Mi£d£>a» Urfrtn rmi (-*
hU -
, . , ,
fid ,'fi ad uti ion avii lin
[l ttaip iKflj
iui ifc isi isw /in 2iti i'ii: vim mc IL ¦/til j fii
Conditions
Agilent 6390 GO
5673 MSD
30m x 0.32mm, O.fum
Rtx-5MS column
He carrier gas;
Row: 2mL'min
Wet temp: 300*C
15:1 split
Oven Program;
50'C hold for 1 minute,
ramp to 300'C at
30eC / minyte, hoW at
30D'C to 15 minutes
Transfer line temp:
300'C
70ev El MS
40-600 ntfz
Apex spectrum -
background
(1 minute w.ndow in
front of peak)
T-jne File: stune
, ky-i ?*< J?9 ^;7T|
Page 2 of 5
Certificate # 14719-0530926-01
66
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Cayman Chemical Co. Contact Us: Fcr R&D ixivoses cnty. t&t IrtiencttltorHuman or anim&'use.
1180 E. El!5worth Rd 1 -BD0-364-9B97 or <734)971-3335
Ann Arbor, M148108 Fax: (734)971 -3840
v»v«w. caymanchem.com Email: crmquality@caymanchem.com
FUR
\
-------
Cayman Chemical Co.
1180 E. Ellsworth Rd
Ann Arbor, Ml 481DB
vww.cayrnanchem.com
Contact Us:
1 -BQ0-364-&B97 or <734)971-3335
Fax: (734)971-3640
Email: crmquality@caymanchem.com
Far R&D djto505 •amy wcr .Tfe/tfea fir ALmar ar aniTA' use.
NMR ''not within scope of ISO 17025/Guide 34 accreditation
aArwAa i f,Mj, j}If,TO i Tw«6i !j+2 Wr»S28_'>f.T
IriHIMi K1EC-S-; Or Vfcfc
HkKDCt^llKIWU in
gbhvnl wwaam. 1 liMMkrr.WMi'C' mam
!I1 NMR |V1UI«,Wi:t.-iJJl4«M MJ#f J S-Thim, 111 I 7.2?.4 |n. Till.
4JMlS(m. Ill), .i.tt- Ibr d, 211. J-12.3 ILrL JJ-3J Im. 211K i.l J.2 Im,
lli|,.V4-V6irnJil>, i U <1wJ, 111, J- *4.1 M** l.«Mq, III, J T.4 iVi.
1 73^. 2IL./ J.S. I3.Z If*). 3IT. f-lJbIfag
Conditsns
Varian bwva 400MHz NMR
64 scans
tis5*
V'-
»«*
ti
0.«6
u
'(J®
Charazal filth ]Bpn,l
Homogeneityr
Homogeneity was assessed by visual inspection and replicate purity analyses. The recommended minimum quantity for use is
2.0 |lg» Quantities below this have not been evaluated.
Shoit Term Stability? Summary
No decrease in purity was observed at ambient conditions or 60"C after two weeks. This data supports shipping of this
product at ambient temperature.
Long Term Stability?
Long term stability data predicts 5 years stability at the -20?C storage temperature.
Page 4 of 5
Certificate # 14719-0530926-01
68
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Cayman Chemical Co.
1180 E Ellsworth Rd
Ann Arbor, Ml 48108
•Aww.cayma nchem.com
Contact Us:
1 -803-364-8 B57 or (734)971-3335
Fax: 1734)971-3640
Email: crrrvqualityig1 caymanchem.com
Fcr R&D purposes anty Net intended fzr .iLAmar? ar antoBf use.
Revision History
Revision #
Date Reason for Revision
01
01JUN2018 Initial version
Dfariatmre
Material Safety Data
TTse: masts! should be corattFEd ftsardciE Jrti ffonrsaon 1d the Gcrrtrary beccnes ava atie. Do not ingest swaisw, x'maee. Do not get r eyes, on sWr% cran actfftng Wash BvHugfty after
hard r$. T-fc rite-acor conaa-s bid not a 3" the i-forrufcon required tr tfte sate arc! proper use cf tis -ratera. 5Cere use rertei# me corciete '-•arena Safety Dar-s £hee", often has oee*i
xt. vaerrai b >caue care aid ska. unus, in rc event ¦aHI Caymai have any ¦z&bjatcri or UUBy, •Mvrer ri tort iiTciKSng ne# gerce;> v r contract, tf any d*Bd,
Indfree, rodents cr conscqjenaa damages, e«n ff Cayrnar Is rrfor-ed aoox oossfce erasence.
