oEPA
EPA/600/R-15/188 I September 2015 I www2.epa.gov/research
United States
Environmental Protection
Agency
Operational Testing of Sporicidal
Wipes for Decontamination of
Surfaces Contaminated with
Bacillus anthracis Surrogate Spores
ASSESSMENT AND EVALUATION REPORT
Office of Research and Development
National Homeland Security Research Center
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f/EPA
Assessment and Evaluation Report
Operational Testing of Sporicidal Wipes for Decontamination
of Surfaces Contaminated with Bacillus anthracis Surrogate
Spores
1
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Operational Testing of Sporicidal Wipes for Decontamination
of Surfaces Contaminated with Bacillus anthracis Surrogate
Spores
Assessment and Evaluation Report
National Homeland Security Research Center
Office of Research and Development
U.S. Environmental Protection Agency
Research Triangle Park, NC 27711
2
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Disclaimer
The U.S. Environmental Protection Agency through its Office of Research and Development funded and
managed the research described here under contract EP-C-09-027, Work Assignments 5-73 and 6-73, with
ARCADIS U.S., Inc. 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. Mention of
trade names, products, or services does not convey official EPA approval, endorsement, or recommendation
Questions concerning this document or its application should be addressed to the principal investigator:
Lukas Oudejans, Ph.D.
U.S. Environmental Protection Agency
Office of Research and Development
National Homeland Security Research Center
109 T.W. Alexander Drive (Mail Drop: E343-06)
Research Triangle Park, NC 27711
Phone: (919) 541-2973
Email: Oudejans.Lukas@epa.gov
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Acknowledgments
This effort was directed by the principal investigator (PI) from the Office of Research and Development's
(ORD) National Homeland Research Center (NHSRC), to address critical capability or knowledge gaps
identified by NHSRC's Program to Align Research and Technology with the Needs of Environmental
Response (PARTNER).
Project Team:
Lukas Oudejans, Ph.D. [Principal Investigator]
NHSRC, ORD, US EPA
Sang Don Lee, Ph.D.
NHSRC, ORD, US EPA
M. Worth Calfee, Ph.D.
NHSRC, ORD, US EPA
Leroy Mickelsen
CMAD, OEM, US EPA
This effort was completed under U.S. EPA contract #EP-C-09-027 with ARCADIS-US, Inc. The support and
efforts provided by ARCADIS-US, Inc. are gratefully acknowledged.
Also acknowledged are;
Ramona Sherman and Eletha Brady-Roberts (Quality Assurance)
NHSRC, ORD, US EPA
Cincinnati, OH 45220
Joan Bursey (Editorial Review)
NHSRC, ORD, US EPA
Research Triangle Park, NC 27711
The peer reviewers of this report are also acknowledged for their input to this product:
Joseph Wood, ORD/NHSRC
Shannon Serre, OEM/CMAD
Benjamin Franco, Region 3
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Table of Contents
Disclaimer iii
Acknowledgments iv
List of Tables viii
List of Figures xi
Acronyms and Abbreviations xii
Executive Summary xiv
1 Introduction 1
1.1 Background 1
1.2 Project Objectives and Process 1
2 Experimental Approach 2
2.1 Project Description 2
2.1.1 PHASE 1: Operational Aspects of Decontamination by Sporicidal Wipes 2
2.1.2 PHASE 2: Evaluation of Efficacy of Hypochlorite Wipes 3
2.2 Testing Program Description 3
2.2.1 PHASE 1: Operational Aspects of Decontamination by Sporicidal Wipes 3
2.2.1.1 Task 1 - Determination of Effective Surface Coverage of Sporicidal Wipes 4
2.2.1.2 Task 2 - Surface Retention Times of Wipe Decontaminant Liquid 5
2.2.1.3 Task 3 - Impact of Wiping Pressure 5
2.2.2 PHASE 2: Evaluation of Efficacy of Hypochlorite Wipes 6
2.3 Definitions of Effectiveness 9
3 Material and Methods 11
3.1 Test Coupon Preparation and Sterilization 11
3.2 Bacillus Spore Preparation 11
3.3 Coupon Inoculation Procedures 12
3.4 Decontamination Procedure 13
3.5 PHASE 1 Measurement Protocol 14
3.5.1 Sampling Approach 14
3.5.2 Gravimetric Measurements 14
3.5.3 Method Verification 15
3.6 PHASE 2 Measurement Protocol 15
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3.6.1 Sterile Handling ofWipes 15
3.6.2 Sampling Site Environmental Conditions 15
3.6.3 Wipe Sampling Procedures 16
3.6.4 Swab Sampling 19
3.6.5 Phase 1 Sample Identification 19
3.6.6 Phase 2 Sample Identification 20
3.6.7 Sample Frequency 21
3.6.8 Prevention of Cross Contamination of Samples during Sampling 23
3.6.9 Collecting Representative Samples 23
3.6.10 Sample Storage and Preservation 24
3.6.11 Sample Holding Time 24
3.6.12 Sample Chain of Custody 24
3.6.13 Sample Archiving 25
3.6.14 Microbiological Analysis 25
4 Results and Discussion 26
4.1 PHASE 1: Operational Aspects of Decontamination by Sporicidal Wipes Results 26
4.1.1 Determination of Effective Surface Coverage of Sporicidal Wipes 26
4.1.2 Surface Retention Times ofWipe Decontaminant Liquid Results 28
4.1.3 Impact of Wiping Pressure 30
4.2 PHASE 2: Evaluation of Efficacy of Hypochlorite Wipes Results 31
5 Quality Assurance and Quality Control 39
5.1 Criteria for Critical Measurements/Parameters 39
5.2 Quality Control Checks 40
5.2.1 Integrity of Samples and Supplies 40
5.2.2 NHRSC Biolab Control Checks 41
5.3 QA/QC Sample Acceptance Criteria 41
5.3.1 QA/QC Test Results Validation 43
5.3.2 Sample Hold Time 44
5.4 Instrument Calibrations 47
5.5 QA Assessments and Response Actions 48
5.6 Data Reduction 49
5.7 Data Reporting 49
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5.8 QAPP Amendments and Deviations 49
5.8.1 Formal Amendments 49
5.8.2 QAPP Deviations 50
6 SUMMARY 51
7 REFERENCES 53
APPENDIX A - PHASE 1: Operational Aspects of Decontamination by Sporicidal Wipes 54
Test Results for Task 1: Determination of Effective Surface Coverage of Sporicidal Wipes 54
Test Results for Task 2: Surface Retention Times of Wipe Decontaminant Liquid 70
Test Results for Task 3: Impact of Wiping Pressure 78
APPENDIX B - PHASE 2: Evaluation of Efficacy of Hypochlorite Wipes 86
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List of Tables
Table 2-1. Sporicidal Wipe Information 3
Table 2-2. Task 1 Test Matrix Conducted for Each Type of Sporicidal Wipe 4
Table 2-3. Task 2 Test Matrix per Sporicidal Wipe Type 5
Table 2-4. Task 3 Test Matrix per Sporicidal Wipe Type 6
Table 2-5. Decontamination Test Matrix for Wiping Surfaces with Sporicidal Wipes 8
Table 3-1. Sample Coding for Phase 1 20
Table 3-2: Sample Coding for Phase 2 21
Table 3-3. Wiping Efficacy - Sample Frequency 22
Table 4-1. Decontamination Results for Glass Material Type 33
Table 4-2. Decontamination Results for Painted Drywall Material Type 34
Table 5-1. Critical Measurement Criteria 40
Table 5-2. QA/QC Sample Acceptance Criteria 42
Table 5-3. QA/QC Test Results 43
Table 5-4. Test Matrix (Test Samples) 45
Table 5-5. Test Matrix - Hold Time Control Samples 46
Table 5-6. Sample Hold Time Test Results for Decontamination Efficacy Tests 47
Table 5-7. Instrument Calibration Frequency 48
Table A-1-1-1: Task 1 Results Clorox Healthcare® Bleach Germicidal Wipe/Vinyl Flooring 54
Table A-1-1-2: Task 1 Results Clorox Healthcare® Bleach Germicidal Wipe/Glass 55
Table A-1-1-3: Task 1 Results Clorox Healthcare® Bleach Germicidal Wipe/Melamine 56
Table A-1-1-4: Task 1 Results Clorox Healthcare® Bleach Germicidal Wipe/Painted Drywall 57
Table A-1-2-1: Task 1 Results Hype-Wipe® Bleach Towelette/Vinyl Flooring 58
Table A-1-2-2: Task 1 Results Hype-Wipe® Bleach Towelette/Glass 59
Table A-1-2-3: Task 1 Results Hype-Wipe® Bleach Towelette/Melamine 60
Table A-1-2-4: Task 1 Results Hype-Wipe® Bleach Towelette/Painted Drywall 61
Table A-1-3-1: Task 1 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Vinyl Flooring 62
Table A-1-3-2: Task 1 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Glass 63
Table A-1-3-3: Task 1 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Melamine 64
Table A-1-3-4: Task 1 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Painted Drywall 65
Table A-1-4-1: Task 1 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Vinyl Flooring 66
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Table A-1 -4-2:
Task 1 Results Dispatch® Hospital Cleaner Disinfectants Towels with Bleach/Glass
67
Table A-1-4-3:
Task 1 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Melamine
68
Table A-1-4-4:
Task 1 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Painted Drywall
69
Table A-2-1-1:
Task 2 Results Clorox Healthcare® Bleach Germicidal WipeA/inyl Flooring
70
Table A-2-1-2:
Task 2 Results Clorox Healthcare® Bleach Germicidal Wipe/Glass
70
Table A-2-1-3:
Task 2 Results Clorox Healthcare® Bleach Germicidal Wipe/Melamine
71
Table A-2-1-4:
Task 2 Results Clorox Healthcare® Bleach Germicidal Wipe/Painted Drywall
71
Table A-2-2-1
Task 2 Results Hype-Wipe® Bleach ToweletteA/inyl Flooring
72
Table A-2-2-2:
Task 2 Results Hype-Wipe® Bleach Towelette/Glass
72
Table A-2-2-3:
Task 2 Results Hype-Wipe® Bleach Towelette/Melamine
73
Table A-2-2-4:
Task 2 Results Hype-Wipe® Bleach Towelette/Painted Drywall
73
Table A-2-3-1
Task 2 Results Sani-Cloth® Bleach Germicidal Disposable WipeA/inyl Flooring
74
Table A-2-3-2:
Task 2 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Glass
74
Table A-2-3-3:
Task 2 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Melamine
75
Table A-2-3-4:
Task 2 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Painted Drywall
75
Table A-2-4-1
Task 2 Results Dispatch® Hospital Cleaner Disinfectants Towels with Bleach/Vinyl Flooring
76
Table A-2-4-2:
Task 2 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Glass
76
Table A-2-4-3:
Task 2 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Melamine
77
Table A-2-4-4:
Task 2 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Painted Drywall
77
Table A-3-1-1:
Task 3 Results Clorox Healthcare® Bleach Germicidal Wipe/Vinyl Flooring
78
Table A-3-1-2:
Task 3 Results Clorox Healthcare® Bleach Germicidal Wipe/Glass
78
Table A-3-1-3:
Task 3 Results Clorox Healthcare® Bleach Germicidal Wipe/Melamine
79
Table A-3-1-4:
Task 3 Results Clorox Healthcare® Bleach Germicidal Wipe/Painted Drywall
79
Table A-3-2-1
Task 3 Results Hype-Wipe® Bleach Towelette/Vinyl Flooring
80
Table A-3-2-2:
Task 3 Results Hype-Wipe® Bleach Towelette/Glass
80
Table A-3-2-3:
Task 3 Results Hype-Wipe® Bleach Towelette/Melamine
81
Table A-3-2-4:
Task 3 Results Hype-Wipe® Bleach Towelette/Painted Drywall
81
Table A-3-3-1
Task 3 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Vinyl Flooring
82
Table A-3-3-2:
Task 3 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Glass
82
Table A-3-3-3:
Task 3 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Melamine
83
Table A-3-3-4:
Task 3 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Painted Drywall
83
Table A-3-4-1
Task 3 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Vinyl Flooring
84
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Table A-3-4-2: Task 3 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Glass 84
Table A-3-4-3: Task 3 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Melamine 85
Table A-3-4-4: Task 3 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Painted Drywall 85
Table B-1: Decontamination Test Results 86
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List of Figures
Figure 2-1. Easel Used to Hold Coupons 7
Figure 3-1. Aerosol Deposition Apparatus 12
Figure 3-2. MDI and Actuator 12
Figure 3-3. "Hot Spot" Sections Inoculated (shown highlighted) on 42" x 42" Coupons 13
Figure 3-4. Clorox Healthcare® Bleach Germicidal Wipe Canister 14
Figure 3-5. Folding ofWipe for Wipe Sampling Step 1 (horizontal wiping pathway) 16
Figure 3-6. Horizontal Wiping Pathway 16
Figure 3-7. Folding ofWipe for Wipe Sampling Step 2 (vertical wipe sampling pathway) 17
Figure 3-8. Vertical Wiping Pathway 17
Figure 3-9. Folding ofWipe for Wipe Sampling Step 3 (diagonal wipe sampling pathway) 17
Figure 3-10. Diagonal Wiping Pathway 18
Figure 3-11. Folding ofWipe for Wipe Sampling Step 4 (perimeter wipe sampling pathway) 18
Figure 3-12. Perimeter Wiping Pathway 18
Figure 4-1. Residual Bleach Recovered from Melamine and Glass Material 26
Figure 4-2. Residual Bleach Recovered from Vinyl Flooring and Painted Drywall Material 27
Figure 4-3. Percent Liquid Dispensed on Material as a Function of Material Type, for Hype-Wipe® 27
Figure 4-4. Temporal Residual Bleach Recovered following a Decontamination Event using Clorox® Bleach
Wipe and Hype-Wipe® Bleach Towelette 29
Figure 4-5. Temporal Residual Bleach Recovered Following a Decontamination Event using Sani-Cloth®
Wipe and Dispatch® Towel with Bleach 29
Figure 4-6. Effect of Application Pressure on Liquid Dispensed Using Clorox® Bleach Wipe and Hype-
Wipe® Bleach Towelette 30
Figure 4-7. Effect of Application Pressure on Liquid Dispensed using Sani-Cloth® Wipe and Dispatch®
Towel with Bleach 31
Figure 4-8. Spatial Post-Decontamination Residual Spore Concentration Using Hype-Wipe® Bleach
Towelette /Glass Material at Different Inoculation Locations. 35
Figure 4-9. Spatial Post-Decontamination Residual Spore Concentration using Clorox Healthcare®
Wipe/Glass Material at Different Inoculation Locations 36
Figure 4-10. Spatial Post-Decontamination Residual Spore Concentration using Hype-Wipe® Bleach
Towelette /Painted Drywall Material at Different Inoculation Locations 37
Figure 4-11. Spatial Post-Decontamination Residual Spore Concentration using Clorox Healthcare®
Wipe/Painted Drywall Material at Different Inoculation Locations 38
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Acronyms and Abbreviations
ADA
aerosol deposition apparatus
ANOVA
Analysis of Variance
ASTM
American Society for Testing and Materials, now ASTM International
ATCC
American Type Culture Collection
B.
Bacillus
Bg
Bacillus globigii
°C
degrees Celsius
Cdiff
Clostridium difficile
CFU
colony forming unit(s)
CI
confidence interval
cm
centimeter(s)
cm2
square centimeter
CMAD
Consequence Management Advisory Division
CoC
chain of custody
Dl
deionized
DPG
(U.S. Army) Dugway Proving Ground
DQI
data quality indicator
DQO
data quality objective
DTRL
Decontamination Technologies Research Laboratory
EPA
U. S. Environmental Protection Agency
ft
foot (feet)
ft2
square foot (feet)
h
hour
H2O2
hydrogen peroxide
ID
identification
in
inch(es)
ISO
International Organization for Standardization
LR
log reduction
m2
square meter(s)
MDI
metered dose inhaler
mg
milligram(s)
Min
minute(s)
MOP
miscellaneous operating procedure
NHSRC
National Homeland Security Research Center
NIST
National Institute of Standards and Technology
OEM
Office of Emergency Management
OPP
Office of Pesticide Programs
ORD
Office of Research and Development
OSB
oriented strand board
pAB
pH-adjusted bleach
PARTNER
Program to Align Research and Technology with the Needs of Environmental Response
PBST
phosphate-buffered saline with Tween 20
PI
principal investigator
PPm
part(s) per million
PRB
polyester rayon blend
QA
Quality Assurance
QAPP
Quality Assurance Project Plan
QC
Quality Control
RH
relative humidity
StDev
standard deviation
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STS sodium thiosulfate
TSA tryptic soy agar
VHP vaporous hydrogen peroxide
WACOR work assignment contract officer representative
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Executive Summary
The likelihood of a biological incident such as the 2001 U.S. Postal Service system facilities contamination
through the introduction of a few letters containing Bacillus anthracis spores prompted the U.S.
Environmental Protection Agency (EPA), in a coordinated effort with other federal agencies, to develop
strategies, guidelines, and plans for decontamination of facilities and equipment to mitigate the risks of
contamination following a biological weapons attack.
As a part of the 2001 remediation efforts, various surface cleaning methods were used in buildings, including
those buildings that were fumigated prior to the clearance for reoccupation and normal use. These "low-tech"
methods included combinations of disposal of contaminated items, vacuuming, and the use of liquid
sporicides such as a pH-adjusted bleach (pAB) solution.
The use of commercially-available sporicidal wipes was presented as an effective decontamination technique
for the complete removal/inactivation of Bacillus (B.) atrophaeus spores [1 ], a surrogate for Bacillus
anthracis from a 12 inch (in) by 12 in material. This report specifically addresses the results of the
operational aspects of decontamination by sporicidal wipes and evaluates the efficacy of a single wipe for
the removal/inactivation of B. atrophaeus spores under varied conditions of application (type of sporicidal
wipe, material type, and application pressure) for larger surface areas tested (one to nine square feet (ft2))
under conditions that mimic their potential realistic field use.
The operational aspects of the wiping decontamination technique were evaluated, using four commercially
available sporicidal wipes, on painted drywall, vinyl tile, melamine board, and glass. All four sporicidal wipes
contained sodium hypochlorite ("bleach") as the active ingredient. The use of individually packaged, single-
use Hype-Wipe® Bleach Towelettes resulted in a higher volume of bleach dispensed, for all types of
materials tested, compared to the other wipes that are sold in bulk canisters. The bulk canister wipes are also
more susceptible to moisture loss because the canister lid must be opened to access individual wipes. The
amount of liquid dispensed increased with the size of the surface area that was wiped. However, the amount
of liquid bleach dispensed per unit area decreased when larger areas were wiped. Among the four materials
tested, the least amount of liquid bleach was transferred to glass, while painted drywall had the propensity to
retain the most liquid.
Another prerequisite for the high inactivation of spores would be the need to keep the surface wet for at least
five minutes [1], The residual amount of liquid remaining on the surface after a five-minute contact time was
measured and compared to the amount of liquid present on the surface without the five-minute contact time.
All surfaces were considered "wet" after the five-minute contact time, based on mass recovered after five
minutes. Liquid solutions dispensed from Hype-Wipe® Bleach Towelettes, and to a lesser extent from Clorox
Healthcare® Bleach Germicidal Wipes, were found to be least susceptible to losses due to evaporation or
absorption into the material.
A higher pressure applied during wiping resulted in higher liquid solution volume dispensed compared to
slight (low) pressure for all material/wipe type combinations. For these particular tests, the effect of applying
higher pressure (compared to slight pressure) resulted in an increase of approximately 60% in liquid being
dispensed from the Hype-Wipe® Bleach Towelettes to the coupon and up to a 300% increase in liquid from
the Sani-Cloth® wipes.
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The decontamination efficacy of a subset of two commercially available bleach wipes (Hype-Wipe® Bleach
Towelette, and Clorox Healthcare® Wipe) was evaluated on a medium size surface area of 12.25 ft2 (1.36
square meters (m2)) for different inoculation methods (hot spot versus broad area) and wipe application
pressure method (slight versus high). Such a 3.5 foot (ft) * 3.5 ft surface area is considered to be
representative of an area that can be wiped by a single person without physically relocating to a different
position.
The results indicate that the Hype-Wipe® Bleach Towelette was more effective in inactivating Bg spores than
the Clorox Healthcare® Wipe, both from glass and drywall surfaces. For glass, the overall log reductions in
spores by decontamination with the Hype-Wipe® Bleach Towelette and Clorox Healthcare® Wipe were 4.3 ±
0.6 and 3.4 ± 1.0, respectively, while for painted drywall, the log reductions were lower, namely, 3.6 ± 0.9 and
2.5 ±0.9, respectively. The Hype-Wipe® Bleach Towelette may have been more effective as it dispenses
more sporicidal liquid per decontamination wipe than the Clorox Healthcare® Wipe. The wipe application
pressure seems to have little or no effect on the decontamination efficacy of the wipe, despite more liquid
being dispensed from either decontamination wipe. The higher applied pressure may only have caused a
more pronounced redistribution of spores without improvement in overall efficacy of wiping the 12.25
square foot (ft2) surface area.
Three of the twenty four tests came back with a significant number (>500) of viable spores on the
procedural blank or negative control. Such contamination of blanks negates the ability to demonstrate a 6
log kill in viable spores. However, other tests clearly show the inability of either sporicidal wipe to inactivate
all spores from a 3.5 ft * 3.5 ft surface area. As such this contamination does not impact the overall
conclusions of this study.
The decontamination wipes that were used during the decontamination process were stored for a minimum
of 24 hours (h) at 4 degrees Celsius (°C) in sterile specimen cups. No viable Bacillus globigii (Bg) spores
were detected on these used wipes after the 24 h wait period. This implies that collected spores on the wipes
were inactivated
Spatial distributions of the post-decontamination spore concentration on the target coupons, as determined
from discrete samples, showed that cross contamination occurs during the decontamination wiping. The
likelihood of cross contamination increases as the towelette moisture content decreases. The folding of the
towelette when switching wiping motions exposes a fresh sporicidal solution on the wipe. However, the
same decrease in moisture content results in further redistribution of spores over areas that are already
cross contaminated. The decontamination scheme approach used in this study, as optimized for sampling
for a 1 ft2 (929 square centimeter (cm2)) area [1], was found to be unsuitable for larger 12.25 ft2 (1.36 m2)
surface areas. Further laboratory testing should be conducted in which sporicidal wipes are used only as
long as the exposed surface remains wet to reduce cross contamination of material and wipe surfaces.
Impact of the Study:
In contrast to the findings of a recent study that evaluated sporicidal wipes on 1 ft * 1 ft surface areas [1],
the current study demonstrates that sporicidal wipes were unable to achieve a 6 log reduction in viable B.
atrophaeus spores on material surfaces of 3.5 ft x 3.5 ft size when a single sporicidal wipe was used. A
reduction in wiped area per sporicidal wipe or use of multiple wipes is expected to improve the overall
efficacy. However, such an approach was not evaluated as part of this study.
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1 Introduction
1.1 Background
This project supports the mission of the U.S. Environmental Protection Agency (EPA) Office of Research and
Development's (ORD) Homeland Security Research Program (HSRP) by providing information relevant to
the decontamination of equipment or areas contaminated as a result of biological contamination incident.
Homeland Security Presidential Directive (HSPD)-10 tasked the U.S. Department of Homeland Security
(DHS) with coordinating the appropriate federal departments and agencies to develop comprehensive plans
that "provide for seamless, coordinated federal, state, local, and international responses to a biological
attack." As part of these plans, EPA, in conjunction with DHS and other agencies, is "developing strategies,
guidelines, and plans for decontamination of persons, equipment, and facilities" to mitigate the risks of
contamination following a biological weapons attack. EPA's National Homeland Security Research Center
(NHSRC) provides expertise and products that can be widely used to prevent, prepare for, and recover from
public health and environmental emergencies arising from terrorist threats and incidents.
In 2001, the introduction of letters containing Bacillus anthracis (causative agent of anthrax) spores into the
U.S. Postal Service system resulted in the contamination of several facilities. Although most of the facilities in
which these letters were processed or received in 2001 were heavily contaminated [2], the facilities were
successfully remediated with approaches such as fumigation with chlorine dioxide or vaporous hydrogen
peroxide (VHP®). It is well agreed that available, effective and economical decontamination methods having
the capacity to be employed over wide areas (outdoor and indoor) are required to increase preparedness for
such a release.
Prior to the fumigation used in heavily contaminated facilities, surface cleaning methods were used in
primarily contaminated facilities and secondarily contaminated areas (e.g., cross contaminated letters that
came in contact with the anthrax spores contained in letters or tracked from primarily contaminated sites) or
showing a minimal presence of the anthrax spores. These "low-tech" methods included combinations of
disposal of contaminated items, vacuuming, and the use of liquid sporicides such as a pH-adjusted bleach
(pAB) solution.
1.2 Project Objectives and Process
The primary objective of this project was to provide agencies responding to occurrences of biological
contamination with operational criteria that would lead to efficacious decontamination of various
materials/surfaces contaminated with Bacillus anthracis. This report specifically addresses the results of
investigating four commercially available sporicidal wipes for potential use in the removal/inactivation of B.
anthracis spores under varied conditions of application (type of sporicidal wipe, type of surface, application
pressure, and size of area). This study was conducted in two phases: the first phase addressed the
operational aspects of decontamination using sporicidal wipes by measuring the wetting ability of four
different sporicidal wipe products whose efficacies had previously been evaluated by Meyer et al. [1], The
second phase of the study consisted of evaluating two of the four sporicidal wipes for their decontamination
efficacy of medium-size contaminated surfaces, following inoculation of various material surfaces with B.
atrophaeus spores, which served as a surrogate for B. anthracis.
1
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2 Experimental Approach
This section describes the project, the testing program, test materials, test facilities and equipment, general
decontamination approach and test conditions, and the methods that were used to evaluate the data related
to the project objectives. Testing was conducted in a spray chamber located in High Bay Room H122A at
EPA's facility in Research Triangle Park, North Carolina.
2.1 Project Description
2.1.1 PHASE 1: Operational Aspects of Decontamination by Sporicidai Wipes
A preliminary study was conducted to develop the operational criteria that would be most effective for
decontamination of medium size (1-9 ft2) surfaces by wiping the surfaces with sporicidai (decontamination)
wipes. Four different commercially-available decontamination wipes were evaluated for their ability to wet
the surfaces of four common porous/nonporous materials (painted drywall, vinyl tile, melamine board, and
glass) of various sizes by measuring the amount of decontamination liquid remaining on the surface
following wipe application.
The amount of liquid remaining on each material coupon following wipe application was determined
gravimetrically, using the weight loss of the wipe. The effects of wiping pressure on the weight loss of the
wipe was also evaluated. Additionally, the gravimetric loss of liquid from the wiped coupon surface was
determined over a five-minute period. This mass of liquid remaining on a coupon after five minutes (min)
could be considered an indicator of the relative efficacies of the decontamination wipes since inactivation
of bacterial spores is not expected to occur on a dried surface. Results of this preliminary study aided in
the selection of two sporicidai wipes for use in Phase 2 of the project. Phase 2 was designed to evaluate
the effectiveness of sporicidai wipes in decontaminating surfaces inoculated with bacterial spores.
