NRMRL-RTP-460
EPA Contract EP-C-05-060/T056
December 2010
Environmental and Sustainable Technology
Evaluation: Mold-Resistant Lonseal Flooring -
Lonseal, Inc., Lonwood Natural
Prepared by:
RTI International
Microbiology Department
3040 Cornwallis Rd
Research Triangle Park, NC 27709
Telephone: 919-541-8018
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TABLE OF CONTENTS
Page
Acronyms and Abbreviations iii
Acknowledgments iv
1.0 Introduction 1
2.0 Verification Approach 3
2.1 Test Material 3
2.2 Test Methods and Procedures 4
2.2.1 Test Organisms 4
2.2.2 Static Chambers 4
2.2.3 Test Design 5
2.2.4 Sample Preparation and Inoculation 5
2.2.5 Calculation of Mold Resistance 6
2.3 Sustainability Indicators and Issues 6
3.0 Results 8
3.1 Mold Resistance 8
3.2 Emissions of VOCs and Formaldehyde 10
3.3 Sustainability Issues 10
4.0 Data Quality Assessment 11
5.0 References 12
APPENDICES
Page
Appendix A
VOCs and Formaldehyde Emissions Testing A -1
LIST OF FIGURES
Figure 1-1. Diagram illustrating the conditions required for fungal growth on a material 2
Figure 2-1. Front surface of material 3
Figure 2-2. Back surface of material 3
Figure 3-1. Log change in Aspergillus versicolor inoculated on the test material over 12 weeks on
reference material and Lonseal 8
Figure 3-2. Log change in naturally occurring fungi over 12 weeks on reference material and Lonseal. 8
LIST OF TABLES
Table 3-1. Logio CFUs for test material (Lonseal) and reference material (wood) on each test date
(Mean ± SD) 7
Table 3-2. Test results for VOCs and formaldehyde emissions from Lonseal 9
Table 4-1. Data quality objectives 10
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Acronyms and Abbreviations
ACH
air changes per hour
ADQ
audit of data quality
ASTM
American Society for Testing and Materials
AATCC
American Association of Textile Chemists and Colorists
aw
water activity
CFU
colony forming unit
DNPH
2,4-dinitrophenylhydrazine
DQO
data quality objective
EPA
U.S. Environmental Protection Agency
ESTE
environmental and sustainable technology evaluations
ERH
equilibrium relative humidity
ETV
environmental technology verification
g
gram(s)
GC/MS
gas chromatography/mass spectrometry
ISO
International Organization for Standardization
MC
moisture content
ML
microbiology laboratories
ML SOP
microbiology laboratory standard operating procedure
QA
quality assurance
QAM
quality assurance manager
QAPP
quality assurance project plan
QC
quality control
QMP
quality management plan
RH
relative humidity
RTI
Research Triangle Institute (RTI International)
sec
second(s)
SOP
standard operating procedure
spp
species
t
temperature in degrees Celsius
TOP
technical operating procedure
T/QAP
test/quality assurance plan
TSA
technical system audit
TVOC
total volatile organic compounds
VOCs
volatile organic compounds
Og
microgram(s)
Om
micrometer(s)
UL
Underwriters Laboratories
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ACKNOWLEDGMENTS
The authors acknowledge the support of all of those who helped plan and conduct the verification
activities. In particular, we would like to thank Dr. Timothy Dean, EPA's Project Manager, and Robert
Wright, EPA's Quality Assurance Manager, both of EPA's National Risk Management Research
Laboratory in Research Triangle Park, NC. We would also like to acknowledge the assistance and
participation of our stakeholder group for their input.
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1.0 INTRODUCTION
The U.S. Environmental Protection Agency's Office of Research and Development (EPA-ORD)
operates the Environmental and Sustainable Technology Evaluation (ESTE) Program to facilitate the
deployment of innovative technologies through performance verification and information dissemination.
The ESTE program is intended to increase the relevance of Environmental Technology Verification
(ETV) Program projects by responding to near-term needs identified by the U.S. EPA program and
regional offices.
The ESTE program involves a three step process. The first step is a technology category selection
process conducted by ORD. The second step involves selection of the project team and gathering of
project collaborators and stakeholders. Collaborators can include technology developers, vendors,
owners, and users. They support the project through funding, cost sharing, and technical support.
Stakeholders can include representatives of regulatory agencies, trade organizations relevant to the
technology, and other associated technical experts. The project team relies on stakeholder input to
improve the relevance, defensibility, and usefulness of project outcomes. Both collaborators and
stakeholders are critical to development of the project test and quality assurance plan (TQAP), the end
result of step two. Step three includes the execution of the verification and quality assurance and review
process for the final reports.