The trtatcr ofbbfty does roc acptj- :n re case of ntntcrai acrs or neg*gence or* Cayman. it: dwetars or its em»cy«s.
Bjyer'z odusfm rarvKfc and Cayman's s»e iUURy heretnder shall be I rrted b a refjnd of tw sjfase pice, or si Cayman1* option the repaccTerr. at recast» Buyer, of a I PH&ertal fa! ctoes ret
meet o\r spec?fcafl:r.
Said rcTirfl or repacerre*t: Is conAcred on Buyers vino wrtten notice to Cajmar wlftn tftfrtrlSQ oay- after anM or the Tiatsra; at ts SKsnaton. Fa ire cf Buyer x 7»? sad notee Hlh*i Hitjr i3D;i
oa/s shafi constat a wafts- of Buyer 3* a cafins heetr>der ulth respect to said natertal
FvLrtherdetafe please refer to otrWarrarty and Lmtaflxs DfReniedji- ocated or aurMdisteand n :
-------
Attachment B - Environmental Data
70
-------
Table Bl. Environmental Conditions in Experimental Chamber with Spray Setup
Test Description
Temperature Range
(°C)
Relative Humidity
(%)
Quench Method development (no spray)
19.8-21.1
22-27
Meth Remover® decontamination (60 min)
19.9-22.0
56-94
ZEP® decontamination (60 min)
19.7-22.2
60-94
Dahlgren Decon™ decontamination (60 + 60 min)
20.1-22.4
57-76
pH 5 modified surfactant bleach decontamination (60 + 60 min)
19.6-22.1
60-91
Diluted Dahlgren Decon™ decontamination (various time
points up to 15 min)
19.4-21.6
63-92
Diluted Dahlgren Decon™ decontamination (5 min)
20.9-22.7
56-99
pH 5 modified surfactant bleach decontamination (5 min)
19.3-21.5
64-97
71
-------
Attachment C - Spray Characterization Data
72
-------
Table CI. Decontaniinant Spray Delivery Mass per Position, Meth Remover®
Position
Rep 1
Rep 2
Sprayed Decontaminant Weight (g)
Rep 3 Average St Dev % RSD
Percent of Target
1
1.30
1.31
1.22
1.28
0.049
3.9%
110%
2
1.23
1.31
1.11
1.22
0.101
8.3%
105%
3
1.23
1.35
1.07
1.22
0.140
11.5%
105%
4
1.30
1.35
1.01 A
1.22
0.184
15.0%
105%
5
1.37
1.35
1.27
1.33
0.053
4.0%
114%
6
1.38
1.28
1.31
1.32
0.051
3.9%
114%
7
1.21
1.36
1.28
1.28
0.075
5.8%
110%
8
1.38
1.32
1.24
1.31
0.070
5.3%
113%
9
1.22
1.37
1.31
1.30
0.075
5.8%
112%
10
1.24
1.38
1.33
1.32
0.071
5.4%
113%
11
1.26
1.24
1.14
1.21
0.064
5.3%
104%
12
1.23
1.26
1.32
1.27
0.046
3.6%
109%
13
1.32
1.36
1.39
1.36
0.035
2.6%
117%
14
1.28
1.39
1.35
1.34
0.056
4.2%
115%
15
1.19
1.35
1.35
1.30
0.092
7.1%
111%
16
1.24
1.32
1.36
1.31
0.061
4.7%
112%
A Position 4, rep 3 small amount of decontaminant spilled before weighing
Table C2. Decontaminant Spray Delivery Mass per Position, ZEP®
Position
Rep 1
Rep 2
Sprayed Decontaminant Weight (g)
Rep 3 Average St Dev % RSD
Percent of Target
1
1.18
1.07
1.13
1.13
0.055
4.9%
97%
2
1.19
1.13
1.11
1.14
0.042
3.6%
98%
3
1.28
1.19
1.07
1.18
0.105
8.9%
102%
4
1.30
1.21
1.17
1.23
0.067
5.4%
106%
5
1.26
1.20
1.24
1.23
0.031
2.5%
106%
6
1.36
1.24
1.29
1.30
0.060
4.6%
112%
7
1.35
1.26
1.24
1.28
0.059
4.6%
111%
8
1.28
1.13
1.09
1.17
0.100
8.6%
100%
9
1.00
0.96
0.99
0.98
0.021
2.1%
85%
10
0.98
0.97
0.98
0.98
0.006
0.6%
84%
11
0.95
0.97
0.98
0.97
0.015
1.6%
83%
12
0.99
1.00
0.99
0.99
0.006
0.6%
86%
13
1.01
0.98
0.99
0.99
0.015
1.5%
86%
14
1.02
1.00
1.02
1.01
0.012
1.1%
87%
15
1.05
1.01
0.97
1.01
0.040
4.0%
87%
16
1.03
0.99
0.94 A
0.99
0.045
4.6%
85%
A Position 16, rep 3 small amount of decontaminant spilled before weighing.