The four commercially available sporicidal/antimicrobial wipes examined in this preliminary study were:
1. Clorox Healthcare® Bleach Germicidal Wipe
2. Sani-Cloth® Bleach Germicidal Disposable Wipe
3. Dispatch® Hospital Cleaner Disinfectant Towel with Bleach
4. Hype-Wipe® Bleach Towelette.
Relevant physical and chemical properties of these wipes are summarized in Table 2-1.
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Table 2-1. Sporicidal Wipe Information
Product
Vendor
Size
Active
Ingredients
Components
EPA
Registrationa
Minimum
Contact
Time (min)
Clorox Healthcare®
Bleach Germicidal
Wipe
Clorox® Professional
Products Co.
6.75 in x 9 in
(17.1 cm x 22.8 cm)
Sodium
hypochlorite
Sodium hypochlorite 0.1-1.0%,
sodium metasilicate 0.1-1.0%,
sodium hydroxide 0.1-1.0%
67619-12"
3C
Sani-Cloth® Bleach
Germicidal Disposable
Wipe
Professional
Disposables
International, Inc.
6 in x 10.5 in
(15.2 cm x 26.7 cm)
Sodium
hypochlorite
Trisodium phosphate
dodecahydrate 1-2.5%,
sodium hypochlorite <1 %
9480-8"
4C
Dispatch® Hospital
Cleaner Disinfectants
Towels with Bleach
Clorox® Professional
Products Co.
6.75 in x 8 in
(17.1 cm x 20.3 cm)
Sodium
hypochlorite
Sodium hydroxide <0.2%, sodium
metasilicate <0.6%, sodium
hypochlorite <1.0%
56392-8"
5C
Hype-Wipe® Bleach
Towelette
Current
Technologies Inc.
6 in x 12 in
(15.2 cm x 30.4 cm)
Sodium
hypochlorite
Single-use Towelettes saturated
with 0.525% sodium hypochlorite
(a 1:10 dilution)
70590-1"
4C
a Registration with EPA's Office of Pesticide Programs (OPP) indicates EPA/OPP has evaluated the data provided by the
manufacturer to show the product is effective and has no unreasonable adverse effects on humans, the environment, and non-
target species and has issued a registration or license for use in the United States,
k Wipe has been registered as sporicidal against Clostridium difficile but not specifically for use against B. anthracis.
0 Contact time reflects the prescribed time a surface has to be wet to kill Clostridium difficile (C. diffi).
2.1.2 PHASE 2: Evaluation of Efficacy of Hypochlorite Wipes
The objective of this task was to evaluate the efficacy of two commercially available hypochlorite wipes to
decontaminate material coupons with medium-size surface areas (i.e., 42 in x42 in = 12.25 ft2) contaminated
with B. atrophaeus spores (1 * 107 colony forming units (CFU)). Work conducted in the Phase I preliminary
study measured the wetting ability of four different sporicidal wipe products with efficacies previously
evaluated by Meyer et al.[1]. Two of the four hypochlorite wipes of different wetting abilities were evaluated
for their decontamination efficacy on two different indoor materials, namely, glass and painted drywall. Glass
represents nonporous indoor materials, while painted drywall represents a more porous indoor material. The
two hypochlorite wipes selected for the decontamination efficacy experiments were:
a. Clorox Healthcare® Bleach Germicidal Wipe
b. Hype-Wipe® Bleach Towelette.
The relevant physical and chemical properties of each of these two sporicidal wipes are summarized in Table
2-1.
2.2 Testing Program Description
2.2.1 PHASE 1: Operational Aspects of Decontamination by Sporicidal Wipes
The operational aspects of the wiping decontamination technique were evaluated through three different
tasks. Task 1 consisted of determining the amount of liquid remaining on each material coupon of known
3
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surface area following wipe application. Task 2 evaluated the loss of liquid from coupon surfaces over a
required five-minute contact time period following wipe application. Task 3 evaluated the influence on the
results in relation to the applied pressure when wiping the surface. These three tasks, described in the
following sections, were used as indicators of the relative efficacies of the decontamination wipes.
2.2.1.1 Task 1 - Determination of Effective Surface Coverage ofSporicidai Wipes
Four different decontamination wipes were evaluated to determine how much surface area could potentially
be wiped with a single wipe, based on whether the surface remained wet for required time. A single wipe was
used to wipe surfaces ranging in size from 1.0 to 9.0 ft2 of each material type. Each surface was wiped and
each wipe was refolded according to the patterns and directions described in Section 3.6.3. To ensure
consistency in wipe pattern and wiping pressure applied, the same person performed this operation across
all Task 1 tests. The amount of liquid dispensed by each type of wipe on a target surface area for each
specific type of material was determined by measuring its weight loss following the decontamination wiping
of the surface. Table 2-2 provides the Task 1 test matrix that was repeated for all four decontamination
wipe types. Five replicate samples were tested for each type of wipe/material/surface area combination.
The largest area wiped under Task 1 was limited to 3 ft x 3 ft. This was a smaller area than the actual
decontamination testing area of 3.5 ft x 3.5 ft (Phase 2) that was established after completion of Phase 1
testing.
Table 2-2. Task 1 Test Matrix Conducted for Each Type ofSporicidai Wipe
Test ID
Material
Coupon Area Sampled per Wipe
(ft x ft)
Replicates
1x1
1.5x1.5
1-D
Painted Drywall
2x2
5 per coupon area
2.5x2.5
3x3
1x1
1.5x1.5
1-V
Vinyl Tile
2x2
5 per coupon area
2.5x2.5
3x3
1x1
1.5x1.5
1-M
Melamine Board
2x2
5 per coupon area
2.5x2.5
3x3
1-G
Glass
1x1
5 per coupon area
1.5x1.5
4
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Test ID
Material
Coupon Area Sampled per Wipe
(ft x ft)
Replicates
2x2
2.5x2.5
3x3
2.2.1.2 Task 2 - Surface Retention Times of Wipe Decontaminant Liquid
Decontamination wipes have a manufacturer-recommended contact time for C. diff. This task was designed
to determine how much decontamination liquid remained on a horizontal coupon surface for a set time period
following the wiping application. Coupons of all four materials were wiped with the decontamination wipes
and then allowed to dry for five minutes. Determination of the amount of liquid remaining on a coupon after
the five-minute period was accomplished by sampling the surface with one or more dry (non-sporicidal) wipes
and measuring the liquid recovery based on weight gain of the dry wipe(s). The amount of decontamination
liquid lost from the coupon during the five-minute drying period was assessed by comparing the weight gain
of these dry wipes after the five-minute contact time to the weight gain of the same dry wipes used to sample
the coupons immediately after the decontamination wipe procedure. One ft2 coupons of each material were
used in this task. Table 2-3 shows the Task 2 test matrix that was repeated for all four decontamination wipe
types.
Table 2-3. Task 2 Test Matrix per Sporicidal Wipe Type
Test ID
Material
Drying Time
(minutes)
Replicates
2-D
Painted Drywall
0
5
5
2-V
Vinyl Tile
0
5
5
2-M
Melamine Board
0
5
5
2-G
Glass
0
5
5
2.2.1.3 Task 3 - Impact of Wiping Pressure
The amount of decontamination solution dispensed by each type of wipe on a coupon surface, as a function
of the pressure applied by the sampling person during the wiping process, was examined in this task. The
four types of decontamination wipes were evaluated on the four different material surfaces to determine if
application pressure affected the amount of decontamination solution dispensed onto a coupon. A single
decontamination wipe was used per coupon and was applied using slight or higher pressure by the same
sampling personnel. The change in weight of the decontamination wipe was measured immediately after
wiping. The forces applied were not quantified, yet are referred to as "slight" and "heavy" to indicate relative
5
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pressure. One ft2 coupons of each material were used for this task. Table 2-4 shows the Task 3 test matrix
that was repeated for all four decontamination wipe types.
Table 2-4. Task 3 Test Matrix per Sporicidal Wipe Type
Test ID
Material
Pressure Applied
to Wipe
Replicates
3-D
Painted Drywall
- Slight pressure
- Heavy Pressure
5
3-V
Vinyl Tile
- Slight pressure
- Heavy Pressure
5
3-M
Melamine Board
- Slight pressure
- Heavy Pressure
5
3-G
Glass
- Slight pressure
- Heavy Pressure
5
2.2.2 PHASE 2: Evaluation of Efficacy of Hypochlorite Wipes
The testing approach for evaluation of decontamination methods involved wiping of contaminated surfaces
following inoculation of various material surfaces with B. atrophaeus spores, which served as a non-
pathogenic surrogate for B. anthracis. B. atrophaeus spores are also known to be valid surrogates for
decontamination studies involving hypochlorite solutions [3], Decontamination wiping was performed using
hypochlorite-impregnated wipes. Individual sections of the medium size surface were sampled for residual
spores after a contact time of (at least) 30 min between the residual sporicidal liquid as dispensed from the
decontamination wipe and the vertical surface. Recovery of spores from coupon surfaces following the
decontamination technique was measured by plating the extracts from the wipes used for sampling. Aliquots
from the extract liquids were spread plated using serial dilutions onto culture media and following an 18 to 24
h incubation at 35 °C ± 2 °C, the media plates were enumerated and the CFU were quantified. Results of test
samples were compared to the results of positive control samples consisting of contaminated surfaces that
had not been subjected to decontamination.
Two commercially available hypochlorite wipes were evaluated to determine their decontamination efficacy
on the nine 14 in x 14 in inoculated sections of the 42 in x42 in glass and painted drywall coupon surfaces. A
single decontamination wipe was used to decontaminate the 42 in by 42 in surface using the same wiping
method established in the preliminary study (Section 2.2.1). While the test coupons were placed horizontally
for spore inoculation, the coupons were placed vertically for both the wipe decontamination procedure and
the subsequent wipe sampling of residual spores on the surfaces. Coupons were maintained in an upright
position using a heavy duty easel (Lyptus Wood "Dulce" Easel, Richeson Art, Kimberly, Wl), as shown in
Figure 2-1. The easel was modified slightly to better accommodate both the coupons and dry Polyester
Rayon Blend (PRB) wipes placed below the coupons. These wipes were added to capture potential runoff of
wipe solution from the near vertical surface. The easel was sterilized using Dispatch® wipes (Clorox®
Professional Products Co. Oakland, CA). Five minutes after the Dispatch sterilization, the easel was wiped
again with a separate wipe soaked with 3% sodium thiosulfate (STS) to neutralize/remove any remaining
6
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hypochlorite, and then again with an alcohol wipe (VWR® Pre-moistened Clean-Wipes™ with Isopropyl
Alcohoi/Deionized Water, VWR, Radnor PA).
Figure 2-1. Easel Used to Hold Coupons
Variables included in this decontamination test matrix and the corresponding test codes are listed below.
1. Decontamination Wipe
a. Clorox Healthcare® (C)
b. Hype-Wipe® (H)
2. Inoculation Method
a. Hot Spot upper left corner(inoculation location a) (1)
b. Hot Spot center (inoculation location e) (2)
c. Hot Spot lower right corner (inoculation location i) (3)
d. Broad area, all nine sections (0)
3. Material
a. Glass (G)
b. Painted Drywall (D)
4. Application Variations
a. Slight Pressure application (1)
b. Higher Pressure application (2)
Assigned test identifications (IDs) were contaminated in the order shown in Table 2-5. For example,
decontamination test C2D1 indicates the use of a Clorox Healthcare® wipe (C) to wipe the 42"x42" surface
that was inoculated in the center (2). The material was painted drywall (D) and slight pressure (1) was
applied during the wiping of the surface. The target inoculation for the Hot Spot testing was 1 x 107 spores
per ft2, while the target inoculation for the Broad Area testing was 1 x 106 spores per ft2. Note that only the
7
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center 12 in x 12 in of a 14 in x 14 in coupon is inoculated, as the surrounding one-inch border is covered
during the inoculation process.
For both Hot Spot and Broad Area decontamination wiping, the upper left section (section a) of the coupon
was designated as the first corner to receive the decontamination wipe, and the lower right section (i) was the
last section to receive the decontamination wipe (see Section 3.3). The complete decontamination matrix
involved 24 separate conditions, as depicted in Table 2-5 by test ID.
Table 2-5. Decontamination Test Matrix for Wiping Surfaces with Sporicidal Wipes
Test Code
Material
Inoculation
Method and
Location
Decontamination Wipe
Used
Pressure Applied
C1G1
Glass
a
Clorox Healthcare®
Slight
C1D1
Painted drywall
a
Clorox Healthcare®
Slight
C2D1
Painted drywall
e
Clorox Healthcare®
Slight
C2G1
Glass
e
Clorox Healthcare®
Slight
C3G1
Glass
i
Clorox Healthcare®
Slight
H0D1
Painted drywall
Broad Area
Hype-Wipe®
Slight
H0D2
Painted drywall
Broad Area
Hype-Wipe®
Heavy
H1D1
Painted drywall
a
Hype-Wipe®
Slight
H1G1
Glass
a
Hype-Wipe®
Slight
H2G1
Glass
e
Hype-Wipe®
Slight
H3D1
Painted drywall
i
Hype-Wipe®
Slight
H3G1
Glass
i
Hype-Wipe®
Slight
H3D2
Painted drywall
i
Hype-Wipe®
Heavy
H3G2
Glass
i
Hype-Wipe®
Heavy
C3G2
Glass
i
Clorox Healthcare®
Heavy
H2D1
Painted drywall
e
Hype-Wipe®
Slight
C3D1
Painted drywall
i
Clorox Healthcare®
Slight
C0G1
Glass
Broad Area
Clorox Healthcare®
Slight
C0G2
Glass
Broad Area
Clorox Healthcare®
Heavy
C3D2
Painted drywall
I
Clorox Healthcare®
Heavy
H0G1
Glass
Broad Area
Hype-Wipe®
Slight
H0G2
Glass
Broad Area
Hype-Wipe®
Heavy
C0D1
Painted drywall
Broad Area
Clorox Healthcare®
Slight
C0D2
Painted drywall
Broad Area
Clorox Healthcare®
Heavy
8
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2.3 Definitions of Effectiveness
The surface decontamination efficacy for each decontamination technique and surface material combination
was evaluated by measuring the difference in the logarithm of the measured CFU before decontamination
(determined from sampling of the positive control coupons) and after decontamination (determined from
sampling of the test coupons) for that material. This value is reported as a log reduction (LR) on the specific
material surface as defined in Equation 2-1.
iVc Nt
2 log (CFUck) 2 log (CFUsk)
n = — — (2-1)
'' Nc Ns
where:
Surface decontamination effectiveness; the average log
1j. = reduction (LR) of spores on a specific material surface (surface
material designated by /)
The average of the logarithm (or geometric mean) of the
^ 1 °S(CFUc k) ^ number of viable spores (determined by CFU) recovered on the
— control coupons (C indicates control and Nc is the number of
control coupons)
k=l
Nc
The average of the logarithm (or geometric mean) of the
V log(CFU k) number of viable spores (determined by CFU) remaining on the
k=1 = surface of a decontaminated coupon (S indicates a
Ns decontaminated coupon and Ns is the number of coupons
tested).
When no viable spores were detected, a value of 0.5 CFU was assigned for CFUs.k, and the efficacy was
reported as greater than or equal to the value calculated by Equation 2-1.
The standard deviation of the average LR of spores on a specific material (P|j) is calculated by Equation 2-2:
SDn--
N,
' (2-2)
Ns-1
where:
9
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Standard deviation of l"|i, the average LR of spores on a specific
^ material surface
n _ The average LR of spores on a specific material surface
'' (surface material designated by /)
The average of the LR from the surface of a decontaminated
Xk =
coupon (Equation 2-3)
Ns = Number of test coupons of a material surface type.
N,
2((log(CF[/J-log(CTI/„,.))
xk = — (2-3)
N,
Where:
Represents the "mean of the logs" (geometric mean), the
nc average of the logarithm-transformed number of viable
2>g(Cfl/cl)
spores (determined by CFU) recovered on the control
log(CFUc) = — — coupons (C = control coupons, Nc = number of control
coupons, k = test coupon number and Ns is the number
of test coupons)
Nc
CFUsk =
Ns
Number of CFU on the surface of the kth decontaminated
coupon
Total number (1,k) of decontaminated coupons of a
material type.
In this report, decontamination efficacy is generally reported in terms of LR for a particular material. Results
may include whether the average LR for a particular test is > 6.0, since a decontaminant that achieves > 6 LR
(against a 6-7 log challenge) is generally considered effective during laboratory efficacy tests. It is important
to note that demonstrated effectiveness in the laboratory can help predict the performance of a
decontamination procedure/product in the field, but the 6 LR benchmark should not be considered in direct
connection with achieving specific clearance goals.
10
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3 Material and Methods
3.1 Test Coupon Preparation and Sterilization
Four types of materials (painted dry wall, vinyl tile, melamine, and glass) were used for Phase 1 testing, while
only painted dry wall and glass were used for Phase 2. These four types of materials were selected as being
typical of materials commonly used in buildings and meeting industry standards or specifications for indoor
use in terms of quality, surface characteristics, and structural integrity. Uniformity among the test coupons of
a given material was achieved by obtaining and preparing a quantity of material sufficient to allow multiple
test coupons to be prepared with presumably uniform characteristics. Coupons were re-used for the various
tests after being subjected to a thorough and consistent drying and surface cleaning process following each
use. The coupons were cut to the required sizes and sterilized before use. Stainless steel contamination
control coupons (14 in x 14 in) were prepared by using heavy duty power hydraulic shears to cut the metal
from larger sheets. These stainless steel coupons were sterilized prior to use by steam autoclaving.
For Phase 1, all coupons were cleaned with an alcohol wipe and air-dried immediately prior to use to remove
any incidental dust and grime that may have accumulated throughout the repeated use of the same
materials. Coupons for wiping efficacy tests (Phase 2) were sterilized with 400 parts per million (ppm)
hydrogen peroxide (H2O2) vapor for four h using a STERIS VHP® ED1000 (STERIS Life Sciences, Mentor,
OH) generator. Prior to use, the coupons treated with VHP® were kept at room temperature for a minimum
of 2-3 days to force off-gassing of residual H2O2 from the coupons so that biocidal activity was prevented.
1. Painted Drywall. Drywall coupons were cut to size from a 4 ft x 8 ft SHEETROCK Brand 1/2 in-
thick Drywall Panel (National Gypsum Company, Charlotte, NC) and painted with two coats of
white latex paint (Behr Premium Plus Interior Flat White Latex Paint, Home Depot, Durham, NC).
Test ID code D.
2. Vinyl Tile. Coupons were cut to size from Armstrong 12 ft-wide River pattern Staggered Slate
Brown Multi Vinyl Sheet (Model G4820, Home Depot, Durham, NC) and were glued to oriented
strand board (OSB). Test ID code V.
3. Melamine Boards. Coupons were cut to size from a 4 ft x 8 ft sheet of % in Melamine (Model
461877, Home Depot, Durham, NC). Test ID code M.
4. Glass. Glass coupons (3/16 in-thick tempered glass, Durham Glass, Durham, NC) were
purchased pre-cutto the required sizes. Test ID code G.
3.2 Bacillus Spore Preparation
The test organism for this work was a powdered spore preparation consisting of a mixture of B. atrophaeus
(formerly known as Bg) spores (American Type Culture Collection (ATCC) 9372) and silicon dioxide particles.
The preparation was obtained from the U.S. Army Dugway Proving Ground (DPG) Life Science Division. The
preparation procedure is fully reported in Brown et al. [4] and can be summarized as follows: After 80-90
percent sporulation, the suspension is centrifuged to generate a preparation of approximately 20 percent
solids. The final product in the form of a powdered matrix containing approximately 1 x 1011 viable spores per
11
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gram is prepared by dry blending and jet milling the dried spores with fumed silica particles (Degussa,
Frankfurt am Main, Germany).
The powdered preparation as received was loaded into metered dose inhalers (MDIs) at the EPA test site
according to a proprietary protocol. Control checks for each MDI were included in each batch of coupons
contaminated with a single MDI.
3.3 Coupon Inoculation Procedures
Coupons (test and positive controls) were inoculated independently with spores of B, atrophaeus by being
placed into a separate dosing chamber, called an Aerosol Deposition Apparatus (ADA), designed to fit one
14 in x 14 in coupon of any thickness [5], The ADA consisted of a stainless steel hood sized to cover the area
of a square test coupon exactly. In the center at the top of the hood was an opening to which an MDI and an
MDI actuator were attached. Photographs of an ADA, an MDI, and an MDI actuator used in this project are
shown in Figures 3-1 and 3-2.
Figure 3-1. Aerosol Deposition Apparatus Figure 3-2. MDI and Actuator
Prior to the start of any experiment, all components needed for the aerosol deposition of spores onto material
coupon surfaces using the ADA deposition approach must be sterilized and stored in a sterile environment
until use. ADAs and coupons were sterilized with a 250-ppm, 4-h VHP sterilization cycle, while the MDI
actuator, with attached MDI adaptor, was wiped with pAB, then rinsed with deionized (Dl) water. Sterilization
was not necessary for binder clips, MDIs, vortex, or the aerosol trap.
The MDI was discharged a single time into the ADA. The MDIs are claimed to provide 200 discharges per
MDI. The number of discharges per MDI was tracked so that use did not exceed this value. Additionally, the
weight of each MDI was determined after completion of the contamination of each coupon. The
contamination control coupons (14 in x 14 in stainless steel coupons) were inoculated as the first, middle,
and last coupons within a single group of coupons inoculated by any one MDI within a single test. Puffed
spores were allowed to settle onto the coupon surfaces for a minimum period of 18 h.
12
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For both Hot Spot and Broad Area inoculations, B. atrophaeus spores were inoculated onto 14 in x 14 in
sections of 42 in x42 in coupons. Dosing was conducted according to established National Homeland
Security Research Center (NHSRC) laboratory aseptic procedures described previously [6, 7],
For the Hot Spot tests, inoculation was performed at a target concentration of 1 x 107 spores/ft2 in one of the
nine 14 in x 14 in sections: the upper left corner (a), the center (e), and the lower right corner (i), as shown in
Figure 3-3.
Figure 3-3. "Hot Spot" Sections Inoculated (shown highlighted) on 42" x 42" Coupons
For the Broad Area testing, all nine (14 in x 14 in) sections were inoculated at a target concentration of 1 x
106 spores/ft2 for each section.
3.4 Decontamination Procedure
The decontamination procedure for all four decontamination wipes is described below:
1. Three of the four wipes were supplied in canisters. The Clorox Healthcare® Bleach Germicidal Wipe
is shown in Figure 3-4. Each canister was gently rolled and/or inverted 3-4 times to distribute liquid in
advance of removing Towelettes. Sterile gloves were used to remove the first 2-3 wipes from the
container and these wipes were discarded based on American Society for Testing and Materials,
now ASTM International (ASTM) 2896-12. Gloves were changed as necessary to maintain sterility.
Hype-Wipe® Bleach Towelettes were individually packaged and did not require this step.
2. The same wiping procedure utilized for sampling the coupon surfaces (as described in Section 3.6.3)
was used for the decontamination procedure.
3. Each decontamination wipe was stored in a separate sterile conical tube for at least 24 h after wipe
testing. After 24 h, the decontamination wipe was neutralized by adding 35 mL of phosphate-
buffered saline with Tween 20 (PBST) with 2.5% STS to the conical tube.
The steps to neutralize these specific decontamination wipes are based on the study by Meyer et al. [1],
13
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HEALTHCARE-
Bleach Germicidal Wipe;
^Kr^ilUQX Mtl *-» " M I
r. £¦— S
JW m trttaiiM>'»«s=u °iJ
Figure 3-4. Clorox Healthcare® Bleach Germicidal Wipe Canister
3.5 PHASE 1 Measurement Protocol
3.5.1 Sampling Approach
The sampling approaches used for the Phase 1 operational aspect testing and Phase 2 wipe
decontamination testing were identical (detailed in Section 3.6.3). During Phase 1, coupons from the same
material were chosen randomly for sampling to avoid a systematic bias that could have been introduced due
to personnel fatigue or changes in room environment. Each of the 25 coupons per material (five replicates,
five sizes) was assigned a random unique integer between 1 and 25. Wiping occurred in numerical order. A
Microsoft Excel 2013 random function coupled with the index and rank function was used to create a random
coupon number generator.
3.5.2 Gravimetric Measurements
The amount of liquid retained on the coupons following the decontamination wiping procedure was
determined by measuring the weights of the wipes before and after use. The following gravimetric
procedure for Tasks 1 and 3 was used:
1. Using a recently calibrated analytical balance, accurate to ±1 milligram (mg), tare the balance
using a new weigh boat and measure the weight of the wipe to the nearest 1 mg immediately
following removal from the bulk canister or individual package (Hype-Wipe® Bleach Towelettes
only).
2. Wearing gloves, use the wipe for decontamination of the coupon according to the procedure
detailed in Section 3.6.3.
3. Immediately place the used wipe in the same weigh boat used in Step 1 and determine the weight
of the used wipe to the nearest 1 mg.
4. The difference between weights in Step 1 and Step 3 is reported as the weight loss of the wipe
which is equated to the amount of liquid transferred to the coupon.
For Task 2, the following procedure was used.
14
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1. Tare the balance using a new weigh boat, and measure the combined weight of two dry wipes to
the nearest 1 mg immediately following removal from the bulk container (canister).
2. Remove one wipe from the boat and record the weight of the second wipe alone.
3. Use a two-wipe variation of the procedure outlined in Section 3.6.3 to sample the entire surface.
Start with the first wipe, using horizontal and vertical strokes, flipping the wipe over between
horizontal and vertical strokes.
4. Remove the second wipe from the weigh boat, place first wipe in the weigh boat, and record the
weight of the first wipe alone after use.
5. Use the second wipe to sample the entire surface with diagonal and perimeter strokes.
6. Immediately place the second wipe in the same weigh boat and determine the weight of both used
wipes together.
7. The difference between weights in Step 6 and Step 1 is reported as the weight gain and is equated
to the amount of decontamination liquid remaining on the coupon following the drying period.
Steps 2 and 4 were included initially to determine individual wipe weight increases as needed.
3.5.3 Method Verification
A procedural blank was obtained as a means of verifying the methods used for Tasks 1 and 3. A procedural
blank was acquired by handling a decontamination wipe as per the procedure but without applying the wipe
to a surface. The weight loss of the wipe determined in this manner was taken to be indicative of the amount
of decontamination liquid that remains on the glove.
The dry wipe sampling method was verified by applying 10 mL of a 5000 ppm hypochlorite solution in 1 mL
drops to the surface of the nonporous coupon materials and using the dry wipe method to measure recovery.
For verification, the recovery had to be within a range of 75% to 125% of the amount applied, with an
expected variance not exceeding 30%.