This ESTE project evaluated microbial resistant building materials. EPA's National Risk Management
Research Laboratory contracted with the Research Triangle Institute (RTI) to establish an ETV/ESTE
Program for microbial-resistant building materials. RTI convened a group of stakeholders representing
government and industry with knowledge and interest in the areas of mold resistant building materials.
The group met in May and July 2006 and recommended technologies to be tested. RTI then developed
(and EPA approved) the "Test/Quality Assurance Plan for Mold-Resistant Building Material Testing
The tests described in this report were conducted following this plan.
Fungal growth and the resulting contamination of building materials is a well-documented problem,
especially after the reports from New Orleans and the U.S. Gulf Coast post Hurricane Katrina.
However, contaminated materials have been recognized as important indoor fungal reservoirs for years.
For example, contamination with fungi has been associated with a variety of materials including carpet,
ceiling tile, gypsum board, wallpaper, flooring, insulation, and heating, ventilation and air conditioning
components2"5.
Exposure to fungi may result in respiratory symptoms of both the upper and lower respiratory tract such
as allergy and asthma6. Everyone is potentially susceptible. However, of particular concern are children
with their immature immune systems and individuals of all ages that are immunocompromised7'8.
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One approach to limiting exposure is to reduce the levels of fungi in the indoor space. For some
sensitive individuals, limiting exposure through avoidance is an effective control method; however,
avoidance is not always possible or practical. The investigation, development, and application of
effective source controls and strategies are essential to prevent fungal growth in the indoor environment.
Mold resistant building material is a potentially effective method of source control.
Figure 1-1 illustrates the combination of moisture and nutrients required for microbial growth on a
material. Sufficient nutrients for growth may be provided by the material itself or through the
accumulation of dust on or in the material. When sufficient nutrients are available, the ultimate
determinant for microbial growth is availability of water. The more hygroscopic a material (e.g.
wallboard) is, the more impact on the overall hygroscopicity the surface treatments may have.
A building is not a sterile environment, nor
should it be. In fact, a building is frequently a
reservoir for microorganisms. While many
different types of microorganisms occupy indoor
spaces, it is well-recognized that fungi can
colonize and amplify on a variety of building
materials if sufficient nutrients and moisture are
present. These contaminated materials are known
to be important indoor reservoirs. Fungal growth
on natural and fabricated building materials can
be a major source of respiratory disease in
humans. Commonly, sufficient nutrients are
available and water is usually the growth factor
most limiting the establishment and growth of
microbial populations. Sufficient moisture for
growth may become available through water
incursion from leaks and spills, condensation on
cold surfaces, or absorption or adsorption
directly from the indoor air. The amount of
water required is not large, and materials that
appear dry to cursory inspection may be capable
of supporting microorganism growth.
Nutrient
Moisture
Growth
Material
Figure 1-1. Diagram illustrating the conditions
required for fungal growth on a material.
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2.0 VERIFICATION APPROACH
The ESTE test program measured the mold resistance of Lonseal Lonwood Natural flooring. Since the
EPA program office wanted testing performed on mold-resistant building materials, and Lonseal
markets this flooring material as such, it was a good candidate for testing. Tests for emissions of VOCs
and formaldehyde were also performed. An overview of the emissions procedures is found in the
Appendix. The detailed test methods can be found in RTI's test/QA project plan1.
2.1 TEST MATERIAL
The following description of the product was provided by the vendor and was not verified.
Lonwood Natural flooring is a sheet vinyl product with an embossed wood-grain texture. Constructed in
multiple layers and embossed with distinctive wood grains, it is composed of resin, plasticizers, fillers,
and pigments. The co-calendered wear layer is formulated to provide maximum resistance to foot traffic
in most commercial and healthcare applications. The middle layer provides dimensional stability, sound-
absorbing properties, and resiliency under foot. The backing layer provides strength and stability of the
flooring and enhances the bonding strength of the adhesive. Mold resistance is conveyed by the addition
of a proprietary chemical as a top layer formulation that is applied to the surface of the sheet vinyl
through a calendering process. Figures 2-1 and 2-2 show the front and back surfaces of the material.
Figure 2-1. Front surface of material
Figure 2-2. Back surface of material
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2.2 TEST METHODS AND PROCEDURES
Mold resistance testing was performed following the guidelines outlined in ASTM 63299. This method
was developed as part of a more comprehensive project to apply indoor air quality engineering to
biocontamination in buildings. One of the primary goals was to develop a scientific basis for studying
indoor air biocontaminants. Available methods, including those from ASTM, AATCC, and UL, for
evaluating the resistance of a variety of materials to fungal growth were surveyed. Although the basic
principles were similar, a major concern was the way growth on the different materials was evaluated.