-------
Table C3. Decontaminant Spray Delivery Mass per Position, pH 5 Modified Surfactant
Bleach
Position
Rep 1
Rep 2
Sprayed Decontaminant Weight (g)
Rep 3 Average St Dev % RSD
Percent of Target
1
1.08
1.19
1.08
1.12
0.064
5.7%
99%
2
1.12
1.13
1.20
1.15
0.044
3.8%
102%
3
1.16
1.16
1.12
1.15
0.023
2.0%
101%
4
1.18
1.16
1.11
1.15
0.036
3.1%
102%
5
1.19
LostA
1.14
1.17
0.035
3.0%
103%
6
1.17
1.13
1.10
1.13
0.035
3.1%
100%
7
1.15
1.16
1.10
1.14
0.032
2.8%
100%
8
1.11
1.09
1.11
1.10
0.012
1.0%
97%
9
1.05
1.05
1.07
1.06
0.012
1.1%
93%
10
1.12
1.08
1.06
1.09
0.031
2.8%
96%
11
1.10
1.09
1.09
1.09
0.006
0.5%
97%
12
1.14
1.07
1.07
1.09
0.040
3.7%
97%
13
1.09
1.09
1.11
1.10
0.012
1.1%
97%
14
1.13
1.21
1.19
1.18
0.042
3.5%
104%
15
1.10
1.17
1.14
1.14
0.035
3.1%
100%
16
1.11
1.15
1.18
1.15
0.035
3.1%
101%
A Position 5, rep 2 sample lost (spilled).
Table C4. Decontaminant Spray Delivery Mass per Position, Dahlgren Decon™
Position
Rep 1
Rep 2
Sprayed Decontaminant Weight (g)
Rep 3 Average St Dev % RSD
Percent of Target
1
1.34
1.11
1.54
1.33
0.215
16.2%
104%
2
1.22
1.15
1.39
1.25
0.123
9.8%
98%
3
1.20
1.08
1.24
1.17
0.083
7.1%
92%
4
1.19
1.30
1.20
1.23
0.061
4.9%
96%
5
1.28
1.18
1.30
1.25
0.064
5.1%
98%
6
1.42
1.33
1.13
1.29
0.148
11.5%
101%
7
1.36
1.38
1.23
1.32
0.081
6.2%
103%
8
1.25
1.53
1.28
1.35
0.154
11.4%
106%
9
1.50
1.25
1.34
1.36
0.127
9.3%
107%
10
1.47
1.28
1.66
1.47
0.190
12.9%
115%
11
1.45
1.27
1.61
1.44
0.170
11.8%
113%
12
1.32
1.15
1.51
1.33
0.180
13.6%
104%
13
1.38
1.24
1.43
1.35
0.098
7.3%
106%
14
1.36
1.20
1.20
1.25
0.092
7.4%
98%
15
1.40
1.14
1.28
1.27
0.130
10.2%
100%
16
1.31
1.31
1.30
1.31
0.006
0.4%
102%
-------
Table C5. Decontaminant Spray Delivery Mass per Position, Diluted Dahlgren Decon™
Position
Rep 1
Rep 2
Sprayed Decontaminant Weight (g)
Rep 3 Average St Dev % RSD
Percent of Target
1
1.31
1.18
1.23
1.24
0.066
5.3%
107%
2
1.26
1.18
1.21
1.22
0.040
3.3%
105%
3
1.20
1.17
1.22
1.20
0.025
2.1%
103%
4
1.18
1.24
1.15
1.19
0.046
3.9%
102%
5
1.25
1.18
1.17
1.20
0.044
3.6%
103%
6
1.25
1.17
1.15
1.19
0.053
4.4%
102%
7
1.17
1.17
1.01
1.12
0.092
8.3%
96%
8
1.19
1.14
1.14
1.16
0.029
2.5%
100%
9
1.13
1.07
1.11
1.10
0.031
2.8%
95%
10
1.20
1.12
1.12
1.15
0.046
4.0%
99%
11
1.12
1.07
1.01
1.07
0.055
5.2%
92%
12
1.02
0.99
0.98
1.00
0.021
2.1%
86%
13
1.06
1.07
1.09
1.07
0.015
1.4%
92%
14
1.16
1.02
1.10
1.09
0.070
6.4%
94%
15
1.15
1.02
1.06
1.08
0.067
6.2%
93%
16
1.15
1.11
1.10
1.12
0.026
2.4%
96%
-------
Attachment D - Average Mass Recovery and Decontamination Efficacy Data
76
-------
Table Dl. Average Mass Recovery, Meth Remover®
Average Recovery
Decontaminant
Material
Sample
Coupon
Runoff
Total Mass (Coupon + Runoff)
Description
Mass
St. Dev.