3.6 PHASE 2 Measurement Protocol
3.6.1 Sterile Handling of Wipes
New containers of decontaminant wipes were opened for each test day. The first three wipes were not
considered to be representative of the package and were discarded based on ASTM 2896-12 for small scale
Petri dish experiments, which stated, in part: "For multi-count containers only, use sterile gloves to remove 2-
3 wipes from the container and discard. For canisters, gently roll and/or invert 3-4 times to distribute liquid in
advance of removing towelettes. If possible, have another person hold the towelette container to avoid
contamination of the gloves when removing the wipes. Change gloves as necessary to maintain sterility."
3.6.2 Sampling Site Environmental Conditions
Ambient environmental conditions such as temperature, relative humidity (RH), and barometric pressure can
affect the evaporation rate of liquids from surfaces. Taking these influences into account was especially
important for Phase 1 testing. All tests were conducted at room temperature, ambient RH and ambient
barometric pressure. RH and temperature were monitored during testing, and tests were conducted on days
with environmental conditions within 35-60% RH and 20-25 °C. All coupons were conditioned under ambient
conditions for one week before use.
15
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3.6,3 Wipe Sampling Procedures
Within a single test, surface sampling of the materials was completed for all procedural blank coupons before
sampling of any test material was performed. Surface sampling was done by wipe sampling in accordance
with the protocols documented below. Prior to the sampling event, all materials needed for sampling were
prepared using aseptic techniques. The materials specific to the wipe sampling protocol are included in the
relevant sections below. The general sampling supplies were sterile or sterilized/disinfected for each
sampling event.
a, Don a fresh pair of latex or nitrile gloves for the wipe sampling procedure .
b. With gloved hand, grasp the wetted decontamination wipe. Using the other hand, gently fold the
wipe (Figure 3-5). Do not squeeze the wipe to avoid losses of the wetting solvent.
Figure 3-5. Folding of Wipe for Wipe Sampling Step 1 (horizontal wiping pathway)
c. Starting in the top left corner, wipe the surface horizontally, working downward, to cover the
surface completely. The horizontal wipe sampling pathway is shown in Figure 3-6.
Figure 3-6. Horizontal Wiping Pathway
—1
l*
f
'1
I
—1
1*
f
'1
*
—-1
r
**
*1
16
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d. Using both hands, gently refold the wipe so that that surface used for the horizontal wipe
sampling is now on the inside (Figure 3-7).
Figure 3-7. Folding of Wipe for Wipe Sampling Step 2 (vertical wipe sampling pathway)
e. Starting in the bottom left corner, wipe the surface vertically, working toward the right, to
cover the surface completely. The vertical wipe sampling pathway is shown in Figure 3-8.
Figure 3-8. Vertical Wiping Pathway
f. Using both hands, gently refold the wipe diagonally, so that that surface used for the
vertical wipe sampling is now on the inside (Figure 3-9).
Figure 3-9. Folding of Wipe for Wipe Sampling Step 3 (diagonal wipe sampling pathway)
17
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g. Starting in the top left corner, wipe the surface diagonally, working toward the bottom
right corner, to cover the surface completely. The diagonal wipe sampling pathway is
shown in Figure 3-10.
Figure 3-10. Diagonal Wiping Pathway
h. Using both hands, gentiy refold the wipe so that that surface used for the diagonal wipe
sampling is now on the inside (Figure 3-11).
Figure 3-11. Folding of Wipe for Wipe Sampling Step 4 (perimeter wipe sampling pathway)
i. Starting in any corner, wipe the perimeter of the coupon. The perimeter wipe sampling
pathway is shown in Figure 3-12.
Figure 3-12. Perimeter Wiping Pathway
18
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Following decontamination of the spore-inoculated coupons, each of the nine 14 in x 14 in sections (including
both inoculated and non-inoculated sections in the Hot Spot tests) of the 42 in x 42 in test coupon and the
nine 14 in x 14 in sections in a positive control coupon were sampled individually using PRB wipes in
accordance with the wipe sampling procedure described above. Sampling was performed after a coupon was
visually dry and no sooner than 30 minutes after decontamination wiping was complete.
Because the coupons were placed vertically on the easel, additional dry PRB wipes were placed under the
bottom edge of the coupon to capture any drips that might migrate downward and off the coupon during
wiping. These dry wipes were also considered to be samples for determination of viable spores.
Decontamination wipe samples were placed in 35 mL of PBST with 3% STS immediately following use.
LR was determined using the sum of viable spores collected on individual wipes from all sections of each
coupon and the wipes below the coupons as compared to the sum of viable spores collected on individual
wipes from positive control coupons not subjected to decontamination.
This standard wiping method was used to ensure high reproducibility of laboratory results as it ensured that
the whole surface area was wiped in a consistent manner. This test method may not reflect the performance
when used by responders. However, this approach reflects the best possible case when utilizing these
sporicidal wipes.
3.6.4 Swab Sampling
The general approach for swab sampling was to use a moistened swab (BactiSwab® Collection and
Transport System, Remel, Thermo Fisher Scientific, Waltham, MA) to wipe a specified area to recover
bacterial spores. Swab samples were collected from all decontamination procedure equipment before use to
serve as sterility checks.
3.6.5 Phase 1 Sample Identification
Each sample was assigned an ID based on the sample coding outlined in Table 3-1. This ID was used in
written records in laboratory notebooks documenting the surface area, surface type, pressure applied, etc.
19
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Table 3-1. Sample Coding for Phase 1
Coupon Identification: WA73-T-M-D-SS-N
Category
Example Code
Description
T
(Task)
(1,2,3)
Task under which the sample was collected
M
(Material)
V
G
M
D
Vinyl Tile
Glass
Melamine Board
Painted Drywall
D
(Decontamination Wipe)
1
2
3
4
Clorox Healthcare® Bleach Germicidal Wipe
Sani-Cloth® Bleach Germicidal Disposable Wipe
Dispatch® Hospital Cleaner Disinfectant Towel with Bleach
Hype-Wipe® Bleach Towelette
SS
(Sample Descriptor)
W#
Decontamination wipe
For Task 1, # will be the number of coupons wiped
D#
Dry wipe
For Task 2, # will be the time in minutes after which the sample is collected
P(X)#
For Task 3, # will the 1 - slight pressure,, or 3 - higher pressure
PA for person A, PB for person B
N
(Replicate Number)
N
Sequential numbers
3.6.6 Phase 2 Sample Identification
Each sample was assigned a unique ID based on the sample coding outlined in Table 3-2. The sampling
team maintained an explicit laboratory log which included records of each unique sample number and its
associated test number, contamination application, sampling method, and the date sampled. Each coupon
was marked with only the material descriptor and unique code number. Once samples were transferred to
the NHSRC Biocontaminant Laboratory (Biolab) for plate counting, each sample was further identified by
replicate number and dilution factor. The NHSRC Biolab also included on each plate the date it was placed in
the incubator.
20
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Table 3-2: Sample Coding for Phase 2
Coupon Identification: 73-TID-L-M-D-SS-N
Category
Example Code
Description
TID
(Test ID)
1C
ForTask 1*: T=1, ID = C (Clorox®wipe) or H (Hype-Wipe®)
L
(Inoculation Location)
a
a = the upper left corner, e = the center, i = the lower right corner, 0 =
broad area; see Figure 3-2
M
(Material)
G
G = Glass
D = Painted Drywall
D
Decontamination Application
2
For Taskl:
1 = Slight Pressure
2 = Higher Pressure
SS
(Sample descriptor)
T(^ - Wipe samples from test
coupons
XT for blank coupon (negative)
# represents section the test sample was obtained from (a-i)
D
Drip sample
(X)M
MDI control sample from stainless steel coupons; XM for Blank Sample
SW(#)
Swab sample- followed by identifier (ty of where sample was collected
(A)-ADA
(C) - Coupon
P(ty - Wipe samples from
Positive Control coupons
# represents section the test sample was obtained from (a-i)
N
(Replicate Number)
1
Sequential test numbers
NHSRC Biolab Plate Identification: 73-TID-L-M-D-SS-N -R-D
73-TID-L-M-D-SS-N
As above
R (Replicate)
R
A-C
D (Dilution)
1
0 to 4, for 10E0 to 10E4
* Original study included Task 2 related to use of robots for decontamination. Only outcome from Task 1 is reported here.
3.6.7 Sample Frequency
Table 3-3 lists the sample frequency for decontamination tests using the wiping approach.
21
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Table 3-3. Wiping Efficacy - Sample Frequency
Sample Type
Quantity
Frequency
Location/Condition
Purpose
Test Coupon
Samples
Three coupon replicates per
test; nine wipe samples per
coupon for a total of 27 samples
One set per
material
Decontaminated
To determine the number of
viable spores after
decontamination
Procedural Blank
One coupon of equal size as
test coupon, not inoculated
One per
material
Decontaminated
To determine extent of cross
contamination during testing
Negative Control
Coupon
One 14 in x 14 in coupon
One per
material
Not decontaminated
To determine extent of cross
contamination and/or the
sterility of coupons
Task 1 Positive
Control Coupon
Two coupons per material,
inoculated as the first and last
coupon during the inoculation
procedure; nine wipe samples
per coupon for a total of 18
wipe samples
One set per
material
Not decontaminated
To determine the number of
viable spores recoverable from
the coupons
MDI Control Coupons
(stainless steel)
Three per inoculation event, as
the first and last inoculations.
Also includes the middle
inoculation if more than 25
inoculations are performed
Three per
inoculation
Not decontaminated
To determine the number of
viable spores deposited onto
the coupons and to assess the
stability of the MDI
Laboratory Material
Blanks
Three per material
Once per use
of material
NA
To demonstrate sterility of
extraction and plating materials
ADAs Sterility Check
(swab samples)
One per sterilized batch of
ADAs
Once before
testing
NA
To demonstrate sterility of ADAs
Drip Wipe
Three per coupon
Three
composite
samples per
test
Underneath the
coupon
Determine the transport of
spores off decontaminated
surface through drips
Decontamination
Wipe
One per coupon
Three per test
The decontamination
wipe itself
Determine the survival rates of
the spores on the wipe and the
physical removal of viable
spores after 24-h hold time
22
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3.6.8 Prevention of Cross Contamination of Samples during Sampling
Sampling posed a potentially significant opportunity for cross contamination of samples. To minimize the
possibility of cross contamination, several management controls were implemented:
• In accordance with aseptic techniques, a three-person sampling team was utilized, including a "lead
sampler," an "assistant sampler," and a "coupon handler."
• The "coupon handler" was designated as the only person to operate the ADA and handle the test
coupons during the sampling event.
• The "lead sampler" handled only the sampling media (wipes and liquids) and performed the surface
sampling of the test coupons.
• The "assistant sampler" was responsible for and handled only the containers for receiving the
sample collection media (e.g., a PRB wipe). The "lead sampler" deposited the collection media into
the sample containers that were opened, held, and sealed by the "assistant sampler." Sealed
samples were handled only by the "assistant sampler" who was solely designated to perform the
following actions using aseptic technique:
o The sealed bag with the sample was placed into another sterile plastic bag that was then
sealed; the exterior of that bag was then decontaminated using a bleach wipe.
o The double-bagged sample was placed into a sample container for transport.
o The exterior of the transport container was decontaminated by wiping all surfaces with a
bleach wipe or a towelette moistened with a solution of hypochlorite prior to transport from
the sampling location to the on-site NHSRC Biolab.
• After the sample was placed into the container for transport, the "coupon handler" placed the
sampled test coupons in pAB for decontamination prior to reuse or disposal.
• At the completion of each sampling event, each member of the sampling team changed their gloves
in preparation for working with the next sample.
As a further precaution to avoid cross contamination of samples, the order of coupon sampling was from
coupons least contaminated with B. atrophaeus to those most contaminated; i.e., (1) all blank coupons, (2) all
decontaminated coupons, and (3) positive control coupons.
Strict adherence to aseptic laboratory technique was followed by the NHSRC Biolab to recover, plate,
culture, and analyze samples. The order of analysis was the same as the order of sampling; i.e., (1) all blank
coupons, (2) all decontaminated coupons, and (3) all positive control coupons.
3.6.9 Collecting Representative Samples
The representativeness and uniformity of test coupon materials was considered essential in achieving
defensible evaluation results. Material representativeness was achieved by using materials that were typical
of those currently used in buildings in terms of quality, surface characteristics, and structural integrity and
which conformed to industry standards or specifications for indoor use. Material uniformity was achieved by
obtaining and preparing a quantity of material sufficient to allow multiple test samples to be prepared with
presumably uniform characteristics.
23
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3.6.10 Sample Storage and Preservation
After sample collection, sample integrity was accomplished by triple containment of samples: (1) sample
collection container, (2) sterile bag with exterior sterilized during the sample packaging process, and (3) clean
container holding all samples from a test. All individual sample containers remained sealed while in the
coupon decontamination laboratory and while in transport to the NHSRC Biolab. The sampling person did not
handle any samples after they were relinquished to the support person during placement into the primary
sample container.
All samples received were stored in a refrigerator at 4 °C ± 2 °C until they were analyzed. All samples were
allowed to stabilize at room temperature for one h prior to analysis.
3.6.11 Sample Holding Time
After sample collection for a single test was completed, all biological samples taken for that test were
transported to the NHSRC Biolab immediately along with the appropriate chain of custody (CoC) form(s). The
quality assurance project plan (QAPP) for this project stated that samples were to be stored no longer than
five days before commencing the primary analysis. However, given the volume of samples generated over a
short period of time, the samples were stored in the refrigerator for longer than five days before commencing
the primary analysis. A method development test was conducted to determine if the holding times had a
negative effect on the samples. No significant change was observed in the samples and bacterial colony
counts. These results are explained in detail in Section 5.3.2.
3.6.12 Sample Chain of Custody
Test schedules were coordinated with the NHSRC Biolab so that samples were collected, transferred, and
analyzed within the established timeframe, with the exception of hold times as discussed above. To ensure
the integrity of samples and to maintain a timely and traceable transfer of samples, an established and
proven CoC approach was used. The primary objective of the CoC was to create an accurate written record
that could be used to trace the possession of a sample from the moment of its creation through the reporting
of the results. A sample was said to be in custody if it was in any one of the following states:
• In actual physical possession
• In view, after being in physical possession
• In physical possession and locked up so that no one could tamper with it
• In a secured area, restricted except to authorized personnel
• In transit.
Test team members received copies of the test plans prior to each test. Pre-test briefings were then held to
apprise all participants of the objectives, test protocols, and CoC procedures to be followed.
In the transfer of custody, each custodian signed, recorded, and dated the transfer. Sample transfer was
either on a sample-by-sample basis or on a bulk basis. A CoC record listing the samples by their IDs always
accompanied the samples at each stage of their journey from initial collection through analysis and final
disposition. When turning over possession of samples, the transferor and recipient both signed, dated, and
24
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noted the time on the CoC record sheet. The recipient of the samples was declared to be the custodian and
was responsible for the samples until a subsequent transfer or disposition was made.
3.6.13 Sample Archiving
All samples and diluted samples were archived for at least two weeks following completion of analysis. This
archival period allowed for review of the data to determine if any re-plating of selected samples was required.
Samples were archived by maintaining the primary extract at 4 °C ± 2 °C in a sealed extraction tube.
3.6.14 Microbiological Analysis
The NHSRC Biolab located at the EPA facility in Research Triangle Park, NC, analyzed samples either
qualitatively for spore presence (quality control, swab samples) or quantitatively for the number of viable
spores recovered per sample (CFU). Details of the analysis procedures are provided below. A laboratory
notebook was used to document the details of each sampling event (or test).
Spores were extracted from the PRB wipes by adding 20 mL PBST to each tube, then agitating the tubes
using a vortex mixer (set to maximum rotation) for two minutes in 10-second intervals. For all sample types,
liquid extracts were serially diluted tenfold (in PBST) and 0.1 mL spread-plated in triplicate onto tryptic soy
agar (TSA) plates. Plates were incubated at 35 °C ± 2 °C for 18 to 24 h, and CFU were enumerated visually.
Only plates containing between 30 and 300 CFU were utilized for recovery estimates. Extracts were diluted
and replated if none of the tenfold dilutions resulted in all three plates containing colony counts within the
acceptable range. All extracts were stored at 4 °C ± 2 °C. Total spore recovery was calculated by multiplying
the mean CFU counts from triplicate plates by the inverse of the volume plated, by the dilution factor, and
finally by the total volume of the extract. Any samples below countable criteria (30-300 CFU) on the primary
dilution plates were subsequently filter plated through 0.2 jjm pore-size filters (Nalgene, Rochester, NY), with
the filters placed onto TSA plates followed by incubation at 35 °C ± 2°C for 18-24 h. The CFU counts from
these plates were used to calculate recovery in these circumstances.
Prior to erecting a coupon on the easel, the easel tray was lined with sterile drip wipes (Fisherbrand™ Dry
Clean-Wipes™, catalog no. 06-664-14, Pittsburgh, PA). The drip wipes collected any runoff/spores that were
displaced from the coupons during the decontamination wiping process. After the coupons were sampled,
the drip wipes were aseptically transferred to a sterile specimen cup and stored in the refrigerator at 4 °C for
a minimum of 24 h, after which they were neutralized by adding 70 mL of PBST with 2.5% STS.
The decontamination wipes were also stored in sterile specimen cups following the decontamination
procedure. They were refrigerated at 4 °C for a minimum of 24 h after which they were neutralized by adding
35 mL of PBST with 2.5% STS. The drip wipes as well as the decontamination wipes followed the same
extraction and microbial analysis procedure as the PRB sampling wipes.
25
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4 Results and Discussion
4.1 PHASE 1: Operational Aspects of Decontamination by Sporicidal Wipes
Results
Appendix A provides the tabulated results of Tasks 1, 2, and 3 of the preliminary study designed to establish
the wiping techniques and to aid in the selection of the decontamination wipes to be used in Phase 2 to
evaluate the effectiveness of decontamination methods. The conclusions from these test results are
explained in detail in the following sections.
4.1.1 Determination of Effective Surface Coverage of Sporicidal Wipes
The objective of this task was to determine how much liquid was dispensed from a single wipe. In this task,
the weight loss of a wet wipe following the prescribed wiping procedure was taken to be equal to the amount
of bleach deposited on a coupon. The individually packaged, single-use Hype-Wipe® Bleach Towelettes
resulted in higher amount of bleach dispensed, for all types of materials tested, compared to the other wipes
that are susceptible to drying out faster once their canister lid is opened. The weight losses were the highest
for the largest areas wiped. As expected, the amount of dispensed liquid bleach per unit area decreased with
increasing areas wiped as illustrated in Figures 4-1 and 4-2 for all material/wipe combinations tested. Among
the four materials tested, the least amount of liquid bleach was transferred to glass, while painted drywall
retained the most liquid (Figure 4-3). Since a surface area of 9 ft2 resulted in 40 to 50% of the bleach being
dispensed regardless of material/wipe type tested, glass (nonporous) and painted drywall (porous) were
selected as the materials to be tested during the Phase 2 decontamination testing.
4.0.
3 3.0-
8 2.5:
^ 2.0-|
15-1
fo.5-1
CD 0.0 ¦
c 4.0'
P 3 5 "I
g- 3.0 -
d 2.5-
-i 2.0:|
01
Melamine
Hype-VWpe® Bleach Towelette
Clorox® Bleach Germicidal Wpe
Sani-Cloth® Bleach Germicidal VWpe
Dispatch® Towel with Bleach
1.0-
0.5-^
0.0
Glass
1 x1 1.5x1.5 2x2 2.5x2.5 3x3
Coupon Size (ft x ft)
Figure 4-1. Residual Bleach Recovered from Melamine and Glass Material
26
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c^20-|
15-
¦
Glass
•
Melamine
A
Vinyl Flooring
~
Painted Drywal
*
~
*
T
I
1
1x1 1.5x1.5 2x2 2.5x2.5
Coupon Size (ft x ft)
3x3
Figure 4-3. Percent Liquid Dispensed on Material as a Function of Material Type, for Hype-Wipe®
27
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4.1.2 Surface Retention Times of Wipe Decontaminant Liquid Results
This task was designed to assess the propensity of each material to retain the liquid bleach from each wipe
type on its surface overtime and to assess the temporal liquid loss as a function of the material/wipe
combination. Two sets of five replicate 1 ft x 1 ft coupons for each material were used for this task. The
amount of decontamination liquid dispensed was obtained for each different type of wipe by the same
method used in Task 1. The amount of liquid remaining on a coupon immediately after the decontamination
event (time zero) and after a five-minute drying time was determined by dry wiping of the surface. The liquid
remaining on the surface was equated with the increase in weight of the dry wipes at the two time points (0,
and five (5) minutes). The time point of five minutes was selected as the longest minimum contact time,
across the four different sporicidal wipes, that the decontamination liquid was supposed to have with the
material to achieve the efficacy criteria as demonstrated on a 1 ft x 1 ft coupon by Meyer et al. [1], The results
for the temporal recovery of the bleach solution for each type of material/wipe type combination are
presented in Figure 4-4 for Clorox® Bleach Wipe and Hype-Wipe® Bleach and in Figure 4-5 for Sani-Cloth®
Bleach Germicidal Disposable Wipe and Dispatch® Hospital Cleaner Disinfectant Towel with Bleach,
respectively.
The residual liquid solution from the Hype-Wipe® Bleach Towelettes, and to a lesser extent the liquid solution
from the Clorox Healthcare® Bleach Germicidal Wipe, was found to adhere better to the materials tested and
was less susceptible to loss through evaporation or absorption through the material surface over time. The
residual liquid solution from the Sani-Cloth® Bleach Germicidal Disposable Wipe was found to disappear the
most through evaporation or absorption through the materials. Among the materials tested, the vinyl flooring
material was associated with the lowest loss in liquid overtime, while painted drywall was the most affected.
Although, all the tested wipes were saturated with 0.525% sodium hypochlorite (bleach), the contact time
between the bleach and a specific surface may be affected by the secondary active ingredients such as
alcohols, surfactants, and stabilizers.
28
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~o
0
if)
£=
0
Q.
(0
Q
JZ
o
03
_0
m
"D
'=3
O"
Zl
3.0
2.5-
2.0-
1.5-
1.0-
0.5-
0.0
3.0
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2.0-
1.5-
1.0-
0.5-
0.0
Gorox®Bleach Germicidal Wipe]
Jjn
I nomin
I BSmin
n~i
^—i—
Hype-Wipe® Beach Towelette
Glass Vinyl Flooring Melamine Painted Drywall
Material Type
Figure 4-4. Temporal Residual Bleach Recovered following a Decontamination Event using Clorox®
Bleach Wipe and Hype-Wipe® Bleach Towelette
1
"O
-------
4.1.3 Impact of Wiping Pressure
This task was to determine the effect of the application pressure on the decontamination wipe while wiping
the coupons. Coupons (1 ft x 1 ft) of each material were used for this task, and results are shown in Figures
4-6 and 4-7. Higher pressure application resulted in a higher volume of liquid solution dispensed compared to
the slight (low) pressure application for all material/wipe type combinations. For these particular tests, the
effect of applying higher pressure, compared to slight pressure, resulted in an increase of approximately 60%
of the liquid dispensed from the Hype-Wipe® Bleach Towelettes to the coupon and up to 300% from Sani-
Cloth® wipes. The decontamination efficiency of a particular wipe type may be affected by the operational
aspect of the decontamination procedure such as the application pressure.
€
"O
CD
(/)
C
a)
Q.
C/5
Q
_c
o
03
0
QQ 0-
"a
6-
5-
4-
3-
2-
1 -
Clorox® Bleach Wipe
Slight Pressure
Heavy Pressure
O"
_i
4—
O
6-
5-
4-
3-
2-
1 -
0-
Hype-Wpe® Towelette
Glass Vinyl Flooring Melamine
Material Type
Painted Drywall
Figure 4-6. Effect of Application Pressure on Liquid Dispensed Using Clorox® Bleach Wipe and Hype-
Wipe® Bleach Towelette
30
-------
€
"O
CD
(/)
c
a)
Q.
a)
6-
5-
4-
3-
2-
1 -
QQ 0-
"a
Dispatch® Towel with Bleach
Slight Pressure
Heavy Pressure
—±—
1—H-l
1
O"
_i
4—
O
6-
5-
4-
3-
2-
1 -
0-
Sani-Cloth® Bleach Germicidal VMpe
Glass Vinyl Flooring Melamine Painted Drywall
Material Type
Figure 4-7. Effect of Application Pressure on Liquid Dispensed using Sani-Cloth® Wipe and
Dispatch® Towel with Bleach
4.2 PHASE 2: Evaluation of Efficacy of Hypochlorite Wipes Results
Appendix B provides the tabulated recoveries of the Phase 2 testing designed to evaluate the
decontamination effectiveness of targeted wipe/material type combinations to decontaminate a 12.25 ft2
surface. The variables evaluated in the complex test matrix included two types of decontamination wipes
(Hype-Wipe® Bleach Towelette, and Clorox Healthcare® Wipe), two material types (glass and painted
drywall), two inoculation spatial distributions (Hot Spot versus Broad Area), and the effect of wipe application
pressure (slight and higher pressure). In addition, the fate of the spores is presented for different inoculation
locations (center (e), the upper left corner (a), and the lower right corner (i) for each Hot Spot inoculation, as
shown in Figure 3-3), and for the broad area inoculation.
The two different hypochlorite wipes were tested on 3.5 ft x 3.5 ft (107 cm x 107 cm) coupons, consisting of 3
x 3 sections of 14 in x 14 in (35.6 cm x 35.6 cm) sampling areas. Each 14 in x 14 in area was sampled
individually after a 30-min contact time following the wiping of the whole surface. Surfaces were wet with
locally dryer areas due to, e.g., the gravitational pull of the residual dispensed liquid toward the bottom of the
panel. Recovered spores (CFU) for each section were summed to determine the CFU and log CFU reduction
per coupon type/wipe type combination across the whole 42 in x 42 in area. The decontamination efficacy
results are shown in Tables 4-1 and 4-2 for glass and painted drywall, respectively. Some (3 out of 24) of the
procedural blanks had more than 500 spores. For those three test conditions, a 6 log reduction would be
impossible to demonstrate. It should be noted that the average recovered number of spores from test
31
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coupons were significantly higher than those from the procedural blank suggesting a less than 6 log
reduction. See Section 5.3.1 for a more detailed discussion.
One-way analysis of variance (one-way ANOVA), revealed that, at the 95% confidence interval (CI), the
mean decontamination efficiencies of the types of wipes used in this study are statistically different (p value =
0.031). Hype-Wipe® Bleach Towelettes with an overall average spore LR of 3.8 ± 0.8 were, in general, more
effective than the Clorox Healthcare® Wipe with an overall average spore LR of 3.0 ± 1.0, independent of the
inoculation location. Similar work performed by Meyer et al. [1] showed that these selected wipes completely
inactivated Bacillus spores (LR > 6.0) on smaller 14 in x 14 in (35.6 cm x35.6 cm) coupons for similar wipe
type/material combination decontamination approaches. Mean decontamination efficacies were not
statistically different across the two materials (p values = 0.436 and 0.294 for Clorox Healthcare® Wipe and
Hype-Wipe® Bleach Towelettes, respectively). The wipe application pressure seems to have little or no
effect on the decontamination efficacy of the wipe. No detailed statistical analysis was performed on this
small subset of test samples.