Although quantitative methods for inoculation were employed by most of the methods, all assessed
growth qualitatively as the endpoint. ASTM 63299 evaluates growth quantitatively as the endpoint. The
method has been successfully used to evaluate fungal resistance on a variety of materials including
ceiling tiles and HVAC duct materials 10"13.
2.2.1 Test Organisms
Selecting the "correct" test organism is critical to any test, therefore selection criteria were developed.
The selection criteria used to choose the appropriate test organisms for this study were:
(1) the reasonableness or likelihood of the test material being challenged by that particular organism
when in actual use, and
(2) that they cover the range of ERHs (equilibrium relative humidities) needed and bracket the ERHs
where fungal growth can occur.
Two fungi were used as test organisms, Aspergillus versicolor and Stachybotrys chartarum. Each of
them met the criteria. S. chartarum requires high levels of available water to grow and has been
associated with a number of toxigenic symptoms. A. versicolor is a xerophilic fungus and capable of
growing at lower relative humidities. Both are from the RTI culture collection (CC). The CC number
for S. chartarum is 3075 and the organism was received from EPA NERL. A. versicolor is CC #3348,
and it is a field isolate. Prior to initiation of the testing, their identification was confirmed by standard
techniques.
2.2.2 Static Chambers
Clear plastic desiccators served as the static environmental chambers. The desiccators are sealed so there
is no air exchange and the desiccators serve as good static chambers. A saturated-salt solution of
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potassium chloride was used to maintain the humidity of the 85% ERH chamber. Sterile water was used
for the 100% ERH chamber. Temperature was externally controlled and maintained at room
temperature. Prior to use, the chambers were decontaminated and characterized. The ERH in each
chamber was monitored with a hygrometer (Taylor model number 5565) that was placed inside the
chamber.
2.2.3 Test Design
The Lonwood Natural flooring was cut aseptically with a razor blade into small pieces (at least 4 cm x 4
cm). The material was not autoclaved or sterilized in any way prior to inoculation. Therefore, in
addition to the test organism inocula, any organisms naturally on both the top and bottom surfaces of the
material had the opportunity to grow if conditions were favorable for growth. The test organisms are
inoculated by pipette directly onto the surface of each test piece in sufficiently high numbers to provide
an adequate challenge, but at a level that is realistic to quantify. The tests ran for 12 weeks. During the
12 week test period, data from four test dates, labeled Day 0, Week 1, Week 6, and Week 12 were
evaluated. Day 0 samples provided the baseline inoculum level. A sufficient number of test pieces were
inoculated simultaneously for all four test dates. All pieces for one material and one test organism were
put in the same static chamber. The chambers were set to 100% equilibrium relative humidity (ERH) for
the tests with S. chartarum and at 85% for A versicolor. On each test date (including Day 0), five
replicates of the test material pieces were removed from the chamber, each was placed separately in a
container with sterile buffer, and extracted by shaking. The resulting suspension of eluted organisms
was plated and microbial growth on materials was quantified by manually enumerating colony-forming
units (CFUs).
The numbers of CFUs eluted on week 1, 6, and 12 were compared to the baseline at Day 0. The
numbers of CFUs on each date are expressed as logio. The results are reported as the log change in
CFUs between Day 0 and Week 1, Day 0 and Week 6, and Day 0 and Week 12. An extra test date was
included to enable the QA review. The review had been scheduled for week 6, however scheduling
difficulties made the review impossible on week 6 so additional samples were processed on week 7 for
the audit.
2.2.4 Sample Preparation and Inoculation
Small (at least 4 cm x 4 cm) replicate pieces of test mold resistant flooring material and reference wood
material were prepared and inoculated. To minimize error and demonstrate reproducibility, five pieces
of each sample type were processed on each sampling date. Because there were four test dates, a
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minimum of 20 pieces were prepared simultaneously. Each piece was placed on a separate labeled
sterile Petri dish.
The fungi challenge suspensions were prepared by inoculating the test organism onto solid agar media,
incubating the culture at room temperature until mature, wiping organisms from the surface of the pure
culture, and suspending them in sterile 18-Mohm distilled water. The organism preparation was viewed
microscopically to verify purity of spores (absence of hyphae). The test pieces were inoculated (usually
with five 10 |iL spots in an X configuration) by pipet onto the surface of the test piece and allowed to
dry in the biosafety cabinet.