RSD
% of Total
Mass
St. Dev.
RSD
% of Total
Mass
St. Dev.
RSD
% Recovery
(tig)
(tig)
(%)
Mass
(fig)
(tig)
(%)
Mass
(fig)
(fig)
(%)
vs Pos Avg
Painted
Positive Controls
1077
189
18%
-
-
-
-
-
-
-
-
drywall
Test Coupons
677 A
169
25%
81%
149
29
19%
19%
827
143
17%
77%
Coated
Positive Controls
1379
254
18%
-
-
-
-
-
-
-
-
-
Meth
steel
Test Coupons
346 A
124
36%
58%
235
31
13%
42%
582
139
24%
42%
Remover®
Laminate
Positive Controls
1180
144
12%
-
-
-
-
-
-
-
-
-
Test Coupons
438 A
102
23%
54%
381
105
28%
46%
818
105
13%
69%
Wood
Positive Controls
1251
252
20%
-
-
-
-
-
-
-
-
-
Test Coupons
337
54
16%
61%
211
44
21%
39%
548
31
5.6%
44%
A Solid material observed on replicate coupon surfaces at time of extraction.
Table D2. Average Mass Recovery, ZEP®
Average Recovery
Decontaminant
Material
Sample
Coupon
Runoff
Total Mass (Coupon + Runoff)
Description
Mass
St. Dev.
RSD
% of Total
Mass
St. Dev.
RSD
% of T otal
Mass
St. Dev.
RSD
% Recovery
(fig)
(fig)
(%)
Mass
(fig)
(fig)
(%)
Mass
(fig)
(fig)
(%)
vs Pos Avg
Painted
Positive Controls
1235
5.4
0.44%
-
-
-
-
-
-
-
-
drywall
Test Coupons
529 A
241
46%
50%
513
150
29%
50%
1042
183
18%
84%
Coated
Positive Controls
1375
143
10%
-
-
-
-
-
-
-
-
-
ZEP®
steel
Test Coupons
306 A
166
54%
30%
694
198
29%
70%
999
196
20%
73%
Laminate
Positive Controls
1351
195
14%
-
-
-
-
-
-
-
-
-
Test Coupons
424 A
54
13%
36%
744
120
16%
64%
1168
172
15%
oN
\Q
00
Wood
Positive Controls
1266
59
4.7%
-
-
-
-
-
-
-
-
-
Test Coupons
545 A
139
25%
79%
138
67
49%
21%
683
79
12%
54%
A Solid material observed on replicate coupon surfaces at time of extraction.
77
-------
Table D3. Decontamination Efficacy Testing, Average Percent Efficacy
Decontaminant
Material
Avg % Efficacy ± SD
[physical removal and chemical
decontamination] (%) A
Avg % Efficacy ± SD
[chemical decontamination only]
(%)B
Meth Remover®
Painted drywall
37 ± 19
23 ± 19
Laminate
75 ± 10
58 ± 13
Coated steel
63 ± 10
31 ±12
Wood
73 ±7
56 ±9
ZEP®
Painted drywall
57 ±20
16 ± 15
Laminate
78 ± 12
27 ± 16
Coated steel
69 ±6
14 ± 18
Wood
57± 11
46 ±7
A Avg test coupon recovery vs avg pos control recovery. Combined efficacy of physical removal and chemical degradation.