To interpret cross contamination of the wipes during the decontamination process, the spatial distribution
of the post-decontamination spore concentration for both the Hype-Wipe® Bleach Towelette and Clorox
Healthcare® Wipe on glass and painted drywall material is illustrated in Figures 4-8 to 4-11 using a color-
coded scheme for the spore loading concentration. The results show clearly that the wipe decontamination
was most effective when the Hot Spot inoculation occurred in the upper left corner, which is the same
location where the (horizontal motion) decontamination wiping started, and worst for the Hot Spot located
in the lower right corner of the coupon (where the decontamination wiping ended). For the lower right
corner inoculation test, the first surface area of the hypochlorite wipe used to decontaminate the coupon
using horizontal motion (working downward to completely cover the surface) may have dried before
reaching this Hot Spot. The dried wipe may have then become contaminated without inactivation of
spores. The cross contamination between the wipe surfaces (spent dry wipe surface folded inwards and
newly saturated wipe surface) may also have occurred during the folding of the wipe, and spores were
subsequently distributed throughout the other areas of the coupon during the vertical wipe
decontamination pathway. The third (diagonal wiping) and the fourth (perimeter wiping) pathways may
have exacerbated the cross contamination, due to faster drying of the exposed surface, since only one
fourth and one eighth of the original surface area were used during these two last steps.
Residual spores collected from sterile drip Clean-Wipes™ that lined the easel tray supporting the test coupon
and stored for a minimum of 24 h at 4 °C in sterile specimen cups showed an overall contamination of less
than 0.03% of the inoculated spore burden. This contamination may be credited to liquid runoff during the
decontamination wiping procedure and/or re-aerosolization/deposition of spores during the coupon
setup/decontamination/sampling events. Enumeration of CFU on the decontamination wipes that were used
during the decontamination process and stored for a minimum of 24 h at 4 °C in sterile specimen cups
resulted in no spores being detected. This indicates that spores collected on the sporicidal wipe became
inactivated.
32
-------
Table 4-1. Decontamination Results for Glass Material Type
Decontamination
Wipe Type
Inoculation
Location
Applied
Sampling
Pressure
Average Recovery (CFU)
Log
Reduction
Positive Control
Test Coupons
Hype-Wipe®
Bleach
Towelette
Upper left
corner (a)
Slight
3.17 x 107 ± 2.19 x 107
2.60 x 102 ± 1.10 x 102
5.12 ±0.88
Center (e)
Slight1
4.43 x 107± 8.50 x 106
2.26 x 104 ± 1.20 x 104
3.86 ± 0.91
Lower right
corner (i)
Slight
4.35 x 107 ± 9.66 x106
1.72 x 107 ± 1.14 x 104
3.60 ± 0.56
Heavy
3.90 x 107 ± 2.83x106
1.38 x 103 ± 9.94 x 102
4.63 ±0.91
Broad Area
Slight
6.08 x 106 ± 1.24 x 106
4.33 x 102 ± 5.53 x102
4.52 ±1.42
Heavy
7.91 x106± 1.74 x106
3.49 x 103 ± 4.58 x103
3.88 ±1.26
Clorox
Healthcare®
Bleach
Germicidal Wipe
Upper left
corner (a)
Slight
3.06 x 107 ± 9.68 x 106
3.32 x 103 ± 4.21 x103
4.32 ± 0.94
Center (e)
Slight
5.04 x 107 ± 1.27 x 107
2.84 x 106 ± 1.41 x106
1.91 ±0.42
Lower right
corner (i)
Slight
2.14 x 107 ± 1.31 x 107
1.61 x 103 ± 2.12 x 103
4.50 ±1.20
Heavy
4.83 x 107 ± 8.74 x106
1.70 x 104 ± 9.18 x 104
3.50 ±0.21
Broad Area
Slight
7.11 x106± 1.44 x106
1.21 x104± 6.21 x103
2.86 ± 0.27
Heavy
6.55 x 106 ± 9.02 x10s
3.34 x 104 ± 1.08 x 103
3.31 ±0.14
1 Procedural blank showed relatively high contamination for these tests (>5 x 102 spores)
33
-------
Table 4-2. Decontamination Results for Painted Drywall Material Type
Decontamination
Wipe Type
Inoculation
Location
Applied
Sampling
Pressure
Average Recovery (CFU)
LR
Positive Control
Test Coupons
Hype-Wipe®
Bleach Towelette
Upper left
corner (a)
Slight
4.54 x 107 ± 1.24 x 107
3.25 x 103 ± 4.36 x103
4.63 ±1.33
Center (e)
Slight
6.11 x 107 ± 1.45x 107
5.54 x 104± 2.85 x104
3.47 ± 0.79
lower right
corner (i)
Slight
5.03x 107 ± 3.11 x 107
2.04 x 104 ± 1.53 x 104
3.77 ± 0.86
Heavy1
3.76 x 107 ± 1.27 x106
4.75x 105 ± 2.14x10s
2.20 ±0.26
Broad Area
Slight1
7.22 x 106 ± 1.70 x106
7.77 x 103 ± 4.27 x103
3.02 ± 0.23
Heavy
1.06 x 107 ± 6.65x 105
8.37 x 103 ± 5.07 x103
3.22 ± 0.39
Clorox
Healthcare®
Bleach Germicidal
Wipe
Upper left
corner (a)
Slight
5.38 x 107 ± 6.46 x106
1.00 x 104 ± 1.35 x 104
4.24 ±1.08
Center (e)
Slight
7.52 x 107 ± 1.97 x107
1.77 x 105 ± 5.53x104
2.73 ±0.23
lower right
corner (i)
Slight
4.43 x 107 ± 9.52 x106
1.23 x 106 ± 5.31 x 105
1.64 ±0.11
Heavy
3.67 x 107 ± 7.35 x106
2.78 x 105 ± 3.44 x 105
2.46 ± 0.32
Broad Area
Slight
9.87 x 106 ± 5.66 x 105
8.13 x104± 2.79 x104
2.20 ±0.18
Heavy
9.39 x 106 ± 2.60 x106
1.09 x 105 ± 2.8 x 104
1.95 ±0.08
1 Procedural blanks show relatively high contamination for these tests (>5 x 102 spores)
34
-------
H1GI
H2GI
H3GI
HOGI
3.0E+01
ND
2.1E+01
3.0E+02
ND
7.7E+01
ND
ND
ND
2.9E+01
1.4E+00
5.8E+03
ND
1.1E+02
6.9E+03
ND
7.3E+01
5.1E+04
1.0E+02
1.6E+03
2.0E+01
1.2E+02
1.2E+03
3.9E+02
4.4E+03
1.5E+02
1.7E+04
9.0E+01
ND
ND
ND
ND
ND
1.3E+00
1.5E+01
2.6E+00
2.6E+02
ND
1.3E+01
ND
ND
5.4E-01
1.1E+00
ND
ND
2.6E+00
ND
6.5E+02
ND
1.3E+00
1.9E+02
ND
4.0E+00
1.2E+03
1.4E+01
1.1E+02
1.2E+01
1.2E+02
3.6E+02
4.0E+01
2.9E+00
1.4E+03
4.2E+00
1.4E+02
ND
ND
3.9E+02
ND
ND
3.4E+02
2.7E+02
ND
1.7E+01
ND
ND
ND
ND
5.6E+01
ND
ND
ND
ND
ND
ND
ND
ND
1.4E+00
2.2E+02
6.4E+01
1.4E+03
ND
ND
ND
ND
ND
ND
ND
2.6E+00
2.4E+04
1.3E+01
ND
1.1E+00
ND
ND
ND
1.3E+01
1.6E+01
2.5E+00
4.7E+07
1.3E+05
2.4E+03
2.3E+05
3.4E+03
6.8E+00
1.4E+01
5.5E+00
2.7E+02
3.1E+01
2.4E+03
ND
1.9E+03
5.0E+07
7.3E+03
ND
7.7E+03
6.3E+01
NA
ND
ND
ND
1.3E+01
6.6E+02
ND
5.4E+00
3.7E+07
1.2E+06
5.8E+05
5.1E+05
6.7E+05
5.2E+05
3.9E+05
7.3E+05
5.9E+05
1.8E+06
1.6E+07
2.8E+04
5.3E+03
2.0E+05
1.8E+03
1.1E+03
3.7E+01
1.9E+01
ND
ND
2.3E+02
8.1E+01
3.9E+03
3.8E+07
6.0E+02
1.8E+01
2.0E+01
1.2E+02
1.3E+00
ND
ND
1.7E+01
1.3E+00
1.3E+00
6.5E+00
9.4E+01
5.0E+07
6.2E+05
5.6E+05
5.1E+05
3.6E+05
5.3E+05
6.0E+05
9.2E+05
4.7E+05
6.4E+05
Coupon 1 Coupon 2 Coupon 3 Positive Control 1 Positive Control 2
Figure 4-8. Spatial Post-Decontamination Residual Spore Concentration Using Hype-Wipe® Bleach Towelette /Glass Material at Different
Inoculation Locations.
Spore loading concentration color coded scheme; ND: no viable spores detected; NA: Not Available due to sample loss.
35
-------
C1GI
C2G1
C3GI
COGI
4.9E+01
1.1E+00
5.1E+02
ND
7.5E+01
ND
4.4E+00
3.2E+01
2.2E+00
3.9E+05
4.3E+05
5.0E+05
5.0E+05
1.5E+06
1.7E+06
4.8E+05
9.9E+05
5.7E+05
ND
ND
7.4E+01
ND
ND
1.3E+01
1.5E+01
1.3E+00
1.5E+02
2.3E+01
1.8E+02
1.6E+02
1.5E+01
4.1E+01
6.2E+01
6.7E+02
4.2E+02
2.4E+03
8.6E+03
1.1E+00
3.5E+01
1.4E+02
ND
1.1E+00
ND
ND
ND
8.6E+03
9.0E+01
3.5E+01
4.5E+02
2.9E+02
2.0E+03
6.1E+03
4.4E+03
1.5E+03
2.4E+03
2.2E+02
7.6E+00
4.3E+02
6.3E+01
ND
1.2E+03
5.6E+01
1.2E+01
1.7E+01
2.4E+00
2.6E+02
1.1E+02
2.9E+02
8.6E+02
4.1E+03
3.0E+03
1.8E+03
4.3E+00
ND
ND
ND
5.4E+01
2.2E+00
2.6E+02
9.2E+01
9.0E+01
1.2E+04
3.8E+04
3.0E+04
2.8E+04
4.7E+04
2.6E+05
4.6E+04
4.0E+04
8.9E+05
2.7E+01
ND
2.5E+00
ND
ND
9.2E+01
4.8E+01
ND
4.0E+01
1.2E+01
5.8E-01
4.7E+02
ND
1.1E+00
2.8E+01
9.7E+02
2.0E+04
6.2E+02
2.4E+07
3.9E+04
2.2E+03
1.9E+04
1.7E+03
2.2E+03
1.5E+01
3.7E+01
2.2E+00
5.8E+00
1.6E+03
ND
1.7E+03
4.1E+07
7.7E+02
3.3E+02
3.7E+02
5.9E+00
2.7E+00
1.3E+00
ND
1.2E+00
ND
2.5E+01
2.5E+00
7.7E+01
1.22E+07
9.8E+05
1.1E+06
7.1E+05
1.3E+06
8.0E+05
8.9E+05
5.7E+05
8.5E+05
7.9E+05
3.7E+07
1.3E+05
4.6E+02
2.8E+05
3.8E+03
2.2E+00
1.5E+03
1.1E+00
1.0E+01
1.3E+01
8.5E+02
1.5E+01
1.0E+04
5.9E+07
3.2E+04
2.9E+01
7.4E+02
5.3E+02
3.8E+00
2.5E+00
1.27E+00
4.3E+01
5.9E+00
7.4E+03
5.3E+00
2.1E+02
3.07E+07
1.3E+06
7.3E+05
8.5E+05
1.1E+06
8.5E+05
8.1E+05
8.3E+05
9.9E+05
7.1E+05
Coupon 1
Coupon 2
Coupon 3
Positive Control 1
Positive Control 2
Figure 4-9. Spatial Post-Decontamination Residual Spore Concentration using Clorox Healthcare® Wipe/Glass Material at Different Inoculation
Locations
EO
El
E2
E3
E4
E5
E6
Spore loading concentration color coded scheme; ND: no viable spores detected.
36
-------
H1DI
H2DI
H3DI
HODI
6.0E+02
ND
ND
4.2E+01
7.6E+01
3.7E+01
3.0E+01
1.1E+01
1.2E+01
5.2E+01
1.6E+03
4.8E+02
1.6E+04
8.9E+04
6.2E+02
1.5E+03
1.1E+04
2.8E+02
ND
1.3E+00
ND
ND
2.1E+01
4.7E+02
2.7E+01
2.9E+01
3.0E+02
8.2E+00
1.5E+02
5.2E+02
5.5E+02
6.5E+02
2.4E+03
1.2E+03
2.1E+03
7.7E+02
8.6E+03
2.6E+00
1.3E+00
1.1E+01
1.4E+02
1.6E+01
7.8E+00
5.2E+00
1.3E+00
1.4E+00
2.8E+00
1.2E+01
5.5E+01
1.2E+03
7.5E+01
9.8E+01
8.4E+01
7.0E+01
ND
ND
ND
3.8E+03
ND
3.6E+01
1.7E+01
1.5E+03
1.1E+04
8.6E+03
1.9E+01
2.1E+01
9.5E+01
1.4E+01
7.9E+01
1.4E+03
1.0E+03
5.0E+02
ND
8.4E+00
4.4E+01
3.0E+01
ND
5.7E-01
1.4E+00
7.1E+01
7.5E+00
ND
1.4E+01
2.3E+01
1.7E+03
4.2E+04
7.8E+02
4.0E+01
3.3E+02
3.1E+02
ND
ND
ND
1.3E+00
ND
3.6E+02
6.5E+02
2.6E+02
4.3E+04
2.5E+01
1.5E+02
1.3E+02
7.2E+01
3.7E+02
4.6E+01
1.4E+03
3.3E+02
7.0E+02
5.4E+07
2.3E+02
4.5E+00
3.7E+02
1.1E+00
3.3E+00
ND
ND
1.2E+01
5.6E+00
4.3E+02
ND
4.0E+04
5.1E+07
2.8E+03
3.3E+02
1.5E+03
ND
1.0E+01
1.0E+01
1.0E+01
ND
1.0E+01
1.1E+02
ND
ND
4.8E+07
9.1E+05
1.1E+06
7.6E+05
7.5E+05
1.0E+06
8.9E+05
1.1E+06
8.9E+05
4.7E+05
3.7E+07
1.1E+00
1.1E+00
1.7E+03
3.8E+03
ND
6.0E+01
1.1E+00
ND
1.1E+02
4.1E+03
1.1E+01
1.7E+04
7.1E+07
2.1E+04
4.9E+02
8.8E+03
1.6E+02
7.8E+00
2.0E+01
ND
9.0E+02
6.3E+01
2.3E+01
2.3E+01
5.6E+02
5.3E+07
9.7E+05
9.1E+05
8.6E+05
1.3E+06
1.0E+06
7.5E+05
9.0E+05
8.4E+05
8.6E+05
Coupon 1 Coupon 2 Coupon 3 Positive Control 1 Positive Control 2
Figure 4-10. Spatial Post-Decontamination Residual Spore Concentration using Hype-Wipe® Bleach Towelette /Painted Drywall Material at
Different Inoculation Locations
Spore loading concentration color coded scheme; ND: no viable spores detected.
37
-------
C1DI
C2DI
C3DI
CODI
4.7E+02
ND
1.7E+03
ND
1.1E+00
5.1E+01
7.1E+01
3.3E+00
6.3E+01
3.4E+02
1.1E+02
1.5E+03
9.8E+03
9.6E+03
7.5E+03
7.5E+03
2.3E+03
7.7E+03
1.5E+04
7.0E+03
3.3E+04
4.6E+03
5.7E+03
2.6E+04
9.4E+04
6.7E+04
1.7E+06
6.5E+03
1.4E+04
4.2E+03
3.2E+04
2.7E+03
1.9E+04
7.2E+03
2.6E+04
4.7E+04
2.6E+04
1.3E+01
9.1E+02
1.2E+00
2.6E+02
2.0E+02
2.8E+01
1.5E+02
2.2E+01
8.6E+03
1.5E+02
3.7E+02
1.1E+04
1.9E+05
4.7E+04
4.0E+03
4.5E+03
2.0E+04
2.1E+04
9.2E+03
1.2E+04
1.1E+04
4.5E+03
9.5E+03
5.5E+04
8.4E+04
1.1E+06
6.7E+02
4.3E+03
2.4E+03
3.9E+03
6.1E+03
3.2E+04
2.5E+03
3.3E+03
4.0E+03
2.9E+01
1.7E+01
2.8E+02
1.8E+01
1.1E+00
9.5E+01
7.6E+00
5.5E+00
1.6E+01
1.1E+01
3.2E+02
3.4E+03
1.2E+04
1.3E+05
4.0E+04
2.3E+03
6.7E+03
3.4E+03
3.5E+03
8.8E+02
3.9E+03
5.5E+03
NA
3.3E+03
2.0E+04
2.6E+04
3.0E+05
7.3E+01
3.5E+02
6.1E+01
6.5E+02
1.2E+04
1.8E+02
3.3E+03
5.0E+03
3.9E+03
4.9E+07
1.3E+03
1.2E+00
1.1E+02
1.2E+01
4.3E+00
1.0E+01
2.5E+00
3.6E+01
1.3E+04
2.0E+05
1.2E+01
1.1E+05
6.1E+07
5.7E+04
1.9E+04
3.0E+05
9.2E+03
4.2E+00
3.4E+02
1.4E+00
ND
1.4E+00
6.2E+02
3.2E+03
3.8E+02
5.1E+07
1.4E+06
1.5E+06
1.5E+06
1.1E+06
1.1E+06
1.6E+06
1.3E+06
1.4E+06
1.0E+06
5.8E+07
9.4E+02
3.3E+00
3.1E+04
3.8E+03
3.3E+02
1.2E+01
2.3E+01
ND
5.7E+03
1.0E+05
1.4E+00
2.8E+04
8.9E+07
2.0E+04
2.6E+02
2.2E+05
2.3E+02
ND
1.5E+01
ND
1.3E+00
3.8E+03
1.4E+00
2.8E+00
2.7E+00
3.8E+07
1.3E+06
1.1E+06
9.5E+05
1.0E+06
9.2E+05
1.1E+06
2.3E+06
7.8E+05
8.9E+05
Coupon 1
Coupon 2
Coupon 3
Positive Control 1
Positive Control 2
Figure 4-11. Spatial Post-Decontamination Residual Spore Concentration using Clorox Healthcare® Wipe/Painted Drywall Material at Different
Inoculation Locations
EO
El
E2
E3
E4
E5
E6
Spore loading concentration color coded scheme; ND: no viable spores detected; NA: Not Available due to sample loss.
38
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5 Quality Assurance and Quality Control
This project was performed under the approved Category III QAPPs entitled "Decontamination Solution
Methods for Bacillus anthracis Surrogates Sporicidal Wipes Product Assessment Part I", July, 2014, and
"Evaluation of Sporicidal Wipes and Liquid Agents for Decontamination of Anthrax-Contaminated Surfaces
by Hand and Robotic Cleaners", November, 2014.
All test activities were documented via narratives in laboratory notebooks and the use of digital photography.
The documentation included, but was not limited to, a record for each decontamination procedure, any
deviations from the QAPP, and physical impacts on materials. All tests were conducted in accordance with
developed Decontamination Technologies Research Laboratory (DTRL) and NHSRC Biolab Miscellaneous
Operating Procedures (MOPs) to ensure repeatability and adherence to the data quality validation criteria set
for this project.
5.1 Criteria for Critical Measurements/Parameters
The Data Quality Objectives (DQOs) are used to determine the critical measurements needed to address the
stated objectives and specify tolerable levels of potential error associated with simulating the prescribed
decontamination environments. The following measurements were deemed to be critical to accomplish part
or all of the project objectives:
• Weight of the decontamination and dry wipes
• Sample volume collected
• Plated volume
• Counts of CFU.
Data quality indicators (DQIs) for the critical measurements were used to determine if the collected data met
the quality assurance objectives. A list of these DQIs can be found in Table 5-1. Failure to provide a
measurement to meet these goals resulted in a rejection of results derived from the critical measurement. For
instance, if the plated volume of a sample was not known (i.e., was not 100% complete), then that sample
was declared invalid. If a collected sample was lost or did not meet the criteria for other reasons, then
another sample was collected to take its place.
39
-------
Table 5-1. Critical Measurement Criteria
Critical Measurement
Measurement Device
Accuracy
Precision
Detection Limit
Sample Volume
Serological Pipette
Subdivision 0.5
mL
±0.2 mL
±0.1 mL
Plated Volume
Pipet
±2%
±1%
NA
CFU/Plate
Counting
± 10%
(between two
counters)
±5
1 CFU
Weight
Scale
0.004 g
0.0001 g
NA
NA = not applicable
5.2 Quality Control Checks
Many quality assurance (QA)/quality control (QC) checks were used in this project to ensure that the data
collected meet all the critical measurements listed in Table 5-1. The measurements/parameters criteria were
set at the most stringent level that can routinely be achieved. The integrity of the sample during collection and
analysis was evaluated. Control samples and procedural blanks were included along with the test samples so
that well-controlled quantitative values were obtained. Background checks for the presence of bacterial
spores were included as part of the standard protocol. Replicate coupons were included for each set of test
conditions. Validated operating procedures using qualified, trained and experienced personnel were used to
ensure data collection consistency. When necessary, training sessions were conducted by knowledgeable
parties, and in-house practice runs were used to gain expertise and proficiency prior to initiating the research.
The quality control checks that were performed in this project are described in the following sections.
5.2.1 Integrity of Samples and Supplies
Samples were carefully maintained and preserved to ensure their integrity. Samples were stored away from
standards or other samples that could possibly cross contaminate them.
Supplies and consumables were acquired from reputable sources and were National Institute of Standards
and Technology (NIST)-traceable whenever possible. Supplies and consumables were examined for
evidence of tampering or damage upon receipt and prior to use, as appropriate. Supplies and consumables
showing evidence of tampering or damage were discarded and not used. All examinations were documented
and supplies were appropriately labeled. Project personnel carefully checked supplies and consumables prior
to use to verify that they met specified task quality objectives and did not exceed expiration dates. All pipettes
were calibrated yearly by an outside contractor (Calibrate, Inc.), incubation temperature was monitored using
NIST-traceable thermometers, and balances were calibrated yearly by the EPA Metrology Laboratory.
40
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5.2.2 NHRSC Biolab Control Checks
Quantitative standards do not exist for biological agents. Quantitative determinations of organisms in this
investigation did not involve the use of analytical measurement devices. Rather, the CFU were enumerated
manually and recorded. If the CFU count for bacterial growth did not fall within the target range, the sample
was either filtered or re-plated. For each set of results (per test), a second count was performed on 25
percent of the plates within the quantification range (plates with 30 - 300 CFU). All second counts were found
to be within 10 percent of the original count.
5.3 QA/QC Sample Acceptance Criteria
The acceptance criteria for the critical CFU measurements were set at the most stringent level that could be
achieved routinely. Positive controls and procedural blanks were included along with the test samples in the
experiments so that well-controlled quantitative values were obtained. Background checks were also
included as part of the standard protocol. Replicate coupons were included for each set of test conditions.
Further QC samples were collected and analyzed to check the ability of the NHSRC Biolab to culture the test
organism, as well as to demonstrate that materials used in this effort did not themselves contain spores. The
checks included:
• Negative control coupons: sterile coupons that underwent the same sampling process;
• Field blanks: transfer of sterile wipes from the sample tube to an empty conical tube at the
decontamination location;
• Laboratory blank coupons: sterile coupons not removed from NHSRC Biolab;
• Laboratory material coupons: includes all materials, individually, used by the NHSRC Biolab in
sample analysis; and
• Stainless steel positive control coupons: coupons inoculated but not fumigated.
Additional QA/QC objectives are shown in Table 5-2. These QA/QC objectives provide assurances against
cross contamination and other biases in the microbiological samples.
41
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Table 5-2. QA/QC Sample Acceptance Criteria
Sample Type
Purpose
Acceptance Criteria
Corrective Actions
Frequency
Negative Control
Coupons
Determine extent of
cross contamination in
test area
None
Values on test coupons of
the same order of
magnitude will be
considered to have resulted
from cross contamination
One per test
Field Blanks
Verify the sampling
wipes do not introduce
contamination into
samples
No detectable spores
Determine source of
contamination and remove
One per sample type per
test
Laboratory Blank
Coupons
Verity the sterility of
coupons following
autoclaving
No detectable spores
Determine source of
contamination and remove
One per test per coupon
type
Laboratory Material
Coupons
Verity the sterility of
materials used to
analyze viable spore
count
No detectable spores
Determine source of
contamination and remove
Three per material per test
Blank TSA Sterility
Control
(plate incubated, but
not inoculated)
Controls for sterility of
plates
No observed growth
following incubation
All plates are incubated
prior to use, so any
contaminated ones will be
discarded
Each plate
Positive Control
Coupons
Used to determine the
extent of inoculation on
the coupons
1 x106CFU, ±0.5 log
(Broad Area Inoculation) or
1 x107CFU, ±0.5 log
(Hot Spot Inoculation)
Outside target range:
discuss potential impact on
results with EPA WACOR;
correct loading procedure
for next test and repeat
depending on decided
impact
Two per coupon type.
Procedural Blank
Coupons
Determine cross
contamination during the
testing procedures
No detectable spores
Determine source of
contamination and remove
One for each test condition
Inoculation Control
Coupons
Used to determine drift in
the MDI
The CFU recovered from
the first coupon must be ±
0.5 log of the last coupon
Reject results and repeat
test
Two per inoculation
Replicate Plating of
Diluted Microbiological
Samples
Used to determine
variability in CFU counts
The reportable CFU of
triplicate plates must be
within 100%. Reportable
CFU are between 30 and
300 CFU per plate
Re-plate sample
Each sample
WACOR = Work Assignment Contracting Officer Representative.
42
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5.3.1 QA/QC Test Results Validation
The QA/QC control test results for the whole sampling campaign are shown in Table 5-3. All field blanks and
inoculum blanks were found to be non-detects. However, some of the negative control blanks (2 out of 24)
and some of the procedural blanks (3 out of 24) were found to have approximately 5 x 102 CFU per coupon.
The source of this contamination is unknown. For negative controls, the contamination may have occurred by
incomplete inactivation of spores from the materials by the VHP cycle and the subsequent reuse of such a
coupon in the next test. Procedural blanks may have become contaminated due to their presence in an area
with inoculated coupons present. Due to the high levels of spores in the negative control and procedural
blanks, this study cannot demonstrate a minimum of 6 LR antimicrobial efficacy for every test condition. For
the test conditions where cross contamination occurred, the number of spores recovered from especially
the hot spot inoculated test coupons were significantly higher than those from the associated procedural
blank still suggesting that the sporicidal wipes were unable to deactivate all spores. The vast majority of
sporicidal wipe tests with associated non-detects in procedural blank or negative control support the
general observation that a 6 log efficacy was not reached when wiping a 3.5 ft x 3.5 ft surface area.
Table 5-3. QA/QC Test Results
Decontamination
Wipe Type
Inoculation
Location
Applied
Sampling
Pressure
Glass
Painted Drywall
Average Recovery (CFU)
Average Recovery (CFU)
Field
Blank
Negative
Control
Procedural
Blank
Inoculum
Control
Blank
Field
Blank
Negative
Control
Procedural
Blank
Inoculum
Control
Blank
Hype-Wipe®
Bleach Towelette
Upper left
corner (a)
Slight
ND
ND
103
ND
ND
30
ND
ND
Center (e)
Slight
ND
ND
3680
ND
ND
ND
ND
ND
Lower right
corner (i)
Slight
ND
ND
270
ND
ND
ND
ND
ND
Heavy
ND
ND
450
ND
ND
1980
1580
ND
Broad Area
Slight
ND
ND
ND
ND
ND
3870
99
ND
Heavy
ND
4
ND
ND
ND
ND
ND
ND
Clorox
Healthcare®
Bleach Germicidal
Wipe
Upper left
corner (a)
Slight
ND
500
ND
ND
ND
6
ND
ND
Center (e)
Slight
ND
ND
<200*
ND
ND
6
293
ND
Lower right
corner (i)
Slight
ND
ND
30
ND
ND
ND
ND
ND
Heavy
ND
ND
ND
ND
ND
ND
ND
ND
Broad Area
Slight
ND
524
200
ND
ND
ND
ND
ND
Heavy
ND
4
ND
ND
ND
ND
2
ND
ND: Non Detect
*: No spores detected during dilution plating; filter plating results considered outlier. Upper limit based on enumeration
detection limit of 1 spore in 0.1 mL dilution extract.
43
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5.3.2 Sample Hold Time
Due to the need for extended holding times of the samples before primary analysis (as discussed in Section
3.6.11), a testing sequence was conducted to evaluate the potential reduction of bacterial spores in the wipe
sample containers as a function of time.
The hypochlorite sporicidal wipes listed below were tested on medium size coupon surfaces to estimate the
occurrence and potential reduction of bacterial spores as a function of remediation activities. The surface
tested was glass (which represented nonporous indoor building materials, and coupons were 12.25 ft2 in
area (3.5 ft x 3.5 ft)).
a. Clorox Healthcare® Bleach Germicidal Wipe
b. Hype-Wipe® Bleach Towelette
c. pAB wipe prepared in house by soaking Fisherbrand™ dry wipes in 4 mL pAB.
The testing approach consisted of the following sequentially conducted steps:
a. The test coupons were first subjected to a specific decontamination wipe; note that the coupons were
not inoculated prior to decontamination.
b. After an exposure time of 30 minutes, the coupons were wipe sampled as follows:
areas a, b, and c were sampled together and labeled as the Day 1 sample;
areas d, e, and f were sampled and labeled as the Day 4 sample, and
Lastly, areas g, h, and i were labeled as the Day 7 sample.
c. Each of the sampled wipe sample containers was inoculated on Day 1 (one day following sampling)
with 0.1 mL of 2 x 108 CFU/mL microbial suspension (test organism solution) to result in a final
suspension that contains approximately 2 x 107 CFU of the microorganism.
d. Analysis of the samples occurred by adding 20 mL of PBST to each container on the scheduled day
(Day 1, Day 4 or Day 7), followed immediately by extracting and plating, as shown in Table 5-4.
44
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Table 5-4. Test Matrix (Test Samples)
Test ID
Decontaminant
Coupon Section
Analyzed and Plated
(Day)
Number of Wipe
Samples
73-(C/H/P)-0-G-1 -Ta-1
a
C Test Samples (3)
73-(C/H/P)-0-G-1 -Tb-1
b
Day 1
H Test Samples (3)
73-(C/H/P)-0-G-1 -T c-1
C = Clorox Healthcare®
Bleach Germicidal Wipe
c
P Test Samples (3)
73-(C/H/P)-1 -G-1 -Td-1
d
C Test Samples (3)
73-(C/H/P)-1 -G-1 -Te-1
H = Hype-Wipe® Bleach
Towelette
e
Day 4
H Test Samples (3)
73-(C/H/P)-1 -G-1 -Tf-1
P = pAB Wipe
f
P Test Samples (3)
73-(C/H/P)-7-G-1 -T g-1
g
C Test Samples (3)
73-(C/H/P)-7-G-1 -Th-1
h
Day 7
H Test Samples (3)
73-(C/H/P)-7-G-1 -Ti-1
i
P Test Samples (3)
The positive control and negative control coupons were not subjected to the decontamination wipe. The
negative control coupon and each of the positive control coupon sections were directly wipe-sampled with a
PRB wipe. While the positive coupons were inoculated with spores, the negative control coupons were not.
The procedural blank coupon was decontaminated with the hypochlorite wipe and then sampled, but did not
undergo inoculation. The control sample test matrix for this hold time study is shown in Table 5-5.
45
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Table 5-5. Test Matrix - Hold Time Control Samples
Test ID
Type
Decontaminant
Coupon
Section
Analyzed and
Plated (Day)
Number of Wipe
Samples
73-(C/H/P)-0-G-1 -Pa-1
a
C Test Samples (3)
73-(C/H/P)-0-G-1 -Pb-1
b
Day 1
H Test Samples (3)
73-(C/H/P)-0-G-1 -Pc-1
c
P Test Samples (3)
73-(C/H/P)-1 -G-1 -Pd-1
C = Clorox
d
C Test Samples (3)
73-(C/H/P)-1 -G-1 -Pe-1
Positive Control
Healthcare® Bleach
Germicidal Wipe
e
Day 4
H Test Samples (3)
73-(C/H/P)-1 -G-1 -Pf-1
H = Hype-Wipe®
Bleach Towelette
f
P Test Samples (3)
73-(C/H/P)-7-G-1 -Pg-1
P = pH-Amended
Bleach Wipe
g
C Test Samples (3)
73-(C/H/P)-7-G-1 -Ph-1
h
Day 7
H Test Samples (3)
73-(C/H/P)-7-G-1 -Pi-1
i
P Test Samples (3)
73-(C/H/P)-(0/1/7)-G-1 -XT-1
Procedural Blank
NA
0 = Day 1
1 = Day 4
7 = Day 7
1 per decontamination
wipe per day (9)
73-(C/H/P)-(0/1/7)-G-1 -NT-1
Negative Control
NA
1 per decontamination
wipe per day (9)
The results from the hold tests, summarized in Table 5-6, indicate that the samples were not affected by the
longer hold time prior to analysis. There was no reduction of bacterial spores in the sample containers as a
function of time.
46
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Table 5-6. Sample Hold Time Test Results for Decontamination Efficacy Tests
Decontamination
Wipe
Positive Controls Averaqe Recovery
(CFU)
Test Samples
Average Recovery (CFU)
LR
Day
Average
StDev
Average
StDev
Average
StDev
1
1.42 x 107
9.33 x 105
1.34 x 107
9.34 x10s
0.02
0.03
Clorox Healthcare®
4
1.40 x 107
1.04 x 106
1.35 x107
1.83 x 106
0.08
0.06
7
1.26 x 107
4.82 x 105
1.55 x107
1.08 x 106
-0.04
0.03
1
1.37 x 107
1.48 x 106
1.46 x 107
7.57 x10s
-0.03
0.02
Hype-Wipe®
4
1.44 x 107
5.18x10s
1.40 x 107
4.54 x10s
-0.01
0.01
7
1.21 x 107
4.73 x 105
1.36 x107
3.85 x10s
0.00
0.01
1
1.42 x 107
1.34 x 106
1.47 x 107
1.77 x 10s
-0.01
0.05
pAB Wipe
4
1.35 x 107
1.03 x 106
1.38 x107
7.49 x10s
0.01
0.02
7
1.29 x 107
9.05 x10s
1.27 x 107
4.23 x10s
0.05
0.01
StDev = Standard deviation.
5.4 Instrument Calibrations
The project used established and approved operating procedures for the maintenance and calibration of all
laboratory equipment. All laboratory measuring devices used in this project were certified as having been
recently calibrated or were calibrated by the on-site EPA Metrology Laboratory at the time of use. Calibration
of instruments was done at the frequency shown in Table 5-7. Any deficiencies were noted and the
instrument adjusted and recalibrated within 24 h to meet calibration tolerances. All other equipment (e.g.,
incubators, biological safety cabinets, refrigerators) used at the time of evaluation were verified as being
certified, calibrated, or validated.
47
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Table 5-7. Instrument Calibration Frequency
Equipment
Calibration/Certification
Expected Tolerance
Thermometer
Compare to independent NIST thermometer (this is a thermometer that is
recertified annually by either NIST or an International Organization for
Standardization (ISO)-17025 facility) value once per quarter
±1 °C
RH Sensor
Compare to calibration salts once a week
±5%
Stopwatch
Compare against NIST Official U.S. time at
http://nist.time.gOv/timezone.cgi7Eastern/d/-5/java once every 30 days
±1 min/30 days
Clock
Compare to office U.S. Time (® time.aov everv 30 davs
± 1 min/30 days
Scale
Check calibration with Class 2 weights
±0.1% weight
Pipettes
Certified as calibrated at time of use/recalibrated by gravimetric evaluation of
pipette performance to manufacturer's specifications every year.
±5%
pH meter
Perform a two-point calibration with standard buffers that bracket the target pH
before each use.
±0.1 pH unit
HACH High Range
Bleach Test Kit
Titrate standard solution of 1000 ppm NaCI02 before each use
± 10%
NIST: National Institute of Standards and Technology
ISO: International Organization for Standardization
5.5 QA Assessments and Response Actions
QA assessments are an integral part of a quality system. This project was assigned an EPA QA Category III
rating that merited technical system and performance audits. At regular intervals, the test team leader and
the team QA officer internally evaluated QA performance and reported the audit results to EPA management
and key project team individuals. Any identified deficiencies and corrective actions to be taken were reported
via an interoffice memorandum submitted to the responsible project participants.
An integral part of any QA program is well-defined procedures for correcting data quality problems. The
overall goals of the QA program address the following aspects of data quality:
• Problem prevention
• Problem definition
• Problem correction.
For this type of testing, data-quality problems usually require immediate, on-the-spot corrective action.
The QA assessment and action procedures followed in this project were intended to provide for rapid
detection of data quality problems. Project personnel were intimately involved with the data on a daily basis
so that any data quality issue became apparent soon after it occurred. Corrective actions were taken as soon
48
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as practical when and if a problem was observed. The nature of the problem and corrective steps taken were
noted in the project notebook of record.
5.6 Data Reduction
Data reduction for all tests performed included the total CFU recovered from each replicate coupon, the
average recovered CFU and standard deviation for each group of coupons, and LRs. For each combination
of test coupon material and sample type, the groups of coupons included the following:
• Positive control areas (replicates, average, standard deviation)
• Test areas (replicates, average, standard deviation)
• Procedural blank coupons.
Efficacy was defined as the extent (by LR) to which the agent extracted from the coupons after the treatment
with the decontamination procedure was reduced below that extracted from positive control areas (not
exposed to the decontamination procedure). The detection limit of a sample depended on the analysis
method and could therefore vary. The detection limit of a plate was assigned a value of 1 CFU, but the
fraction of the sample plated varied. For instance, the detection limit of a 0.1 mL plating of a 20 mL sample
suspension was 200 CFU (1 CFU/0.1 mL * 20 mL), but if all 20 mL of the sample were filter plated, the
detection limit was 1 CFU.
5.7 Data Reporting
Data generated included notes recorded in a laboratory notebook (e.g., gravimetric records and assessment
of decontamination solutions) and electronic files created by digital camera. Written records included
observations, numerical data produced by any instrument that was not digitally recorded, and all variables
specific to any experiment. Photographs were taken of each procedure and protocol conducted in general
and of any unusual result. Digital files were maintained in their raw form on each of two computers in the
laboratory, on desk computers used by test personnel, and on the EPA local network for backup. Processed
data files were kept on desk computers and backed up on the EPA network on a biweekly basis. Two
laboratory notebooks at a time were maintained for this project, one in the laboratory for notes related to the
inoculation and sampling procedures, and another in the NHSRC Biolab for all notes related to biological
sample analysis and coupon sterilization documentation.
5.8 QAPP Amendments and Deviations
5.8.1 Formal Amendments
During the course of this research effort, two amendments were added to the initial QAPP entitled
"Decontamination Solution Methods for Bacillus anthracis Surrogates. Sporicidal Wipes Product Assessment
Part I.", August 11, 2014. This QAPP described the Phase I testing. A first amendment described the Phase
II evaluation of the sporicidal efficacy of wipes and was approved on December 3, 2014. A second
amendment described the holding tests (see Section 5.3.2). The second amendment was provided without a
formal approval requirement.
49
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5.8.2 QAPP Deviations
One deviation was documented. The hold time test (Section 5.3.2; Amendment II to the QAPP) included pAB
wipes. The QAPP amendment text suggested that these pAB wipes were also evaluated as part of this
study. This was not the case; only four sporicidal products were evaluated in Phase I and two sporicidal
products in Phase II. There is no impact to the conclusions of this study.
50
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6 SUMMARY
Four commercially-available sporicidal wipes (Clorox Healthcare® Bleach Germicidal Wipe, Sani-Cloth®
Bleach Germicidal Disposable Wipe, Dispatch® Hospital Cleaner Disinfectant Towel with Bleach, and Hype-
Wipe® Bleach Towelettes) were evaluated on their use to decontaminate four types of materials (painted
drywall, vinyl tile, melamine board, and glass).
A first assessment involved the measurement of the amount of liquid dispensed to a surface. The individually
packaged, single-use, Hype-Wipe® Bleach Towelettes resulted in the highest volume of sporicidal solution
(bleach) dispensed to the surface of all types of materials tested. This result is not surprising as the initial
amount of liquid held by these wipes is also the highest of the four wipe materials tested. Other wipes that
are kept in bulk in their canister dispensed less liquid. Of those, the Clorox Healthcare® Wipe dispensed the
highest amount of liquid. Among the four materials tested, the least amount of liquid bleach was transferred
to glass, while painted drywall had the propensity to retain the most.
The liquid solutions dispensed from Hype-Wipe® Bleach Towelettes, and to a lesser extent from Clorox
Healthcare® Bleach Germicidal Wipes, were less susceptible to loss through evaporation from or adsorption
into the material surface over time. All decontamination wipes dispensed enough liquid (with bleach as the
active ingredient) to keep the tested surfaces "wet" based on visual observation over the five-min contact time.
A higher pressure application resulted in higher liquid solution volume dispensed compared to slight (low)
pressure application for all material/wipe type combinations. For these particular tests, the effect of applying
higher pressure resulted in an increase of approximately 60% in liquid being dispensed from the Hype-Wipe®
Bleach Towelettes to the coupon and up to a 300% increase in liquid from the Sani-Cloth® wipes.
Two sporicidal wipes were selected for further evaluation of their efficacy to decontaminate medium size
surfaces. The decontamination efficacies of the Hype-Wipe® Bleach Towelette and Clorox Healthcare® Wipe
were evaluated on medium-size surface areas (12.25 ft2,1.37 m2) for different inoculation methods (Hot Spot
versus Broad Area) and wipe application pressure method (slight versus high pressure).
Results indicate that the Hype-Wipe® Bleach Towelette was slightly more effective in inactivating Bg spores
than the Clorox Healthcare® Wipe. This difference may be attributed to the higher amount of liquid that was
dispensed from the Hype-Wipe® Bleach Towelette when compared to the Clorox Healthcare® Wipe. The
overall log reduction in viable spores, including all inoculation conditions and both materials, by the Hype-
Wipe® Bleach Towelette and Clorox Healthcare® Wipe was 3.8 ± 0.8 and 3.0 ± 1.0, respectively. The wipe
application pressure seems to have little or no effect on the decontamination efficacy of the wipe. Three
sporicidal wipe tests may have been biased by significant number of spores (>500) on procedural blank or
negative control coupons. Excluding results from those tests does not change the average log reduction in
viable spores across all inoculation conditions for either sporicidal wipe.
The decontamination wipes that were used during the decontamination process were stored for a minimum
of 24 h at 4 °C in sterile specimen cups. No viable Bg spores were detected on these wipes after use.
Post-decontamination sampling of coupons showed that cross contamination occurs, especially when the
exposed surface area of the bleach decontamination wipe is drying out due to the wiping of dry surface
51
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areas. This cross contamination seems to have started with the contamination of the spent surface of the
wipe, when it dried, resulting in contamination of other areas of the coupons during subsequent wiping.
Neither sporicidal wipe meets the 6 log reduction in viable spores when wiping a larger 12.25 ft2 (1.37 m2)
surface area. The presence of a significant number of viable spores on some of the procedural blanks or
negative controls may bias this statement negatively. However, the overall impact of this cross
contamination should be considered minimal as the vast majority of tests resulted in a less than 6 log
reduction in viable spores. These results are in contrast to the previously reported results where >6 log
reductions were achieved when wiping a 1 ft2 surface area [1], Further testing should be considered to find
the maximum area these wipes can be used by conducting efficacy tests to verify a 6 LR.
52
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7 REFERENCES
1. K.M. Meyer, J.A.Tufts., M.W. Calfee, and L. Oudejans, Efficacy of sporicidal wipes for inactivation of
a Bacillus anthracis surrogate. Journal of Applied Microbiology, 2014.117(6): 1634-44.
2 C.P. Weis, A.J. Intrepido, A.K. Miller, P.G. Cowin, M.A. Durno, J.S. Gebhart, and R. Bull, Secondary
aerosolization of viable Bacillus anthracis spores in a contaminated U.S. Senate office, Journal of the
American Medical Association, 2002. 288(22): 2853-8.
3. U.S. EPA, Office of Research and Development, Systematic Investigation of Liquid and Fumigant
Decontamination Efficacy against Biological Agents Deposited on Test Coupons of Common Indoor
Materials. U.S. Environmental Protection Agency, Washington, DC, EPA/600/R-11/076, 2011.
4. G.S. Brown, R.G. Betty, J.E. Brockmann, D.A. Lucero, C.A. Souza, K.S. Walsh, R.M. Boucher, M.
Tezak, M.C. Wilson, and T. Rudolph, Evaluation of a wipe surface sample method for collection of
Bacillus spores from nonporous surfaces. Applied and Environmental Microbiology, 2007. 73(3):
706-710.
5. M.W. Calfee, S.D. Lee, and S.P. Ryan, A rapid and repeatable method to deposit bioaerosols on
material surfaces. Journal of Microbiological Methods, 2013. 92(3): 375-380.
6. S.D. Lee, S.P. Ryan, and E.G. Snyder, Development of an aerosol surface inoculation method for
Bacillus spores. Applied and Environmental Microbiology, 2011. 77(5): 1638-1645.
7. M.W. Calfee, S.P. Ryan, J.P. Wood, L. Mickelsen, C. Kempter, L. Miller, M. Colby, A. Touati, M.
Clayton, N. Griffin-Gatchalian, S. McDonald and R. Delafield, Laboratory evaluation of large-scale
decontamination approaches. Journal of Applied Microbiology, 2012.112(5): 874-82.