B Avg test coupon recovery plus avg runoff recovery vs avg pos control recovery. Efficacy of chemical degradation only.
Table D4. Average Mass Recovery, Dahlgren Decon™ — Double Application
Average Recoveiy
Sample
Description
Coupon
Runoff
Total Mass (Coupon + Runoff)
Decontaminant
Material
Mass
St. Dev.
RSD
% of Total
Mass
St. Dev.
RSD
% of Total
Mass
St. Dev.
RSD
%
Recovery vs
Pos Avg
(tig)
(tig)
(%)
Mass
(fig)
(tig)
(%)
Mass
(fig)
(tig)
(%)
Painted
Positive Controls
1371
108
7.3%
-
-
-
-
-
-
-
-
-
drywall
Test Coupons
0.10
0.09
83%
9.2%
1.5
0.7
46%
91%
1.6
0.62
38%
0.12%
Coated steel
Positive Controls
1688
329 A
19%
-
-
-
-
-
-
-
-
-
Dahlgren
Test Coupons
0.04
0.02
53%
1.7%
1.9
0.49
25%
98%
2.0
0.5
26%
0.12%
Decon™
Wood
Positive Controls
1268
65
5.1%
-
-
-
-
-
-
-
-
-
Test Coupons
1.2
0.83
72%
44%
1.2
0.40
34%
56%
2.3
1.1
47%
0.18%
Wood with
Positive Controls
1406
53 A'B
3.7%
-
-
-
-
-
-
-
-
-
ascorbic acid
Test Coupons
42
40
95%
89%
2.4
1.1
46%
11%
44
39
89%
3.1%
A Solid material observed on replicate coupon surfaces following 1st 60-min application.
B Solid material observed on replicate coupon surfaces at time of extraction.
78
-------
Table D5. Average Mass Recovery, pH 5 Modified Surfactant Bleach — Double Application
Average Recovery
Decontaminant
Material
Sample
Coupon
Runoff
Total Mass (Coupon + Runoff)
Description
Mass
St. Dev.
RSD
% of Total
Mass
St. Dev.
RSD
% of Total
Mass
St. Dev.
RSD
% Recovery
(tig)
(tig)
(%)
Mass
(fig)
(tig)
(%)
Mass
(fig)
(fig)
(%)
vs Pos Avg
Painted
Positive Controls
1208
8
0.66%
-
-
-
-
-
-
-
-
drywall
Test Coupons
6.1 AB
3.1
50%
15%
37
15
41%
85%
43
16
39%
3.5%
pH 5 modified
surfactant
bleach
Coated steel
Positive Controls
1208
13
1.1%
-
-
-
-
-
-
-
-
-
Test Coupons
42 A'B
60
145%
24%
67
49
73%
76%
109
110
101%
9.0%
Wood
Positive Controls
1204
81
6.7%
-
-
-
-
-
-
-
-
-
Test Coupons
139 AB
183
132%
63%
52
45
85%
37%
191
227
119%
16%
Wood with
Positive Controls
1274
59
4.7%
-
-
-
-
-
-
-
-
-
ascorbic acid
Test Coupons
148 A B
117
12%
55%
106
40
37%
45%
254
132
52%
20%
A Solid material observed on replicate coupon surfaces following 1st 60-min application.
B Solid material observed on replicate coupon surfaces at time of extraction.
Table D6. Decontamination Efficacy Testing, Average Percent Efficacy
Decontaminant
Material
Avg % Efficacy ± SD
[physical removal and chemical
decontamination] (%) A
Avg % Efficacy ± SD
[chemical decontamination only]
(%)B
Dahlgren Decon™
Painted drywall
99.992 ± 0.006
99.88 ±0.05
Coated steel
99.998 ±0.001
99.88 ±0.04
Wood
99.91 ±0.07
99.82 ±0.09
Wood with ascorbic acid
97 ±3
97 ±3
pH 5 modified
surfactant bleach
Painted drywall
99.5 ±0.3
96 ± 1
Coated steel
97 ±5
91 ±9
Wood
88 ± 15
84 ± 19
Wood with ascorbic acid
88 ±9
80 ± 10
A Avg test coupon recovery vs avg pos control recovery. Combined efficacy of physical removal and chemical degradation.
B Avg test coupon recovery plus avg runoff recovery vs avg pos control recovery. Efficacy of chemical degradation only.
79
-------
Table D 7. Average Mass Recovery, Dahlgren Decon™
Average Recovery
Sample
Description
Coupon
Runoff
Total Mass (Coupon + Runoff)
Decontaminant
Material
Mass
St. Dev.
RSD
% of Total
Mass
St. Dev.
RSD
% of Total
Mass
St. Dev.
RSD
%
Recovery vs
Pos Avg
(tig)
(tig)
(%)
Mass
(fig)
(tig)
(%)
Mass
(fig)
(tig)
(%)
Saranex®
Positive Controls
1021
52
5.1%
-
-
-
-
-
-
-
-
-
Test Coupons
224 A
261
116%
41%
239
in
46%
59%
463
171
37%
45%
pH 5 modified
surfactant
bleach
HazMat suit
Positive Controls
1069
57
5.3%
-
-
-
-
-
-
-
-
-
Test Coupons
151 A
56
37%
35%
272
47
17%
65%
423
47
11%
40%
Bunker gear
Positive Controls
1135
136
12%
-
-
-
-
-
-
-
-
-
Test Coupons
118 A
100
85%
29%
273
57
21%
24%
391
44
11%
34%
Neoprene
Positive Controls
1145
159
14%
-
-
-
-
-
-
-
-
-
Test Coupons
119 A
151
128%
26%
320
131
41%
74%
438
35
7.9%
38%
A Solid material observed on replicate coupon surfaces at time of extraction.
Table D8. Average Mass Recovery, pH 5 Modified Surfactant Bleach
Average Recovery
Decontaminant
Material
Sample
Coupon
Runoff
Total Mass (Coupon + Runoff)
Description
Mass
St. Dev.
RSD
% of Total
Mass
St. Dev.
RSD
% of Total
Mass
St. Dev.
RSD
% Recovery
(tig)
(tig)
(%)
Mass
(fig)
(tig)
(%)
Mass
(fig)
(tig)
(%)
vs Pos Avg
Saranex®
Positive Controls
1182
322
27%
-
-
-
-
-
-
-
-
-
Test Coupons
2.3
2.5
109%
18%
15
15
99%
82%
18
18
99%
1.5%
Diluted
Dahlgren
Decon™
HazMat suit
Positive Controls
992
52
5.2%
-
-
-
-
-
-
-
-
-
Test Coupons
103 A
72
69%
90%
8.0
10
126%
9.6%
Ill
74
66%
11%
Bunker gear
Positive Controls
917
36
3.9%
-
-
-
-
-
-
-
-
-
Test Coupons
28 A
8.9
32%
87%
4.9
4.4
91%
13%
33
12
38%
3.6%
Neoprene
Positive Controls
822
105
13%
-
-
-
-
-
-
-
-
-
Test Coupons
13 A
6.8
52%
81%
3.4
5.0
149%
19%
16
6.4
39%
2.0%
A Solid material observed on replicate coupon surfaces at time of extraction.
80
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Table D9. Decontamination Efficacy Testing, Average Percent Efficacy
Decontaminant
Material
Avg % Efficacy ± SD
[physical removal and chemical
decontamination] (%)A
Avg % Efficacy ± SD
[chemical decontamination
only] (%) B
pH 5 modified
surfactant bleach
Saranex®
78 ±25
55 ± 17
HazMat suit
86 ±5
60 ±5
Bunker gear
90 ±9
66 ±6
Neoprene
90 ± 13
62 ±6
Diluted Dahlgren
Decon™
Saranex®
99.8 ±0.2
98 ±2
HazMat suit
90 ±7
89 ±7
Bunker gear
97 ± 1
96 ± 1
Neoprene
98 ± 1
98 ± 1
A Avg test coupon recovery vs avg pos control recovery. Combined efficacy of physical removal and chemical degradation.
B Avg test coupon recovery plus avg runoff recovery vs avg pos control recovery. Efficacy of chemical degradation only.
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82
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vvEPA
United States
Environmental Protection
Agency
PRESORTED STANDARD
POSTAGE & FEES PAID
EPA
PERMIT NO. G-35
Office of Research and Development (8101R)
Washington, DC 20460
Official Business
Penalty for Private Use
$300
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