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APPENDIX A - PHASE 1: Operational Aspects of Decontamination by Sporicidal
Wipes
Test Results for Task 1: Determination of Effective Surface Coverage of Sporicidal Wipes
Table A-1-1-1: Task 1 Results Clorox Healthcare® Bleach Germicidal Wipe/Vinyl Flooring
Wipe
Size
ID
Random
Weight of Wet
Net
Net Wipe
Net Weight
Average
StDev
Type
Number
Assignment
Initial
Final
Weight
Loss (g)
Time
(seconds)
Loss (g/ft2)
Weight
Loss (g/ft2)
Weight
Loss (g/ft2)
WA73-1-V-1-W1-1
7
13.58
11.25
2.33
19.00
2.33
WA73-1 -V-1 -W1 -2
24
12.91
10.54
2.37
18.00
2.37
1x1
WA73-1 -V-1 -W1 -3
5
13.23
10.88
2.36
19.00
2.36
2.39
0.06
WA73-1 -V-1 -W1 -4
19
12.46
9.99
2.47
17.00
2.47
WA73-1 -V-1 -W1 -5
10
12.55
10.13
2.42
18.00
2.42
WA73-1-V-1-W2-1
4
12.44
8.98
3.47
25.00
1.54
CT>
C
WA73-1 -V-1 -W2-2
12
12.34
8.94
3.40
24.00
1.51
o
_o
u_
1.5x1.5
WA73-1 -V-1 -W2-3
23
13.04
9.65
3.39
24.00
1.51
1.50
0.04
¦>¦>
c
WA73-1 -V-1 -W2-4
11
11.98
8.74
3.24
28.00
1.44
s
0)
Q_
WA73-1 -V-1 -W2-5
9
12.57
9.15
3.42
24.00
1.52
§
CT3
WA73-1-V-1-W3-1
1
12.91
8.89
4.03
34.00
1.01
TD
'o
'£
k_
0)
O
WA73-1 -V-1 -W3-2
21
12.28
8.21
4.06
34.00
1.02
2x2
WA73-1 -V-1 -W3-3
22
12.98
8.73
4.24
36.00
1.06
1.01
0.03
.=
o
tc
0)
CD
WA73-1 -V-1 -W3-4
8
12.54
8.62
3.92
33.00
0.98
WA73-1 -V-1 -W3-5
15
13.11
9.07
4.05
34.00
1.01
k_
tc
o
WA73-1-V-1-W4-1
6
14.34
9.55
4.79
42.00
0.77
tc
0)
WA73-1 -V-1 -W4-2
13
12.55
8.07
4.48
44.00
0.72
X
X
o
2.5x2.5
WA73-1 -V-1 -W4-3
14
12.06
7.65
4.42
43.00
0.71
0.75
0.04
o
o
WA73-1 -V-1 -W4-4
25
13.56
8.82
4.74
43.00
0.76
WA73-1 -V-1 -W4-5
3
12.66
7.58
5.07
42.00
0.81
WA73-1-V-1-W5-1
18
12.59
7.33
5.26
51.00
0.58
WA73-1 -V-1 -W5-2
16
13.05
7.82
5.23
53.00
0.58
3x3
WA73-1 -V-1 -W5-3
20
12.53
7.26
5.27
54.00
0.59
0.58
0.01
WA73-1 -V-1 -W5-4
2
13.23
7.87
5.36
52.00
0.60
WA73-1 -V-1 -W5-5
17
12.27
7.09
5.18
50.00
0.58
54
-------
Table A-1-1-2: Task 1 Results Clorox Healthcare® Bleach Germicidal Wipe/Glass
Wipe
Type
Random
Weight of Wet
Net
Net Wipe
Net
Average
StDev
Size
ID
Number
Wipes (g)
Weight
Time
Weight
Weight
Weight
Assignment
Initial
Final
Loss
(seconds)
Loss
Loss (g/ft2)
Loss (g/ft2)
fcrt
fa/ft2l
WA73-1-G-1-W1-1
13
12.17
10.72
1.45
21.00
1.45
WA73-1-G-1-W1-2
11
12.08
10.61
1.46
20.00
1.46
1x1
WA73-1-G-1-W1-3
6
11.96
10.68
1.29
21.00
1.29
1.36
0.10
WA73-1-G-1-W14
10
12.43
11.06
1.37
18.00
1.37
WA73-1-G-1-W1-5
5
12.89
11.67
1.22
19.00
1.22
WA73-1-G-1-W2-1
20
12.33
9.71
2.62
22.00
1.16
WA73-1-G-1-W2-2
21
12.63
9.99
2.64
24.00
1.17
cn
O
WA73-1-G-1-W3-1
15
12.13
8.90
3.22
28.00
0.81
WA73-1-G-1-W3-2
22
12.25
9.02
3.23
26.00
0.81
.=
o
tc
a>
CD
2x2
WA73-1-G-1-W3-3
16
12.57
9.12
3.45
29.00
0.86
0.83
0.02
WA73-1-G-1-W34
23
12.22
8.86
3.36
31.00
0.84
k_
tc
o
WA73-1-G-1-W3-5
4
12.10
8.84
3.26
32.00
0.82
tc
a>
WA73-1-G-1-W4-1
19
12.36
8.43
3.92
40.00
0.63
X
X
s
WA73-1-G-1-W4-2
7
12.30
8.17
4.13
39.00
0.66
o
o
2.5x2.5
WA73-1-G-1-W4-3
25
12.28
8.25
4.03
37.00
0.64
0.65
0.02
WA73-1-G-1-W44
2
12.50
8.33
4.17
38.00
0.67
WA73-1-G-1-W4-5
24
12.42
8.27
4.15
35.00
0.66
WA73-1-G-1-W5-1
17
12.19
7.56
4.63
56.00
0.51
WA73-1-G-1-W5-2
1
13.09
8.61
4.48
55.00
0.50
3x3
WA73-1-G-1-W5-3
9
11.82
7.34
4.47
51.00
0.50
0.52
0.03
WA73-1-G-1-W54
8
12.63
7.99
4.64
53.00
0.52
WA73-1-G-1-W5-5
17
12.27
7.09
5.18
54.00
0.58
55
-------
Table A-1-1-3: Task 1 Results Clorox Healthcare® Bleach Germicidal Wipe/Melamine
Wipe
Type
Size
ID
Random
Number
Assignment
Weight of Wet
Wipes (g)
Net
Weight
Loss (g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss (g/ft2)
Initial
Final
Clorox Healthcare® Bleach Germicidal Wipe/Melamine
1x1
WA73-1-M-1-W1-1
3
12.10
10.88
1.22
18.00
1.22
1.25
0.07
WA73-1-M-1-W1-2
25
11.92
10.55
1.38
19.00
1.38
WA73-1-M-1-W1-3
23
11.46
10.23
1.22
20.00
1.22
WA73-1-M-1-W1-4
21
11.78
10.59
1.20
18.00
1.20
WA73-1-M-1-W1-5
7
11.63
10.39
1.24
20.00
1.24
1.5x1.5
WA73-1-M-1-W2-1
6
12.65
9.84
2.81
25.00
1.25
1.20
0.05
WA73-1-M-1-W2-2
2
12.19
9.37
2.82
27.00
1.26
WA73-1-M-1-W2-3
24
11.62
9.04
2.58
24.00
1.15
WA73-1-M-1-W2-4
15
12.28
9.59
2.68
26.00
1.19
WA73-1-M-1-W2-5
12
12.73
10.11
2.63
27.00
1.17
2x2
WA73-1-M-1-W3-1
9
11.60
8.34
3.26
36.00
0.82
0.83
0.02
WA73-1-M-1-W3-2
4
12.07
8.80
3.27
37.00
0.82
WA73-1-M-1-W3-3
8
12.12
8.81
3.32
35.00
0.83
WA73-1-M-1-W3-4
20
13.09
9.84
3.25
35.00
0.81
WA73-1-M-1-W3-5
16
12.20
8.73
3.47
34.00
0.87
2.5x2.5
WA73-1-M-1-W4-1
18
12.82
8.16
4.66
44.00
0.75
0.75
0.01
WA73-1-M-1-W4-2
14
11.25
6.48
4.77
45.00
0.76
WA73-1-M-1-W4-3
5
11.88
7.29
4.58
48.00
0.73
WA73-1-M-1-W4-4
19
13.00
8.33
4.68
51.00
0.75
WA73-1-M-1-W4-5
11
11.99
7.36
4.63
48.00
0.74
3x3
WA73-1-M-1-W5-1
10
12.16
6.98
5.18
56.00
0.58
0.57
0.01
WA73-1-M-1-W5-2
17
12.25
7.23
5.02
56.00
0.56
WA73-1-M-1-W5-3
22
13.25
8.00
5.26
53.00
0.58
WA73-1-M-1-W5-4
13
12.03
6.89
5.14
52.00
0.57
WA73-1-M-1-W5-5
17
12.27
7.09
5.18
55.00
0.58
56
-------
Table A-1-1-4: Task 1 Results Clorox Healthcare® Bleach Germicidal Wipe/Painted Drywall
Wipe
Type
Size
ID
Random
Number
Assignment
Weight
Wipe
Initial
of Wet
s(g)
Final
Net
Weight
Loss (g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
g/(ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss (g/ft2)
Clorox Healthcare® Bleach Germicidal Wipe/Painted Drywall
1x1
WA73-1-D-1-W1-1
16
12.21
10.44
1.77
27.00
1.77
1.74
0.02
WA73-1-D-1-W1-2
9
11.07
9.36
1.71
25.00
1.71
WA73-1-D-1-W1-3
24*
11.48
9.75
1.73
21.00
1.73
WA73-1-D-1-W1-4
2
12.15
10.40
1.75
24.00
1.75
WA73-1-D-1-W1-5
17
11.52
9.80
1.72
23.00
1.72
1.5x1.5
WA73-1-D-1-W2-1
4
11.44
8.37
3.07
34.00
1.37
1.34
0.04
WA73-1-D-1-W2-2
7
12.00
9.01
2.99
33.00
1.33
WA73-1-D-1-W2-3
12
11.48
8.58
2.89
35.00
1.29
WA73-1-D-1-W2-4
21
12.11
9.10
3.01
33.00
1.34
WA73-1-D-1-W2-5
14
12.27
9.13
3.14
34.00
1.39
2x2
WA73-1-D-1-W3-1
15
12.09
8.11
3.98
41.00
0.99
1.00
0.01
WA73-1-D-1-W3-2
13
12.12
8.07
4.05
46.00
1.01
WA73-1-D-1-W3-3
1
12.10
8.03
4.07
44.00
1.02
WA73-1-D-1-W3-4
22
12.15
8.18
3.97
41.00
0.99
WA73-1-D-1-W3-5
19
12.33
8.37
3.96
46.00
0.99
2.5x2.5
WA73-1-D-1-W4-1
25*
12.25
7.11
5.14
48.00
0.82
0.83
0.01
WA73-1-D-1-W4-2
6
12.10
6.82
5.29
51.00
0.85
WA73-1-D-1-W4-3
8
12.18
7.08
5.10
54.00
0.82
WA73-1-D-1-W4-4
3
11.81
6.68
5.13
52.00
0.82
WA73-1-D-1-W4-5
10
12.21
6.99
5.22
51.00
0.83
3x3
WA73-1-D-1-W5-1
5
12.22
6.10
6.11
62.00
0.68
0.66
0.05
WA73-1-D-1-W5-2
20
11.50
5.49
6.01
62.00
0.67
WA73-1-D-1-W5-3
18
12.21
5.93
6.28
67.00
0.70
WA73-1-D-1-W5-4
23
11.52
5.42
6.11
61.00
0.68
WA73-1-D-1-W5-5
17
12.27
7.09
5.18
66.00
0.58
* 25 completed before 24
57
-------
Table A-1-2-1: Task 1 Results Hype-Wipe® Bleach Towelette/Vinyl Flooring
Wipe
Type
Size
ID
Random
Number
Assignment
Weight
Wipe
Initial
of Wet
s(g)
Final
Net
Weight
Loss (g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
g/(ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss
(g/ft2)
Hype-Wipe® Bleach Towelette/Vinyl Flooring
1x1
WA73-1-V-4-W1-1
6
15.02
11.83
3.19
28.00
3.19
3.16
0.66
WA73-1-V4-W1 -2
22
14.07
12.01
2.05
33.00
2.05
WA73-1-V4-W1 -3
13
15.20
11.51
3.69
23.00
3.69
WA73-1-V4-W1 -4
20
15.11
11.86
3.25
24.00
3.25
WA73-1-V4-W1 -5
12
15.11
11.46
3.64
24.00
3.64
1.5x1.5
WA73-1-V4-W2-1
14
15.14
10.63
4.51
38.00
2.00
2.01
0.06
WA73-1-V4-W2-2
23
15.06
10.70
4.36
35.00
1.94
WA73-1-V4-W2-3
8
15.07
10.60
4.46
41.00
1.98
WA73-1-V4-W2-4
2
15.08
10.37
4.71
45.00
2.09
WA73-1-V4-W2-5
4
15.01
10.47
4.54
49.00
2.02
2x2
WA73-1-V-4-W3-1
16
15.09
9.32
5.77
44.00
1.44
1.37
0.05
WA73-1-V4-W3-2
1
15.01
9.46
5.55
53.00
1.39
WA73-1-V4-W3-3
25
15.00
9.55
5.45
47.00
1.36
WA73-1-V4-W3-4
3
14.93
9.73
5.20
55.00
1.30
WA73-1-V4-W3-5
24
15.01
9.51
5.50
50.00
1.37
2.5x2.5
WA73-1-V4-W4-1
15
15.13
8.85
6.28
62.00
1.01
1.01
0.02
WA73-1-V4-W4-2
11
15.00
8.65
6.35
57.00
1.02
WA73-1-V4-W4-3
18
15.24
8.74
6.50
57.00
1.04
WA73-1-V4-W4-4
10
15.09
8.76
6.34
57.00
1.01
WA73-1-V4-W4-5
9
15.03
8.84
6.19
58.00
0.99
3x3
WA73-1-V-4-W5-1
7
15.05
8.85
6.20
75.00
0.69
0.71
0.09
WA73-1-V4-W5-2
21
15.07
8.39
6.68
72.00
0.74
WA73-1-V4-W5-3
19
15.02
8.03
7.00
76.00
0.78
WA73-1-V4-W5-4
17
15.08
8.04
7.04
73.00
0.78
WA73-1-V4-W5-5
17
12.27
7.09
5.18
78.00
0.58
58
-------
Table A-1-2-2: Task 1 Results Hype-Wipe® Bleach Towelette/Glass
Wipe
Type
Size
ID
Random
Number
Assignment
Weight
Wipe
Initial
of Wet
5(9)
Final
Net
Weight
Loss
(g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss
(g/ft2)
Hype-Wipe® Bleach Towelette/Glass
1x1
WA73-1-G-4-W1-1
19
14.97
12.55
2.42
24.00
2.42
2.51
0.17
WA73-1-G-4-W1-2
20
14.99
12.67
2.32
22.00
2.32
WA73-1-G-4-W1-3
4
15.38
12.87
2.51
28.00
2.51
WA73-1-G-4-W1-4
16
14.97
12.42
2.55
27.00
2.55
WA73-1-G-4-W1-5
1
15.01
12.25
2.76
25.00
2.76
1.5x1.5
WA73-1-G-4-W2-1
7
15.06
11.28
3.78
31.00
1.68
1.77
0.09
WA73-1-G-4-W2-2
21
15.03
11.09
3.94
33.00
1.75
WA73-1-G-4-W2-3
13
15.18
10.98
4.21
34.00
1.87
WA73-1-G-4-W2-4
9
15.04
11.20
3.84
32.00
1.71
WA73-1-G-4-W2-5
25
15.25
11.06
4.19
32.00
1.86
2x2
WA73-1-G-4-W3-1
6
15.28
10.16
5.13
41.00
1.28
1.23
0.03
WA73-1-G-4-W3-2
8
15.15
10.22
4.94
40.00
1.23
WA73-1-G-4-W3-3
22
14.95
10.08
4.86
38.00
1.22
WA73-1-G-4-W3-4
23
15.24
10.29
4.95
38.00
1.24
WA73-1-G-4-W3-5
15
14.56
9.76
4.80
39.00
1.20
2.5x2.5
WA73-1-G-4-W4-1
10
15.15
9.72
5.43
49.00
0.87
0.85
0.02
WA73-1-G-4-W4-2
2
15.00
9.61
5.39
57.00
0.86
WA73-1-G-4-W4-3
3
15.06
9.85
5.22
50.00
0.83
WA73-1-G-4-W4-4
5
15.09
9.73
5.36
52.00
0.86
WA73-1-G-4-W4-5
18
15.11
9.98
5.14
49.00
0.82
3x3
WA73-1-G-4-W5-1
14
14.94
8.95
5.99
66.00
0.67
0.63
0.04
WA73-1-G-4-W5-2
11
15.00
9.47
5.53
63.00
0.61
WA73-1-G-4-W5-3
12
15.09
9.31
5.79
64.00
0.64
WA73-1-G-4-W5-4
24
15.16
9.25
5.91
63.00
0.66
WA73-1-G-4-W5-5
17
12.27
7.09
5.18
62.00
0.58
59
-------
Table A-1-2-3: Task 1 Results Hype-Wipe® Bleach Towelette/Melamine
Wipe
Type
Size
ID
Random
Number
Assignment
Weight of Wet
Wipes (g)
Net
Weight
Loss (g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss (g/ft2)
Initial
Final
WA73-1-M-4-W1-1
17
15.02
11.55
3.47
24.00
3.47
WA73-1-M-4-W1-2
23
14.94
11.57
3.37
27.00
3.37
1x1
WA73-1-M-4-W1-3
12
14.97
11.51
3.46
31.00
3.46
3.62
0.31
WA73-1-M-4-W1-4
5
14.99
11.31
3.68
28.00
3.68
WA73-1-M-4-W1-5
11
15.00
10.87
4.14
30.00
4.14
WA73-1-M-4-W2-1
3
15.01
9.77
5.25
40.00
2.33
WA73-1-M-4-W2-2
2
15.09
9.69
5.40
48.00
2.40
0>
c
E
1.5x1.5
WA73-1-M-4-W2-3
10
15.18
10.44
4.74
42.00
2.11
2.29
0.12
WA73-1-M-4-W2-4
4
15.10
9.76
5.34
46.00
2.37
CO
0>
WA73-1-M-4-W2-5
15
15.22
10.24
4.98
47.00
2.21
"a>
o>
0>
o
I—
JZ.
WA73-1-M-4-W3-1
1
15.07
9.70
5.37
48.00
1.34
WA73-1-M-4-W3-2
22
14.98
9.52
5.45
62.00
1.36
2x2
WA73-1-M-4-W3-3
9
15.17
9.78
5.39
56.00
1.35
1.37
0.03
fZ
0>
QQ
©
0)
Q_
&
Q.
>>
X
WA73-1-M-4-W3-4
19
14.86
9.27
5.59
60.00
1.40
WA73-1-M-4-W3-5
20
15.07
9.48
5.58
63.00
1.40
WA73-1-M-4-W4-1
16
15.25
9.06
6.20
63.00
0.99
WA73-1-M-4-W4-2
24
15.17
9.13
6.04
62.00
0.97
2.5x2.5
WA73-1-M-4-W4-3
7
15.22
8.67
6.55
66.00
1.05
1.02
0.04
WA73-1-M-4-W4-4
18
15.06
8.60
6.46
68.00
1.03
WA73-1-M-4-W4-5
6
15.13
8.64
6.49
65.00
1.04
WA73-1-M-4-W5-1
14
14.95
8.09
6.87
86.00
0.76
WA73-1-M-4-W5-2
8
15.13
8.04
7.09
81.00
0.79
3x3
WA73-1-M-4-W5-3
21
15.08
7.99
7.09
83.00
0.79
0.73
0.09
WA73-1-M-4-W5-4
25
14.97
8.17
6.80
81.00
0.76
WA73-1-M-4-W5-5
17
12.27
7.09
5.18
82.00
0.58
60
-------
Table A-1-2-4: Task 1 Results Hype-Wipe® Bleach Towelette/Painted Drywall
Wipe
Type
Size
ID
Random
Number
Assignment
Weight of Wet
Wipes (g)
Net
Weight
Loss
(g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss
(g/ft2)
Initial
Final
Hype-Wipe® Bleach Towelette/Painted Drywall
1x1
WA73-1-D-4-W1-1
11
15.03
11.83
3.20
27.00
3.20
3.35
0.33
WA73-1-D-4-W1-2
18
15.01
11.73
3.28
30.00
3.28
WA73-1-D-4-W1-3
12
14.93
11.20
3.73
32.00
3.73
WA73-1-D-4-W1-4
2
15.12
12.20
2.92
28.00
2.92
WA73-1-D-4-W1-5
15
14.89
11.25
3.64
28.00
3.64
1.5x1.5
WA73-1-D-4-W2-1
21
14.91
10.10
4.81
37.00
2.14
2.20
0.11
WA73-1-D-4-W2-2
20
14.92
9.67
5.25
68.00
2.34
WA73-1-D-4-W2-3
25
15.02
10.10
4.92
39.00
2.19
WA73-1-D-4-W2-4
17
14.79
10.13
4.67
37.00
2.08
WA73-1-D-4-W2-5
5
15.07
9.93
5.14
42.00
2.28
2x2
WA73-1-D-4-W3-1
16
14.96
9.51
5.45
54.00
1.36
1.42
0.04
WA73-1-D-4-W3-2
7
15.04
9.17
5.87
57.00
1.47
WA73-1-D-4-W3-3
14
14.88
9.19
5.70
44.00
1.42
WA73-1-D-4-W3-4
24
15.11
9.33
5.78
53.00
1.45
WA73-1-D-4-W3-5
6
15.07
9.45
5.62
60.00
1.40
2.5x2.5
WA73-1-D-4-W4-1
22
15.17
8.40
6.77
58.00
1.08
1.06
0.05
WA73-1-D-4-W4-2
10
14.95
8.75
6.20
63.00
0.99
WA73-1-D-4-W4-3
19
15.00
8.63
6.37
57.00
1.02
WA73-1-D-4-W4-4
3
15.09
8.11
6.99
65.00
1.12
WA73-1-D-4-W4-5
9
15.00
8.14
6.86
55.00
1.10
3x3
WA73-1-D-4-W5-1
13
15.11
7.27
7.84
89.00
0.87
0.80
0.13
WA73-1-D-4-W5-2
23
14.99
7.30
7.69
83.00
0.85
WA73-1-D-4-W5-3
8
14.92
7.23
7.69
79.00
0.85
WA73-1-D-4-W5-4
1
15.04
7.24
7.80
78.00
0.87
WA73-1-D-4-W5-5
17
12.27
7.09
5.18
79.00
0.58
61
-------
Table A-1-3-1: Task 1 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Vinyl Flooring
Wipe
Type
Size
ID
Random
Number
Assignment
Weight of Wet
Wipes (g)
Net
Weight
Loss
(g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss
(g/ft2)
Initial
Final
Sani-Cloth® Bleach Germicidal Disposable Wipe/Vinyl Flooring
1x1
WA73-1-V-2-W1-1
14
11.03
9.94
1.10
17.00
1.10
1.22
0.16
WA73-1-V-2-W1-2
15
11.10
9.92
1.18
17.00
1.18
WA73-1-V-2-W1-3
5
10.28
9.15
1.13
20.00
1.13
WA73-1-V-2-W1-4
24
10.85
9.64
1.20
21.00
1.20
WA73-1-V-2-W1-5
22
11.01
9.52
1.49
21.00
1.49
1.5x1.5
WA73-1-V-2-W2-1
2
10.59
7.68
2.92
28.00
1.30
1.25
0.05
WA73-1-V-2-W2-2
13
10.38
7.67
2.71
28.00
1.20
WA73-1-V-2-W2-3
19
10.25
7.31
2.94
29.00
1.31
WA73-1-V-2-W2-4
10
10.77
8.08
2.69
26.00
1.20
WA73-1-V-2-W2-5
20
11.20
8.40
2.79
26.00
1.24
2x2
WA73-1-V-2-W3-1
12
11.10
7.76
3.35
37.00
0.84
0.88
0.04
WA73-1-V-2-W3-2
18
11.30
7.61
3.69
39.00
0.92
WA73-1-V-2-W3-3
21
11.42
7.90
3.52
33.00
0.88
WA73-1-V-2-W3-4
6
10.47
7.02
3.45
36.00
0.86
WA73-1-V-2-W3-5
25
11.08
7.44
3.64
32.00
0.91
2.5x2.5
WA73-1-V-2-W4-1
17
10.26
5.39
4.87
66.00
0.78
0.77
0.03
WA73-1-V-2-W4-2
7
10.59
5.53
5.06
48.00
0.81
WA73-1-V-2-W4-3
8
10.22
5.64
4.58
45.00
0.73
WA73-1-V-2-W4-4
1
10.39
5.70
4.69
45.00
0.75
WA73-1-V-2-W4-5
4
10.60
5.61
5.00
43.00
0.80
3x3
WA73-1-V-2-W5-1
9
10.83
5.78
5.05
57.00
0.56
0.57
0.01
WA73-1-V-2-W5-2
11
10.41
5.25
5.16
52.00
0.57
WA73-1-V-2-W5-3
23
10.71
5.44
5.27
56.00
0.59
WA73-1-V-2-W5-4
16
10.21
5.20
5.01
56.00
0.56
WA73-1-V-2-W5-5
17
12.27
7.09
5.18
66.00
0.58
62
-------
Table A-1-3-2: Task 1 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Glass
Wipe
Type
Size
ID
Random
Number
Assignment
Weight of Wet
Wipes (g)
Net
Weight
Loss (g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss
(g/ft2)
Initial
Final
WA73-1-G-2-W1-1
10
11.12
10.20
0.92
18.00
0.92
WA73-1-G-2-W1-2
17
11.06
9.88
1.18
17.00
1.18
1x1
WA73-1-G-2-W1-3
7
10.64
9.72
0.92
19.00
0.92
0.99
0.11
WA73-1-G-2-W1-4
8
10.74
9.79
0.95
17.00
0.95
WA73-1-G-2-W1-5
19
10.67
9.68
0.99
19.00
0.99
WA73-1-G-2-W2-1
15
11.07
9.99
1.07
23.00
0.48
>
>
CO
O
WA73-1-G-2-W2-2
23
11.03
9.77
1.26
24.00
0.56
1.5x1.5
WA73-1-G-2-W2-3
3
10.82
9.81
1.02
23.00
0.45
0.51
0.05
CU
Q.
g
WA73-1-G-2-W2-4
18
10.81
9.58
1.23
25.00
0.55
CU
¦Q
CO
(/)
O
Q.
(/)
Q
"co
T3
WA73-1-G-2-W2-5
13
10.98
9.79
1.19
25.00
0.53
WA73-1-G-2-W3-1
2
10.94
8.07
2.86
32.00
0.72
WA73-1-G-2-W3-2
9
11.14
8.14
3.00
33.00
0.75
2x2
WA73-1-G-2-W3-3
16
11.06
8.34
2.71
34.00
0.68
0.72
0.03
E
L_
cu
O
¦E
o
WA73-1-G-2-W3-4
5
11.05
8.08
2.96
33.00
0.74
WA73-1-G-2-W3-5
24
10.69
7.84
2.86
33.00
0.71
cu
CD
WA73-1-G-2-W4-1
1
11.13
6.96
4.17
40.00
0.67
.=
o
O
'E
CO
WA73-1-G-2-W4-2
11
11.18
6.85
4.33
39.00
0.69
2.5x2.5
WA73-1-G-2-W4-3
22
11.16
6.77
4.40
40.00
0.70
0.69
0.01
CO
WA73-1-G-2-W4-4
21
10.83
6.48
4.35
42.00
0.70
WA73-1-G-2-W4-5
25
11.01
6.70
4.31
39.00
0.69
WA73-1-G-2-W5-1
12
10.92
6.09
4.82
51.00
0.54
WA73-1-G-2-W5-2
14
10.77
5.57
5.20
54.00
0.58
3x3
WA73-1-G-2-W5-3
20
10.92
5.64
5.28
59.00
0.59
0.57
0.02
WA73-1-G-2-W5-4
4
11.01
6.06
4.95
55.00
0.55
WA73-1-G-2-W5-5
17
12.27
7.09
5.18
55.00
0.58
63
-------
Table A-1-3-3: Task 1 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Melamine
Wipe
Type
Size
ID
Random
Number
Assignment
Weig
WetV
(g
Initial
ht of
Vipes
)
Final
Net
Weight
Loss (g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss (g/ft2)
Sani-Cloth® Bleach Germicidal Disposable Wipe/Melamine
1x1
WA73-1-M-2-W1 -1
22
10.70
9.65
1.05
18.00
1.05
1.13
0.09
WA73-1-M-2-W1 -2
2
11.00
9.82
1.18
19.00
1.18
WA73-1-M-2-W1 -3
10
11.05
9.96
1.09
20.00
1.09
WA73-1-M-2-W1 -4
3
10.81
9.75
1.06
19.00
1.06
WA73-1-M-2-W1 -5
21
10.70
9.44
1.25
19.00
1.25
1.5x1.5
WA73-1-M-2-W2-1
6
11.02
8.85
2.17
29.00
0.97
0.91
0.04
WA73-1-M-2-W2-2
11
10.83
8.70
2.13
25.00
0.94
WA73-1-M-2-W2-3
5
10.39
8.39
2.00
30.00
0.89
WA73-1-M-2-W2-4
24
10.60
8.66
1.94
27.00
0.86
WA73-1-M-2-W2-5
18
10.96
8.93
2.03
29.00
0.90
2x2
WA73-1-M-2-W3-1
15
10.61
7.31
3.30
30.00
0.82
0.83
0.03
WA73-1-M-2-W3-2
25
10.58
7.45
3.13
31.00
0.78
WA73-1-M-2-W3-3
8
11.06
7.60
3.46
35.00
0.87
WA73-1-M-2-W3-4
20
11.00
7.65
3.34
31.00
0.84
WA73-1-M-2-W3-5
9
11.01
7.72
3.29
34.00
0.82
2.5x2.5
WA73-1-M-2-W4-1
23
11.30
7.05
4.25
34.00
0.68
0.70
0.02
WA73-1-M-2-W4-2
16
10.77
6.50
4.28
45.00
0.68
WA73-1-M-2-W4-3
17
11.06
6.54
4.52
47.00
0.72
WA73-1-M-2-W4-4
14
11.10
6.57
4.53
43.00
0.72
WA73-1-M-2-W4-5
1
10.68
6.36
4.32
43.00
0.69
3x3
WA73-1-M-2-W5-1
7
10.41
5.26
5.16
58.00
0.57
0.58
0.01
WA73-1-M-2-W5-2
13
10.65
5.46
5.18
50.00
0.58
WA73-1-M-2-W5-3
19
10.65
5.50
5.14
49.00
0.57
WA73-1-M-2-W5-4
12
10.93
5.61
5.32
51.00
0.59
WA73-1-M-2-W5-5
17
12.27
7.09
5.18
52.00
0.58
64
-------
Table A-1-3-4: Task 1 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Painted Drywall
Wipe
Type
Size
ID
Random
Number
Assignment
Weight of
Wet Wipes
(g)
Net
Weight
Loss (g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss (g/ft2)
Initial
Final
Sani-Cloth8 Bleach Germicidal Disposable Wipe/Painted Drywall
1x1
WA73-1-D-2-W1-1
24
11.02
9.42
1.61
25.00
1.61
1.56
0.05
WA73-1-D-2-W1-2
23
11.27
9.66
1.61
26.00
1.61
WA73-1-D-2-W1-3
17
10.84
9.35
1.49
22.00
1.49
WA73-1-D-2-W1-4
2
10.56
9.05
1.52
22.00
1.52
WA73-1-D-2-W1-5
4
10.63
9.09
1.55
22.00
1.55
1.5x1.5
WA73-1-D-2-W2-1
1
10.64
7.92
2.71
32.00
1.21
1.31
0.07
WA73-1-D-2-W2-2
21
10.92
7.86
3.06
32.00
1.36
WA73-1-D-2-W2-3
18
11.16
8.07
3.10
35.00
1.38
WA73-1-D-2-W2-4
22
11.01
7.99
3.03
34.00
1.34
WA73-1-D-2-W2-5
5
10.95
8.09
2.86
35.00
1.27
2x2
WA73-1-D-2-W3-1
3
10.53
6.27
4.26
42.00
1.06
1.09
0.05
WA73-1-D-2-W3-2
14
11.20
7.06
4.14
NA
1.04
WA73-1-D-2-W3-3
20
11.19
6.67
4.52
43.00
1.13
WA73-1-D-2-W3-4
6
10.69
6.32
4.36
45.00
1.09
WA73-1-D-2-W3-5
11
11.22
6.64
4.58
46.00
1.15
2.5x2.5
WA73-1-D-2-W4-1
9
11.16
5.52
5.65
52.00
0.90
0.91
0.02
WA73-1-D-2-W4-2
13
10.90
5.15
5.75
53.00
0.92
WA73-1-D-2-W4-3
19
11.24
5.36
5.88
86.00
0.94
WA73-1-D-2-W4-4
7
10.94
5.30
5.64
53.00
0.90
WA73-1-D-2-W4-5
16
10.78
5.29
5.49
51.00
0.88
3x3
WA73-1-D-2-W5-1
8
11.10
4.66
6.44
71.00
0.72
0.68
0.06
WA73-1-D-2-W5-2
10
11.21
4.94
6.27
78.00
0.70
WA73-1-D-2-W5-3
15
11.22
4.87
6.35
73.00
0.71
WA73-1-D-2-W5-4
12
11.29
4.74
6.56
75.00
0.73
WA73-1-D-2-W5-5
17
12.27
7.09
5.18
73.00
0.58
NA: Not Available
65
-------
Table A-1-4-1: Task 1 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Vinyl Flooring
Wipe
Type
Size
ID
Random
Number
Assignment
Weight
Wipe
Initial
of Wet
s(g)
Final
Net
Weight
Loss (g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss (g/ft2)
Dispatch8 Hospital Cleaner Disinfectant Towels with Bleach/Vinyl Flooring
1x1
WA73-1-V-3-W1 -1
20
13.35
11.39
1.96
24.00
1.96
1.96
0.10
WA73-1-V-3-W1 -2
16
13.10
11.27
1.83
21.00
1.83
WA73-1-V-3-W1 -3
25
12.93
10.86
2.08
18.00
2.08
WA73-1-V-3-W1 -4
1
12.47
10.42
2.04
18.00
2.04
WA73-1-V-3-W1 -5
10
12.98
11.11
1.88
19.00
1.88
1.5x1.5
WA73-1-V-3-W2-1
13
12.82
9.91
2.91
33.00
1.29
1.28
0.05
WA73-1-V-3-W2-2
5
12.67
9.63
3.04
29.00
1.35
WA73-1-V-3-W2-3
19
12.87
10.06
2.81
28.00
1.25
WA73-1-V-3-W2-4
22
12.99
10.26
2.73
22.00
1.21
WA73-1-V-3-W2-5
6
12.99
10.04
2.95
25.00
1.31
2x2
WA73-1-V-3-W3-1
4
12.64
8.73
3.92
34.00
0.98
0.97
0.01
WA73-1-V-3-W3-2
11
12.98
9.09
3.89
31.00
0.97
WA73-1-V-3-W3-3
7
12.84
8.95
3.89
35.00
0.97
WA73-1-V-3-W3-4
12
12.95
9.14
3.81
37.00
0.95
WA73-1-V-3-W3-5
23
13.19
9.36
3.84
30.00
0.96
2.5x2.5
WA73-1-V-3-W4-1
14
13.56
8.88
4.68
36.00
0.75
0.76
0.02
WA73-1-V-3-W4-2
24
13.00
8.32
4.68
36.00
0.75
WA73-1-V-3-W4-3
2
13.09
8.43
4.65
38.00
0.74
WA73-1-V-3-W4-4
21
13.18
8.32
4.87
54.00
0.78
WA73-1-V-3-W4-5
8
13.46
8.51
4.95
50.00
0.79
3x3
WA73-1-V-3-W5-1
17
13.50
8.56
4.94
56.00
0.55
0.56
0.01
WA73-1-V-3-W5-2
3
13.06
7.99
5.08
57.00
0.56
WA73-1-V-3-W5-3
18
12.93
8.04
4.89
54.00
0.54
WA73-1-V-3-W5-4
15
13.37
8.24
5.13
58.00
0.57
WA73-1-V-3-W5-5
17
12.27
7.09
5.18
57.00
0.58
66
-------
Table A-1-4-2: Task 1 Results Dispatch® Hospital Cleaner Disinfectants Towels with Bleach/Glass
Wipe
Type
Size
ID
Random
Number
Assignment
Weight of Wet
Wipes (g)
Net
Weight
Loss (g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss
(g/ft2)
Initial
Final
WA73-1-G-3-W1-1
10
10.55
9.69
0.86
21.00
0.86
WA73-1-G-3-W1-2
17
11.12
10.27
0.85
20.00
0.85
1x1
WA73-1-G-3-W1-3
7
10.09
9.15
0.94
21.00
0.94
0.92
0.07
WA73-1-G-3-W1-4
5
11.01
10.04
0.98
19.00
0.98
WA73-1-G-3-W1-5
19
11.64
10.66
0.99
18.00
0.99
>
>
CO
WA73-1-G-3-W2-1
15
10.44
8.43
2.01
24.00
0.89
O
S
o
CO
cu
WA73-1-G-3-W2-2
13
11.68
9.50
2.18
24.00
0.97
1.5x1.5
WA73-1-G-3-W2-3
3
10.53
8.62
1.92
26.00
0.85
0.88
0.06
'1
(/>
cu
WA73-1-G-3-W2-4
18
11.76
9.86
1.90
24.00
0.84
WA73-1-G-3-W2-5
12
10.93
9.06
1.87
27.00
0.83
s
o
i—
(/>
c
CO
WA73-1-G-3-W3-1
23
10.95
8.38
2.56
32.00
0.64
WA73-1-G-3-W3-2
9
10.59
8.11
2.48
33.00
0.62
o
<22
c
2x2
WA73-1-G-3-W3-3
16
10.86
8.45
2.41
40.00
0.60
0.64
0.03
(/)
Q
WA73-1-G-3-W3-4
4
12.11
9.34
2.77
31.00
0.69
cu
c
CO
cu
o
"co
WA73-1-G-3-W3-5
24
12.25
9.64
2.61
34.00
0.65
WA73-1-G-3-W4-1
1
11.06
7.12
3.94
43.00
0.63
Q.
>
WA73-1-G-3-W4-2
11
11.91
7.85
4.06
45.00
0.65
I
2.5x2.5
WA73-1-G-3-W4-3
22
12.77
8.76
4.00
40.00
0.64
0.64
0.01
o
CO
Q_
WA73-1-G-3-W4-4
21
12.04
8.02
4.02
41.00
0.64
(/)
Q
WA73-1-G-3-W4-5
25
13.00
8.91
4.10
42.00
0.66
WA73-1-G-3-W5-1
2
11.81
7.65
4.16
60.00
0.46
WA73-1-G-3-W5-2
20
11.89
7.67
4.22
58.00
0.47
3x3
WA73-1-G-3-W5-3
14
10.88
6.79
4.09
59.00
0.45
0.48
0.05
WA73-1-G-3-W5-4
8
10.44
6.36
4.09
59.00
0.45
WA73-1-G-3-W5-5
17
12.27
7.09
5.18
61.00
0.58
67
-------
Table A-1-4-3: Task 1 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Melamine
Wipe
Type
Size
ID
Random
Number
Assignment
Weight of Wet
Wipes (g)
Net
Weight
Loss (g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss (g/ft2)
Initial
Final
Dispatch8 Hospital Cleaner Disinfectant Towels with Bleach/Melamine
1x1
WA73-1-M-3-W1 -1
24
12.51
11.29
1.21
19.00
1.21
1.21
0.04
WA73-1-M-3-W1 -2
13
10.55
9.38
1.17
21.00
1.17
WA73-1-M-3-W1 -3
5
13.94
12.76
1.19
19.00
1.19
WA73-1-M-3-W1 -4
25*
13.58
12.39
1.20
20.00
1.20
WA73-1-M-3-W1 -5
9
14.10
12.82
1.27
23.00
1.27
1.5x1.5
WA73-1-M-3-W2-1
4
12.92
10.90
2.02
30.00
0.90
0.92
0.03
WA73-1-M-3-W2-2
10
14.59
12.61
1.98
28.00
0.88
WA73-1-M-3-W2-3
14
10.90
8.85
2.05
32.00
0.91
WA73-1-M-3-W2-4
8
14.80
12.71
2.09
29.00
0.93
WA73-1-M-3-W2-5
12
14.81
12.64
2.17
28.00
0.96
2x2
WA73-1-M-3-W3-1
3
13.90
10.38
3.52
32.00
0.88
0.93
0.03
WA73-1-M-3-W3-2
18
12.97
9.28
3.69
35.00
0.92
WA73-1-M-3-W3-3
22
13.09
9.20
3.88
59.00
0.97
WA73-1-M-3-W3-4
15
13.25
9.52
3.73
37.00
0.93
WA73-1-M-3-W3-5
16
13.67
9.99
3.68
34.00
0.92
2.5x2.5
WA73-1-M-3-W4-1
23
13.54
9.49
4.05
40.00
0.65
0.67
0.01
WA73-1-M-3-W4-2
17
12.46
8.33
4.13
39.00
0.66
WA73-1-M-3-W4-3
11
14.02
9.77
4.26
43.00
0.68
WA73-1-M-3-W4-4
1
12.80
8.56
4.24
42.00
0.68
WA73-1-M-3-W4-5
21*
12.69
8.46
4.23
44.00
0.68
3x3
WA73-1-M-3-W5-1
20
13.05
7.52
5.53
56.00
0.61
0.62
0.03
WA73-1-M-3-W5-2
19
13.90
8.21
5.69
54.00
0.63
WA73-1-M-3-W5-3
7
14.35
8.53
5.82
54.00
0.65
WA73-1-M-3-W5-4
2
13.29
7.57
5.72
61.00
0.64
WA73-1-M-3-W5-5
17
12.27
7.09
5.18
59.00
0.58
68
-------
Table A-1-4-4: Task 1 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Painted Drywall
Wipe
Type
Size
ID
Random
Number
Assignment
Weight of Wet
Wipes (g)
Net
Weight
Loss (g)
Net Wipe
Time
(seconds)
Net
Weight
Loss
(g/ft2)
Average
Weight
Loss
(g/ft2)
StDev
Weight
Loss (g/ft2)
Initial
Final
Dispatch8 Hospital Cleaner Disinfectant Towels with Bleach/Painted Drywall
1x1
WA73-1-D-3-W1-1
17
13.90
11.96
1.94
22.00
1.94
2.07
0.09
WA73-1-D-3-W1-2
11
13.90
11.86
2.04
25.00
2.04
WA73-1-D-3-W1-3
13
13.56
11.44
2.12
28.00
2.12
WA73-1-D-3-W1-4
2
14.01
11.88
2.13
26.00
2.13
WA73-1-D-3-W1-5
15
14.04
11.90
2.15
25.00
2.15
1.5x1.5
WA73-1-D-3-W2-1
22
14.14
10.63
3.51
30.00
1.56
1.62
0.05
WA73-1-D-3-W2-2
8
13.80
10.20
3.61
31.00
1.60
WA73-1-D-3-W2-3
3
13.92
10.12
3.80
29.00
1.69
WA73-1-D-3-W2-4
7
14.27
10.69
3.57
31.00
1.59
WA73-1-D-3-W2-5
12
14.09
10.39
3.70
31.00
1.64
2x2
WA73-1-D-3-W3-1
19
14.10
10.04
4.06
39.00
1.02
1.07
0.07
WA73-1-D-3-W3-2
25
13.96
9.93
4.03
44.00
1.01
WA73-1-D-3-W3-3
23
13.87
9.69
4.18
41.00
1.04
WA73-1-D-3-W3-4
16
14.07
9.48
4.59
46.00
1.15
WA73-1-D-3-W3-5
5
14.03
9.51
4.52
43.00
1.13
2.5x2.5
WA73-1-D-3-W4-1
6
14.11
8.54
5.57
62.00
0.89
0.89
0.02
WA73-1-D-3-W4-2
14
13.75
8.27
5.48
57.00
0.88
WA73-1-D-3-W4-3
9
13.83
8.10
5.73
55.00
0.92
WA73-1-D-3-W4-4
4
13.79
8.21
5.58
59.00
0.89
WA73-1-D-3-W4-5
24
14.10
8.71
5.39
59.00
0.86
3x3
WA73-1-D-3-W5-1
21
13.80
7.22
6.58
60.00
0.73
0.70
0.07
WA73-1-D-3-W5-2
18
14.21
7.63
6.57
61.00
0.73
WA73-1-D-3-W5-3
20
14.37
7.54
6.83
63.00
0.76
WA73-1-D-3-W5-4
10
13.64
7.21
6.43
74.00
0.71
WA73-1-D-3-W5-5
17
12.27
7.09
5.18
62.00
0.58
69
-------
Test Results for Task 2: Surface Retention Times of Wipe Decontaminant Liquid
Table A-2-1-1: Task 2 Results Clorox Healthcare® Bleach Germicidal Wipe/Vinyl Flooring
Wipe
Type
Wait
time
(min)
ID
Weight of Wet
Wipes (g)
Net
Weight
Loss (g)
Weight of Dry
Wipes (g)
Net
Weight
Gain
(g)
Average
Amount of
Liquid
Recovered
g/ft2
StDev of the
Average
Liquid
Recovered
g/ft2
Initial
Final
Initial
Final
Clorox Healthcare® Bleach Germicidal
Wipe/Vinyl Flooring
0
WA73-2-V-1-D0-1
12.241
9.194
3.047
4.581
6.65
2.069
1.88
0.15
WA73-2-V-1-D0-2
12.003
9.438
2.565
4.802
6.566
1.764
WA73-2-V-1-D0-3
12.304
9.499
2.805
4.762
6.498
1.736
WA73-2-V-1-D0-4
12.019
9.394
2.625
4.592
6.594
2.002
WA73-2-V-1-D0-5
12.184
9.123
3.061
4.617
6.455
1.838
5
WA73-2-V-1-D5-1
11.973
9.223
2.75
4.668
5.899
1.231
1.26
0.17
WA73-2-V-1-D5-2
12.37
8.968
3.402
4.563
6.018
1.455
WA73-2-V-1-D5-3
12.041
9.107
2.934
4.845
5.88
1.035
WA73-2-V-1-D5-4
11.949
8.846
3.103
4.574
5.749
1.175
WA73-2-V-1-D5-5
12.176
9.275
2.901
4.7
6.104
1.404
Table A-2-1-2: Task 2 Results Clorox Healthcare® Bleach Germicidal Wipe/Glass
Wipe
Type
Wait
time
(min)
ID
Weight of Wet
Wipes (g)
Net Weight
Loss (g)
Weight of Dry
Wipes (g)
Net Weight
Gain (g)
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
Initial
Final
Initial
Final
Clorox Healthcare® Bleach Germicidal
Wipe/Glass
0
WA73-2-G-1-D0-1
12.817
11.195
1.622
4.576
5.684
1.108
1.21
0.08
WA73-2-G-1-D0-2
11.382
9.398
1.984
4.806
6.123
1.317
WA73-2-G-1-D0-3
11.978
9.957
2.021
4.539
5.774
1.235
WA73-2-G-1-D0-4
12.169
10.378
1.791
4.672
5.813
1.141
WA73-2-G-1-D0-5
11.54
9.641
1.899
4.769
5.998
1.229
5
WA73-2-G-1-D5-1
11.75
10.23
1.52
4.555
4.93
0.375
0.37
0.09
WA73-2-G-1-D5-2
11.94
9.9
2.04
4.545
5.061
0.516
WA73-2-G-1-D5-3
12.144
10.626
1.518
4.602
4.979
0.377
WA73-2-G-1-D5-4
12.234
10.218
2.016
4.658
4.945
0.287
WA73-2-G-1-D5-5
11.657
9.602
2.055
4.717
5.009
0.292
70
-------
Table A-2-1-3: Task 2 Results Clorox Healthcare® Bleach Germicidal Wipe/Melamine
Wipe
Type
Wait
time
(min)
ID
Weight of Wet
Wipes (g)
Net
Weight
Loss (g)
Weight of Dry
Wipes (g)
Net Weight
Gain (g)
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
Initial
Final
Initial
Final
Clorox Healthcare® Bleach Germicidal
Wipe/Melamine
0
WA73-2-M-1-D0-1
11.446
9.553
1.893
4.594
5.811
1.217
1.29
0.12
WA73-2-M-1-D0-2
12.851
10.68
2.171
4.582
6.038
1.456
WA73-2-M-1-D0-3
12.227
10.465
1.762
4.609
5.942
1.333
WA73-2-M-1-D0-4
12.018
10.014
2.004
4.578
5.896
1.318
WA73-2-M-1-D0-5
11.764
9.753
2.011
4.599
5.747
1.148
5
WA73-2-M-1-D5-1
11.773
9.832
1.941
4.585
5.082
0.497
0.45
0.05
WA73-2-M-1-D5-2
12.259
10.383
1.876
4.574
5.03
0.456
WA73-2-M-1-D5-3
11.816
9.782
2.034
4.572
4.957
0.385
WA73-2-M-1-D5-4
12.422
10.633
1.789
4.601
5.115
0.514
WA73-2-M-1-D5-5
12.3
10.156
2.144
4.587
5.005
0.418
Table A-2-1-4: Task 2 Results Clorox Healthcare® Bleach Germicidal Wipe/Painted Drywall
Wipe
Type
Wait
time
(min)
Weight of Wet Wipes
(g)
Net
Weight of Dry
Wipes (g)
Net Weight
Gain (g)
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
ID
Initial
Final
Weight
Loss (g)
Initial
Final
Clorox Healthcare® Bleach Germicidal
WiDe/Painted Drvwall
0
WA73-2-D-1-D0-1
12.729
10.198
2.531
4.599
5.958
1.359
1.26
0.17
WA73-2-D-1-D0-2
12.278
10.152
2.126
4.592
5.668
1.076
WA73-2-D-1-D0-3
11.836
9.808
2.028
4.498
5.637
1.139
WA73-2-D-1-D0-4
12.076
9.686
2.39
4.514
6.013
1.499
WA73-2-D-1-D0-5
11.668
9.421
2.247
4.531
5.742
1.211
5
WA73-2-D-1-D5-1
12.24
10.249
1.991
4.624
4.971
0.347
0.33
0.03
WA73-2-D-1-D5-2
11.131
9.55
1.581
4.594
4.916
0.322
WA73-2-D-1-D5-3
12.212
10.137
2.075
4.612
4.909
0.297
WA73-2-D-1-D5-4
11.74
9.562
2.178
4.587
4.881
0.294
WA73-2-D-1-D5-5
11.964
9.81
2.154
4.579
4.944
0.365
71
-------
Table A-2-2-1: Task 2 Results Hype-Wipe® Bleach Towelette/Vinyl Flooring
Wipe
Type
Wait
time
(min)
ID
Weight of Wet Wipes
(g)
Net
Weight
Loss (g)
Weight of Dry
Wipes (g)
Net Weight
Gain (g)
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
Initial
Final
Initial
Final
Hype-Wipe® Bleach Towelette/Vinyl
Flooring
0
WA73-2-V4-D0-1
16.952
13.454
3.498
4.688
7.341
2.653
2.32
0.23
WA73-2-V4-D0-2
17.301
14.118
3.183
4.728
6.95
2.222
WA73-2-V4-D0-3
17.113
13.871
3.242
4.721
7.098
2.377
WA73-2-V4-D0-4
17.261
14.165
3.096
4.684
6.976
2.292
WA73-2-V4-D0-5
17.008
13.433
3.575
4.712
6.748
2.036
5
WA73-2-V4-D5-1
17.056
13.721
3.335
4.729
6.847
2.118
2.18
0.12
WA73-2-V4-D5-2
17.034
13.913
3.121
4.734
6.945
2.211
WA73-2-V4-D5-3
17.097
13.847
3.25
4.724
6.766
2.042
WA73-2-V4-D5-4
16.966
13.611
3.355
4.709
6.897
2.188
WA73-2-V4-D5-5
17.233
14.084
3.149
4.718
7.073
2.355
Table A-2-2-2: Task 2 Results Hype-Wipe® Bleach Towelette/Glass
Wipe
Type
Wait
time
(min)
ID
Weight of Wet Wipes
(g)
Net
Weight
Loss (g)
Weight of Dry
Wipes (g)
Net Weight
Gain (g)
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
Initial
Final
Initial
Final
V)
l/>
(0
X
5
WA73-2-G-4-D5-1
17.272
13.694
3.578
4.699
6.314
1.615
1.63
0.06
WA73-2-G-4-D5-2
17.297
13.753
3.544
4.739
6.295
1.556
WA73-2-G-4-D5-3
17.324
13.819
3.505
4.676
6.283
1.607
WA73-2-G-4-D5-4
17.347
13.685
3.662
4.717
6.359
1.642
WA73-2-G-4-D5-5
17.368
13.73
3.638
4.729
6.45
1.721
72
-------
Table A-2-2-3: Task 2 Results Hype-Wipe® Bleach Towelette/Melamine
Wipe
Wait
time
(min)
ID
Weight of Wet Wipes
(g)
Net
Weight
Loss (g)
Weight of Dry
Wipes (g)
Net Weight
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Type
Initial
Final
Initial
Final
Gain (g)
Liquid
Recovered
g/ft2
CD
WA73-2-M-4-D0-1
17.095
13.889
3.206
4.707
6.458
1.751
'£
03
WA73-2-M-4-D0-2
17.117
14.212
2.905
4.677
6.593
1.916
CD
n
"a5
"cd
CD
O
1—
0
WA73-2-M-4-D0-3
16.296
13.541
2.755
4.709
6.275
1.566
1.77
0.16
WA73-2-M-4-D0-4
17.084
13.967
3.117
4.731
6.68
1.949
WA73-2-M-4-D0-5
17.173
14.306
2.867
4.714
6.388
1.674
o
03
WA73-2-M-4-D5-1
17.313
14.235
3.078
4.67
5.838
1.168
CO
WA73-2-M-4-D5-2
17.449
14.165
3.284
4.708
5.93
1.222
CD
Q_
5
WA73-2-M-4-D5-3
17.244
14.44
2.804
4.719
5.785
1.066
1.16
0.06
CD
Q_
WA73-2-M-4-D5-4
17.266
14.372
2.894
4.723
5.881
1.158
WA73-2-M-4-D5-5
17.195
14.108
3.087
4.707
5.913
1.206
Table A-2-2-4: Task 2 Results Hype-Wipe® Bleach Towelette/Painted Drywall
Wipe
Wait
time
(min)
ID
Weight of Wet Wipes
(g)
Net
Weight
Loss (g)
Weight of Dry
Wipes (g)
Net Weight
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Type
Initial
Final
Initial
Final
Gain (g)
Liquid
Recovered
g/ft2
WA73-2-D-4-D0-1
17.091
13.441
3.65
4.734
6.933
2.199
CD
C
WA73-2-D-4-D0-2
17.186
13.379
3.807
4.736
6.982
2.246
03
Q_
"35
0
WA73-2-D-4-D0-3
17.142
13.518
3.624
4.737
7.117
2.38
2.48
0.32
cd
CD
WA73-2-D-4-D0-4
17.29
13.578
3.712
4.738
7.314
2.576
°!
WA73-2-D-4-D0-5
17.092
13.166
3.926
4.736
7.716
2.98
o C?
TO Q
CD
WA73-2-D-4-D5-1
17.15
13.866
3.284
4.738
5.256
0.518
CQ
®
WA73-2-D-4-D5-2
17.222
13.672
3.55
4.718
5.551
0.833
Q_
5
WA73-2-D-4-D5-3
17.167
13.41
3.757
4.729
5.242
0.513
0.62
0.18
I
WA73-2-D-4-D5-4
17.154
13.371
3.783
4.731
5.163
0.432
WA73-2-D-4-D5-5
17.209
13.573
3.636
4.716
5.514
0.798
73
-------
Table A-2-3-1: Task 2 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Vinyl Flooring
Wipe
Type
Wait
time
(min)
ID
Weight of Wet Wipes
(g)
Net
Weight
Loss (g)
Weight of Dry
Wipes (g)
Net Weight
Gain (g)
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
Initial
Final
Initial
Final
Sani-Cloth8 Bleach Germicidal Disposable
Wipe/Vinyl Flooring
0
WA73-2-V-2-D0-1
13.543
11.385
2.158
4.698
6.236
1.538
0.11
WA73-2-V-2-D0-2
13.339
11.694
1.645
4.727
5.991
1.264
WA73-2-V-2-D0-3
13.468
11.758
1.71
4.699
6.228
1.529
1.44
WA73-2-V-2-D0-4
14.214
12.336
1.878
4.634
6.074
1.44
WA73-2-V-2-D0-5
12.939
11.053
1.886
4.725
6.159
1.434
5
WA73-2-V-2-D5-1
13.625
11.89
1.735
4.755
5.221
0.466
0.08
WA73-2-V-2-D5-2
14.465
12.349
2.116
4.688
5.304
0.616
WA73-2-V-2-D5-3
12.967
11.264
1.703
4.629
5.217
0.588
0.52
WA73-2-V-2-D5-4
14.139
12.259
1.88
4.704
5.123
0.419
WA73-2-V-2-D5-5
13.623
11.722
1.901
4.741
5.235
0.494
Table A-2-3-2: Task 2 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Glass
Wipe
Type
Wait
time
(min)
ID
Weight of Wet Wipes
(g)
Net
Weight
Loss
Weight of Dry
Wipes
Net
Weight
Gain
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
Initial
Final
Initial
Final
Sani-Cloth8 Bleach Germicidal Disposable
Wipe/Glass
0
WA73-2-G-2-D0-1
13.267
12.065
1.202
4.63
4.981
0.351
0.39
0.06
WA73-2-G-2-D0-2
13.536
12.274
1.262
4.637
5.084
0.447
WA73-2-G-2-D0-3
13.262
11.952
1.31
4.702
5.045
0.343
WA73-2-G-2-D0-4
14.075
12.708
1.367
4.708
5.166
0.458
WA73-2-G-2-D0-5
13.434
12.226
1.208
4.691
5.046
0.355
5
WA73-2-G-2-D5-1
14.05
12.85
1.2
4.665
4.704
0.039
0.04
0.01
WA73-2-G-2-D5-2
13.66
12.49
1.17
4.635
4.68
0.045
WA73-2-G-2-D5-3
13.117
11.89
1.227
4.687
4.722
0.035
WA73-2-G-2-D5-4
13.621
12.437
1.184
4.805
4.839
0.034
WA73-2-G-2-D5-5
13.846
12.614
1.232
6.753
6.78
0.027
74
-------
Table A-2-3-3: Task 2 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Melamine
Wipe
Type
Wait
time
(min)
ID
Weight of Wet Wipes
(g)
Net
Weight of Dry
Wipes
Net
Weight
Gain
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
Initial
Final
Weight
Loss
Initial
Final
Sani-Cloth8 Bleach Germicidal Disposable
Wipe/Melamine
0
WA73-2-M-2-D0-1
13.076
11.806
1.27
4.605
5.204
0.599
0.62
0.10
WA73-2-M-2-D0-2
13.085
11.796
1.289
4.652
5.426
0.774
WA73-2-M-2-D0-3
13.124
11.873
1.251
4.692
5.217
0.525
WA73-2-M-2-D0-4
13.113
11.865
1.248
4.712
5.279
0.567
WA73-2-M-2-D0-5
13.261
11.745
1.516
4.581
5.234
0.653
5
WA73-2-M-2-D5-1
13.065
11.805
1.26
4.745
4.79
0.045
0.07
0.02
WA73-2-M-2-D5-2
13.367
12.156
1.211
4.551
4.637
0.086
WA73-2-M-2-D5-3
12.966
11.519
1.447
4.639
4.724
0.085
WA73-2-M-2-D5-4
13.079
11.664
1.415
4.65
4.728
0.078
WA73-2-M-2-D5-5
13.002
11.709
1.293
4.617
4.671
0.054
Table A-2-3-4: Task 2 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Painted Drywall
Wipe
Type
Wait
time
(min)
ID
Weight of Wet Wipes
(g)
Net
Weight
Loss
Weight of Dry
Wipes
Net
Weight
Gain
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the Average
Liquid
Recovered
g/ft2
Initial
Final
Initial
Final
Sani-Cloth8 Bleach Germicidal Disposable
Wipe/Painted Drywall
0
WA73-2-D-2-D0-1
13.517
11.965
1.552
4.61
5.523
0.913
0.86
0.09
WA73-2-D-2-D0-2
13.194
11.857
1.337
4.588
5.353
0.765
WA73-2-D-2-D0-3
12.892
11.591
1.301
4.616
5.607
0.991
WA73-2-D-2-D0-4
12.769
11.493
1.276
4.637
5.486
0.849
WA73-2-D-2-D0-5
13.427
11.836
1.591
4.663
5.469
0.806
5
WA73-2-D-2-D5-1
13.66
12.179
1.481
4.622
4.633
0.011
0.01
0.00
WA73-2-D-2-D5-2
13.139
11.81
1.329
4.598
4.605
0.007
WA73-2-D-2-D5-3
13.718
12.184
1.534
4.551
4.563
0.012
WA73-2-D-2-D5-4
13.32
11.819
1.501
4.614
4.622
0.008
WA73-2-D-2-D5-5
13.336
11.793
1.543
4.582
4.594
0.012
75
-------
Table A-2-4-1: Task 2 Results Dispatch® Hospital Cleaner Disinfectants Towels with Bleach/Vinyl Flooring
Wipe
Type
Wait
time
(min)
ID
Weight of Wet Wipes
(g)
Net
Weight
Weight of Dry
Wipes (g)
Net
Weight
Gain (g)
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
Initial
Final
Loss
(g)
Initial
Final
Dispatch8 Hospital Cleaner Disinfectants
Towels with Bleach/Vinyl Flooring
0
WA73-2-V-3-D0-1
13.909
11.911
1.998
4.567
6.084
1.517
1.48
0.12
WA73-2-V-3-D0-2
13.447
11.478
1.969
4.548
5.934
1.386
WA73-2-V-3-D0-3
12.172
10.542
1.63
4.57
5.928
1.358
WA73-2-V-3-D0-4
12.915
11.211
1.704
4.607
6.099
1.492
WA73-2-V-3-D0-5
13.244
11.408
1.836
4.523
6.188
1.665
5
WA73-2-V-3-D5-1
12.301
10.496
1.805
4.54
5.679
1.139
1.04
0.08
WA73-2-V-3-D5-2
12.713
11.063
1.65
4.591
5.682
1.091
WA73-2-V-3-D5-3
13.117
11.135
1.982
4.581
5.547
0.966
WA73-2-V-3-D5-4
13.093
11.102
1.991
4.568
5.513
0.945
WA73-2-V-3-D5-5
12.924
11.05
1.874
4.567
5.615
1.048
Table A-2-4-2: Task 2 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Glass
Wipe
Type
Wait
time
(min)
ID
Weight of Wet Wipes
(g)
Net
Weight
Loss
(g)
Weight of Dry
Wipes (g)
Net
Weight
Gain (g)
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
Initial
Final
Initial
Final
Dispatch8 Hospital Cleaner Disinfectant
Towels with Bleach/Glass
0
WA73-2-G-3-D0-1
12.791
11.935
0.856
4.566
4.903
0.337
0.35
0.02
WA73-2-G-3-D0-2
12.225
11.382
0.843
4.573
4.897
0.324
WA73-2-G-3-D0-3
12.481
11.553
0.928
4.587
4.935
0.348
WA73-2-G-3-D0-4
12.771
11.884
0.887
4.493
4.861
0.368
WA73-2-G-3-D0-5
13.046
12.082
0.964
4.601
4.982
0.381
5
WA73-2-G-3-D5-1
12.939
11.876
1.063
4.512
4.694
0.182
0.22
0.03
WA73-2-G-3-D5-2
13.008
12.086
0.922
4.497
4.759
0.262
WA73-2-G-3-D5-3
12.443
11.364
1.079
4.571
4.774
0.203
WA73-2-G-3-D5-4
12.3
11.491
0.809
4.562
4.803
0.241
WA73-2-G-3-D5-5
12.541
11.519
1.022
4.505
4.707
0.202
76
-------
Table A-2-4-3: Task 2 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Melamine
Wipe
Type
Wait
time
(min)
ID
Weight of Wet Wipes
(g)
Net
Weight
Loss
(g)
Weight of Dry
Wipes (g)
Net
Weight
Gain (g)
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
Initial
Final
Initial
Final
Dispatch8 Hospital Cleaner Disinfectant
Towels with Bleach/Melamine
0
WA73-2-M-3-D0-1
13.678
11.775
1.903
4.832
5.763
0.931
1.02
0.09
WA73-2-M-3-D0-2
13.012
10.936
2.076
4.764
5.686
0.922
WA73-2-M-3-D0-3
12.765
10.866
1.899
4.798
5.822
1.024
WA73-2-M-3-D0-4
13.101
11.428
1.673
4.589
5.711
1.122
WA73-2-M-3-D0-5
12.627
11.099
1.528
4.825
5.934
1.109
5
WA73-2-M-3-D5-1
12.953
11.143
1.81
4.803
5.314
0.511
0.61
0.07
WA73-2-M-3-D5-2
13.041
11.456
1.585
4.73
5.416
0.686
WA73-2-M-3-D5-3
13.42
11.367
2.053
4.686
5.365
0.679
WA73-2-M-3-D5-4
13.228
11.551
1.677
4.712
5.31
0.598
WA73-2-M-3-D5-5
13.416
11.892
1.524
4.809
5.401
0.592
Table A-2-4-4: Task 2 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Painted Drywall
Wipe
Type
Wait
time
(min)
ID
Weight of Wet Wipes (g)
Net
Weight of Wet
Wipes (g)
Net
Weight
Gain (g)
Average
Amount of
Liquid
Recovered
g/ft2
StDev of
the
Average
Liquid
Recovered
g/ft2
Initial
Final
Loss (g)
Initial
Final
Dispatch8 Hospital Cleaner Disinfectant
Towels with Bleach/Painted Drywall
0
WA73-2-D-3-D0-1
12.719
10.93
1.789
4.581
5.427
0.846
0.86
0.14
WA73-2-D-3-D0-2
13.156
11.473
1.683
4.535
5.313
0.778
WA73-2-D-3-D0-3
12.942
11.394
1.548
4.618
5.647
1.029
WA73-2-D-3-D0-4
12.222
10.81
1.412
4.573
5.534
0.961
WA73-2-D-3-D0-5
13.721
12.132
1.589
4.564
5.258
0.694
5
WA73-2-D-3-D5-1
12.554
10.856
1.698
4.58
4.869
0.289
0.30
0.05
WA73-2-D-3-D5-2
13.209
11.692
1.517
4.594
4.937
0.343
WA73-2-D-3-D5-3
12.678
11.127
1.551
4.631
4.852
0.221
WA73-2-D-3-D5-4
12.511
11.025
1.486
4.609
4.928
0.319
WA73-2-D-3-D5-5
12.398
10.599
1.799
4.57
4.902
0.332
77
-------
Test Results for Task 3: Impact of Wiping Pressure
Table A-3-1-1: Task 3 Results Clorox Healthcare® Bleach Germicidal Wipe/Vinyl Flooring
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes (g)
Net Weight
Loss (g/ft2)
Average
Weight Loss
(g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Clorox Healthcare® Bleach Germicidal
Wipe/Vinyl Flooring
Slight
Pressure
WA73-3-V-1-P1-1
13.637
12.052
1.585
1.69
0.07
WA73-3-V-1-P1-2
12.481
10.815
1.666
WA73-3-V-1-P1-3
13.132
11.447
1.685
WA73-3-V-1-P1-4
12.598
10.835
1.763
WA73-3-V-1-P1-5
13.076
11.342
1.734
Higher
Pressure
WA73-3-V-1-P3-1
12.142
8.453
3.689
3.90
0.31
WA73-3-V-1-P3-2
12.524
8.396
4.128
WA73-3-V-1-P3-3
12.357
8.418
3.939
WA73-3-V-1-P3-4
12.817
8.839
3.978
WA73-3-V-1-P3-5
12.886
9.134
3.752
Table A-3-1-2: Task 3 Results Clorox Healthcare® Bleach Germicidal Wipe/Glass
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes
(g)
Net Weight
Loss (g/ft2)
Average
Weight Loss
(g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Clorox Healthcare® Bleach Germicidal
Wipe/Glass
Slight
Pressure
WA73-3-G-1-P1-1
13.253
12.168
1.085
1.20
0.08
WA73-3-G-1-P1-2
11.702
10.404
1.298
WA73-3-G-1-P1-3
12.806
11.655
1.151
WA73-3-G-1-P1-4
11.519
10.312
1.207
WA73-3-G-1-P1-5
13.087
11.849
1.238
Higher
Pressure
WA73-3-G-1-P3-1
12.355
8.421
3.934
4.01
0.32
WA73-3-G-1-P3-2
12.726
8.634
4.092
WA73-3-G-1-P3-3
12.821
8.835
3.986
WA73-3-G-1-P3-4
13.015
9.156
3.859
WA73-3-G-1-P3-5
12.624
8.439
4.185
78
-------
Table A-3-1-3: Task 3 Results Clorox Healthcare® Bleach Germicidal Wipe/Melamine
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes
(g)
Net Weight
Loss (g/ft2)
Average
Weight Loss
(g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Clorox Healthcare® Bleach Germicidal
Wipe/Melamine
Slight
Pressure
WA73-3-M-1-P1-1
11.91
10.063
1.847
1.95
0.11
WA73-3-M-1-P1-2
12.8
10.892
1.908
WA73-3-M-1-P1-3
12.939
10.857
2.082
WA73-3-M-1-P1-4
12.145
10.096
2.049
WA73-3-M-1-P1-5
12.873
10.991
1.882
Higher
Pressure
WA73-3-M-1-P3-1
12.927
8.395
4.532
4.43
0.35
WA73-3-M-1-P3-2
13.166
8.865
4.301
WA73-3-M-1-P3-3
13.197
8.784
4.413
WA73-3-M-1-P3-4
12.628
8.298
4.33
WA73-3-M-1-P3-5
12.835
8.265
4.57
Table A-3-1-4: Task 3 Results Clorox Healthcare® Bleach Germicidal Wipe/Painted Drywall
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes
(g)
Net Weight
Loss (g/ft2)
Average
Weight Loss
(g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Clorox Healthcare® Bleach Germicidal
Wipe/Painted Drywall
Slight
Pressure
WA73-3-D-1-P1-1
12.442
10.826
1.616
1.63
0.07
WA73-3-D-1-P1-2
12.265
10.740
1.525
WA73-3-D-1-P1-3
11.874
10.214
1.66
WA73-3-D-1-P1-4
13.246
11.519
1.727
WA73-3-D-1-P1-5
12.718
11.085
1.633
Higher
Pressure
WA73-3-D-1-P3-1
13.022
9.396
3.626
3.87
0.31
WA73-3-D-1-P3-2
13.103
9.163
3.940
WA73-3-D-1-P3-3
11.824
8.078
3.746
WA73-3-D-1-P3-4
13.109
9.113
3.996
WA73-3-D-1-P3-5
13.005
8.951
4.054
79
-------
Table A-3-2-1: Task 3 Results Hype-Wipe® Bleach Towelette/Vinyl Flooring
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes
(g)
Net Weight
Loss (g/ft2)
Average
Weight Loss
(g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Hype-Wipe8 Bleach Towelette/Vinyl Flooring
Slight
Pressure
WA73-3-V-4-P1 -1
17.302
13.902
3.4
3.49
0.10
WA73-3-V-4-P1 -2
17.215
13.722
3.493
WA73-3-V-4-P1 -3
17.327
13.923
3.404
WA73-3-V-4-P1 -4
17.376
13.735
3.641
WA73-3-V-4-P1 -5
17.297
13.807
3.49
Higher
Pressure
WA73-3-V-4-P3-1
17.451
11.297
6.154
6.03
0.35
WA73-3-V-4-P3-2
17.281
11.458
5.823
WA73-3-V-4-P3-3
17.466
11.324
6.142
WA73-3-V-4-P3-4
17.364
11.369
5.995
WA73-3-V-4-P3-5
17.292
11.277
6.015
Table A-3-2-2: Task 3 Results Hype-Wipe® Bleach Towelette/Glass
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes
(g)
Net Weight
Loss (g/ft2)
Average
Weight Loss
(g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Hype-Wipe8 Bleach Towelette/Glass
Slight
Pressure
WA73-3-G-4-P1-1
17.388
13.834
3.554
3.52
0.02
WA73-3-G-4-P1-2
17.297
13.805
3.492
WA73-3-G-4-P1-3
17.402
13.896
3.506
WA73-3-G-4-P1-4
17.258
13.743
3.515
WA73-3-G-4-P1-5
17.373
13.838
3.535
Higher
Pressure
WA73-3-G-4-P3-1
17.275
11.987
5.288
5.39
0.31
WA73-3-G-4-P3-2
17.354
11.862
5.492
WA73-3-G-4-P3-3
17.369
11.917
5.452
WA73-3-G-4-P3-4
17.28
11.879
5.401
WA73-3-G-4-P3-5
17.304
12.012
5.292
80
-------
Table A-3-2-3: Task 3 Results Hype-Wipe® Bleach Towelette/Melamine
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes
(g)
Net Weight
Loss (g/ft2)
Average
Weight Loss
(g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Hype-Wipe8 Bleach Towelette/Melamine
Slight
Pressure
WA73-3-M-4-P1-1
17.468
13.898
3.57
3.53
0.10
WA73-3-M-4-P1-2
17.369
13.925
3.444
WA73-3-M-4-P1-3
17.242
13.674
3.568
WA73-3-M-4-P1-4
17.227
13.824
3.403
WA73-3-M-4-P1-5
17.368
13.712
3.656
Higher
Pressure
WA73-3-M-4-P3-1
17.312
11.36
5.952
5.79
0.33
WA73-3-M-4-P3-2
17.359
11.565
5.794
WA73-3-M-4-P3-3
17.111
11.485
5.626
WA73-3-M-4-P3-4
17.573
11.563
6.01
WA73-3-M-4-P3-5
17.286
11.714
5.572
Table A-3-2-4: Task 3 Results Hype-Wipe® Bleach Towelette/Painted Drywall
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes (g)
Net Weight
Loss (g/ft2)
Average
Weight Loss
(g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Hype-Wipe8 Bleach Towelette/Painted Drywall
Slight
Pressure
WA73-3-D-4-P1-1
17.295
13.928
3.367
3.48
0.11
WA73-3-D-4-P1-2
17.266
13.892
3.374
WA73-3-D-4-P1-3
17.487
13.98
3.507
WA73-3-D-4-P1-4
17.346
13.758
3.588
WA73-3-D-4-P1-5
17.41
13.833
3.577
Higher
Pressure
WA73-3-D-4-P3-1
17.381
11.603
5.778
5.69
0.33
WA73-3-D-4-P3-2
17.392
11.622
5.770
WA73-3-D-4-P3-3
17.524
12.05
5.474
WA73-3-D-4-P3-4
17.317
11.489
5.828
WA73-3-D-4-P3-5
17.561
11.984
5.577
81
-------
Table A-3-3-1: Task 3 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Vinyl Flooring
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes (g)
Net Weight
Loss (g)
Average
Weight Loss
(g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Sani-Cloth8 Bleach Germicidal Disposable
Wipe/Vinyl Flooring
Slight
Pressure
WA73-3-V-2-P1 -1
13.255
12.111
1.144
1.09
0.04
WA73-3-V-2-P1 -2
12.916
11.852
1.064
WA73-3-V-2-P1 -3
13.01
11.908
1.102
WA73-3-V-2-P1 -4
13.144
12.113
1.031
WA73-3-V-2-P1 -5
12.83
11.743
1.087
Higher
Pressure
WA73-3-V-2-P3-1
12.876
9.497
3.379
3.52
0.27
WA73-3-V-2-P3-2
12.943
9.332
3.611
WA73-3-V-2-P3-3
13.086
9.437
3.649
WA73-3-V-2-P3-4
12.715
9.27
3.445
WA73-3-V-2-P3-5
12.833
9.318
3.515
Table A-3-3-2: Task 3 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Glass
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes (g)
Net Weight
Loss (g)
Average
Weight Loss
(g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Sani-Cloth8 Bleach Germicidal Disposable
Wipe/Glass
Slight
Pressure
WA73-3-G-2-P1-1
13.941
12.789
1.152
1.19
0.08
WA73-3-G-2-P1-2
13.697
12.564
1.133
WA73-3-G-2-P1-3
13.258
11.951
1.307
WA73-3-G-2-P1-4
14.076
12.853
1.223
WA73-3-G-2-P1-5
13.359
12.245
1.114
Higher
Pressure
WA73-3-G-2-P3-1
14.047
10.287
3.76
3.65
0.26
WA73-3-G-2-P3-2
13.603
10.182
3.421
WA73-3-G-2-P3-3
14.401
10.799
3.602
WA73-3-G-2-P3-4
13.693
10.02
3.673
WA73-3-G-2-P3-5
13.931
10.149
3.782
82
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Table A-3-3-3: Task 3 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Melamine
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes (g)
Net Weight
Average
Weight Loss
(g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Loss (g)
Sani-Cloth8 Bleach Germicidal Disposable
Wipe/Melamine
Slight
Pressure
WA73-3-M-2-P1-1
13.671
12.622
1.049
0.98
0.08
WA73-3-M-2-P1-2
13.789
12.887
0.902
WA73-3-M-2-P1-3
13.562
12.59
0.972
WA73-3-M-2-P1-4
14.118
13.217
0.901
WA73-3-M-2-P1-5
14.085
13.011
1.074
Higher
Pressure
WA73-3-M-2-P3-1
14.338
10.911
3.427
3.57
0.25
WA73-3-M-2-P3-2
14.234
10.533
3.701
WA73-3-M-2-P3-3
14.067
10.297
3.77
WA73-3-M-2-P3-4
13.934
10.481
3.453
WA73-3-M-2-P3-5
14.129
10.608
3.521
Table A-3-3-4: Task 3 Results Sani-Cloth® Bleach Germicidal Disposable Wipe/Painted Drywall
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes (g)
Net Weight Loss
(g)
Average
Weight
Loss (g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Sani-Cloth® Bleach Germicidal Disposable
Wipe/Painted Drywall
Slight
Pressure
WA73-3-D-2-P1-1
13.215
11.886
1.329
0.09
WA73-3-D-2-P1 -2
13.806
12.359
1.447
WA73-3-D-2-P1 -3
14.107
12.544
1.563
1.46
WA73-3-D-2-P1 -4
13.582
12.067
1.515
WA73-3-D-2-P1 -5
13.327
11.869
1.458
Higher
Pressure
WA73-3-D-2-P3-1
14.05
10.764
3.286
0.25
WA73-3-D-2-P3-2
13.569
10.231
3.338
WA73-3-D-2-P3-3
13.181
9.62
3.561
3.38
WA73-3-D-2-P3-4
13.538
10.397
3.141
WA73-3-D-2-P3-5
13.296
9.713
3.583
83
-------
Table A-3-4-1: Task 3 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Vinyl Flooring
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes (g)
Net Weight Loss
(g)
Average
Weight
Loss (g/ft2)
StDev
Weight Loss
(g/ft2)
Initial
Final
Dispatch® Hospital Cleaner Disinfectant Towels
with Bleach/Vinyl Flooring
Slight
Pressure
WA73-3-V-3-P1 -1
12.119
10.083
2.036
2.03
0.07
WA73-3-V-3-P1 -2
12.307
10.194
2.113
WA73-3-V-3-P1 -3
12.185
10.243
1.942
WA73-3-V-3-P1 -4
12.466
10.388
2.078
WA73-3-V-3-P1 -5
12.353
10.395
1.958
Higher
Pressure
WA73-3-V-3-P3-1
12.107
9.24
2.867
2.80
0.23
WA73-3-V-3-P3-2
12.408
9.607
2.801
WA73-3-V-3-P3-3
12.432
9.638
2.794
WA73-3-V-3-P3-4
12.448
9.719
2.729
WA73-3-V-3-P3-5
12.124
9.302
2.822
Table A-3-4-2: Task 3 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Glass
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes (g)
Net Weight Loss
(g)
Average
Weight
Loss (g/ft2)
StDev
Weight
Loss (g/ft2)
Initial
Final
Dispatch8 Hospital Cleaner Disinfectant
Towels with Bleach/Glass
Slight
Pressure
WA73-3-G-3-P1-1
11.585
10.545
1.04
1.00
0.07
WA73-3-G-3-P1-2
11.566
10.644
0.922
WA73-3-G-3-P1-3
11.683
10.648
1.035
WA73-3-G-3-P1-4
11.952
11.031
0.921
WA73-3-G-3-P1-5
10.874
9.813
1.061
Higher
Pressure
WA73-3-G-3-P3-1
11.437
9.429
2.008
2.20
0.19
WA73-3-G-3-P3-2
11.761
9.443
2.318
WA73-3-G-3-P3-3
12.038
9.684
2.354
WA73-3-G-3-P3-4
11.639
9.506
2.133
WA73-3-G-3-P3-5
11.371
9.16
2.211
84
-------
Table A-3-4-3: Task 3 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Melamine
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes (g)
Net Weight
Loss (g)
Average
Weight Loss
(g/ft2)
StDev
Weight
Loss (g/ft2)
Initial
Final
Dispatch8 Hospital Cleaner Disinfectant
Towels with Bleach/Melamine
Slight
Pressure
WA73-3-M-3-P1 -1
11.548
10.334
1.214
1.16
0.06
WA73-3-M-3-P1-2
11.674
10.486
1.188
WA73-3-M-3-P1-3
12.415
11.281
1.134
WA73-3-M-3-P1-4
11.971
10.893
1.078
WA73-3-M-3-P1-5
12.104
10.897
1.207
Higher
Pressure
WA73-3-M-3-P3-1
11.332
8.589
2.743
2.75
0.23
WA73-3-M-3-P3-2
12.541
9.637
2.904
WA73-3-M-3-P3-3
12.31
9.784
2.526
WA73-3-M-3-P3-4
11.856
9.178
2.678
WA73-3-M-3-P3-5
12.493
9.606
2.887
Table A-3-4-4: Task 3 Results Dispatch® Hospital Cleaner Disinfectant Towels with Bleach/Painted Drywall
Wipe
Type
Pressure
Applied
ID
Weight of Wet Wipes (g)
Net Weight
Loss (g)
Average
Weight Loss
(g/ft2)
StDev
Weight
Loss (g/ft2)
Initial
Final
Dispatch8 Hospital Cleaner Disinfectant
Towels with Bleach/Painted Drywall
Slight
Pressure
WA73-3-D-3-P1-1
12.611
10.721
1.89
1.96
0.18
WA73-3-D-3-P1-2
12.337
10.208
2.129
WA73-3-D-3-P1-3
13.083
10.932
2.151
WA73-3-D-3-P1-4
12.479
10.593
1.886
WA73-3-D-3-P1-5
13.102
11.374
1.728
Higher
Pressure
WA73-3-D-3-P3-1
12.424
9.161
3.263
3.22
0.25
WA73-3-D-3-P3-2
12.612
9.502
3.110
WA73-3-D-3-P3-3
13.111
9.81
3.301
WA73-3-D-3-P3-4
12.957
9.769
3.188
WA73-3-D-3-P3-5
12.866
9.637
3.229
85
-------
APPENDIX B - PHASE 2: Evaluation of Efficacy of Hypochlorite Wipes
Table B-1: Decontamination Test Results
Clorox Healthcare® Wipe Results
Test ID
Positive Controls
Test Coupons
Drip Wipes
Decon Wipes*
1
2
Test 1
Test 2
Test 3
Test 1
Test 2
Test 3
Test Test Test
1 2 3
5-73 C0D1
9.47E+06
1.03E+07
1.58E+05
5.95E+04
2.59 E+04
3.50E+02
1.52E+02
1.19E+02
ND
5-73 C0D2
7.55E+06
1.12E+07
6.72E+04
1.04E+05
1.55E+05
3.38E+02
4.41 E+02
9.24E+02
5-73 C0G1
6.09E+06
8.13E+06
3.99E+03
1.04E+04
2.19E+04
5.93E+01
6.48E+02
1.45E+02
5-73 C0G2
7.19E+06
5.91 E+06
3.03E+03
4.56E+03
2.41 E+03
2.03E+02
1.44E+02
9.71 E+01
5-73 C1D1
4.93E+07
5.84E+07
2.40E+03
2.72E+04
4.66E+02
6.30E+03
2.65E+02
6.16E+03
5-73 C1G1
2.38E+07
3.75E+07
6.75E+02
8.78E+03
5.04E+02
5.52E+01
2.73E+02
4.48E+02
5-73 C2D1
6.13E+07
8.91 E+07
4.62E+04
2.83E+05
2.02E+05
5.09E+04
1.57E+04
3.01 E+03
5-73 C2G1
4.14E+07
5.93E+07
7.12E+06
2.34E+04
1.39E+06
1.53E+04
NA
NA
5-73 C3D1
5.10E+07
3.75E+07
2.00E+06
1.33E+06
3.66E+05
3.34E+04
3.36E+04
5.83E+04
5-73 C3G1
1.22E+07
3.07E+07
2.51 E+02
4.37E+03
2.13E+02
4.71 E+04
1.60E+04
2.66E+03
5-73 C3D2
4.19E+07
3.15E+07
5.59E+04
7.27E+05
5.06E+04
4.31 E+03
2.87E+02
5.88E+03
5-73 C3G2
4.21 E+07
5.44E+07
1.06E+04
1.12E+04
2.92E+04
1.05E+04
5.18E+03
3.62E+04
Hype-Wipe® Bleach Towelette results
5-73 H0D1
6.02E+06
8.43E+06
8.34E+03
1.18E+04
3.22E+03
4.25E+01
3.99E+01
1.53E+02
ND
5-73 H0D2
1.01 E+07
1.11 E+07
1.13E+04
1.19E+04
1.88E+03
1.64E+02
1.16E+02
2.15E+02
5-73 H0G1
6.96E+06
5.20E+06
1.12E+02
1.14E+03
4.86E+01
4.90E+02
2.40E+02
1.12E+02
5-73 H0G2
6.68E+06
9.14E+06
1.20E+03
9.17E+03
1.02E+02
5.60E+01
5.94E+01
2.60E+02
5-73 H1D1
5.42E+07
3.66E+07
8.10E+02
8.77E+03
1.64E+02
3.89E+02
2.40E+02
1.41 E+02
5-73 H1G1
4.72E+07
1.62E+07
4.30E+02
2.73E+02
7.70E+01
1.75E+01
6.30E+00
8.69E+00
5-73 H2D1
5.09E+07
7.14E+07
1.20E+05
1.60E+03
4.48E+04
1.54E+03
2.45E+02
1.26E+02
5-73 H2G1
5.04E+07
3.83E+07
6.42E+04
2.01 E+03
1.67E+03
2.04E+04
4.34E+02
1.25E+03
5-73 H3D1
4.81 E+07
5.25E+07
8.51 E+02
1.60E+04
4.44E+04
3.62E+02
6.77E+04
8.40E+03
5-73 H3G1
3.67E+07
5.03E+07
2.51 E+04
2.06E+03
2.44E+04
2.11 E+04
1.36E+04
4.11 E+04
5-73 H3D2
3.67E+07
3.85E+07
1.21 E+06
1.43E+05
7.53E+04
2.24E+03
4.62E+02
3.02E+02
5-73 H3G2
3.70E+07
4.10E+07
1.78E+03
2.17E+03
1.96E+02
3.50 E+04
2.43E+05
6.74E+04
*: After a minimum of 24 h at 4 °C
NA: Not Available
ND: No Viable Spores Detected
86
-------
&EPA
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
------- |