On each test date (including Day 0), the appropriate number of test pieces were removed from the static
chamber, each placed in approximately 30 mL sterile buffer, and extracted by shaking using a vortex or
wrist action shaker. The extract was diluted if needed and plated on agar media to determine the
numbers of CFU.
2.2.5 Calculation of Mold Resistance
Changes in the numbers of CFU over time were quantified. The logio number of CFUs from test date x
were compared to the logio number of CFU from Day 0 as follows:
A logio CFU = logw CFU^ate x - logw CFUDay 0
where:
A CFU = the change in log10 CFU between a test date (x) and Day 0
logw CFU(-[ate x = number of CFU logio on test date x
logw CFUDay 0 = number of CFU logio on Day 0
The standard error of the means between the start date and the test date gives the statistical significance
of the differences.
2.3 SU STAIN ABILITY INDICATORS AND ISSUES
The verification organization requested information from the vendor that would, along with the test
results for microbial resistance, assist in estimating impacts on solid waste disposal due to replacing
building materials less frequently. Information was also requested on chemical additives that are
claimed to confer microbial resistance. Also, the vendor was asked to provide any additional
information relative to the environmental sustainability of the product such as recyclability/reusability of
the product and disposability of the product and use of renewable resources or other criteria the vendor
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deemed relevant to the environmental sustainability of the product.
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3.0 RESULTS
3.1 MOLD RESISTANCE
The results for the mold resistance tests are shown in Table 3-1. Growth is measured by culture and is
defined as at least a 1 logio increase in culturable organism over the baseline which was determined on
Day 0.
Table 3-1. Log10 CFUs for test material (Lonseal) and reference material (wood) on each test date
(Mean ± SD)
Lonseal
Week
A. versicolor
85% ERH
S. chartarum
100% ERH
Growth of Naturally
Occurring Fungi
100% ERH
0
5.0 ±0.1
5.0 ±0.04
< 2.2 ± 0.0*
1
4.8 ±0.1
NA
4.8 ±0.6
6
4.4 ±0.1
NA
6.0 ±0.1
7
4.2 ±0.01
NA
6.2 ±0.2
12
4.1 ±0.1
NA
6.4 ±0.3
Reference Material
Week
A. versicolor
85% ERH
S. chartarum
100% ERH
Growth of Naturally
Occurring Fungi
100% ERH
0
4.9 ±0.1
4.8 ±0.1
< 2.2 ± 0.0*
1
4.7 ±0.1
3.9 ±0.2
2.6 ± 1.0
6
4.3 ±0.2
NA
6.3 ±0.0
7
4.1 ±0.1
NA
7.0 ±0.2
12
5.5 ±0.4
NA
6.9 ±0.3
NA = Not Available due to overgrowth by innate fungi
* = <2.2 indicates 0 CFU detected at the minimum detection limit
The numbers of CFUs on each test and reference piece were Logio transformed and the mean and
standard deviation calculated. The initial concentration is in the row labeled week 0 (day 0 inoculum).
The results for the test organisms, A. versicolor and S. chartarum, are in columns two and three. The
fourth column gives the CFUs for the fungi that were on the unsterilized surface of the test material at
the initiation of the test.
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At Day 0 the numbers of naturally
occurring fungi were below the
detection limit on both the test and
the reference materials. However, the
growth of a variety of fungal species
(naturally occurring on the sample)
was masking any S. chartarum
growth on Lonseal and on the
reference material (wood).
Figure 3-1 shows the log change in A
versicolor and Figure 3-2 shows the
log change in the naturally occurring
fungi that were on the surface of the
material.
Neither the test material nor the
reference material inoculated with A.
versicolor and incubated at 85%
ERH showed growth during the 12
weeks of the test. It was important to
check that none of the changes made
to the test material to make it mold
resistant actually enhanced the
ability of mold to grow over the
positive control material11
It was not possible to accurately
assess whether or not the test
material was resistant to the growth
of S. chartarum. The growth of a
variety of fungal species (naturally
occurring on the sample) masked
any S. chartarum growth on Lonseal
and on the reference material.
Z)
LL
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3.2 EMISSIONS OF VOCs AND FORMALDEHYDE
The emissions of VOCs and formaldehyde test results are presented in the Table 3-2. A summary of the
method is found in Appendix A14.
Table 3-2. Test results for VOCs and formaldehyde emissions from Lonseal
VOCs and Formaldehyde Emissions*
Emission Types
Minimum emission results
Total VOCs
< 0.5 mg/m3
Formaldehyde
<0.1 ppm
Individual VOCs
< 0.1 TLV
Individual pollutants must produce an air concentration level no greater than 1/10 the threshold limit
value (TLV) industrial workplace standard (Reference: American Conference of Government Industrial
Hygienists, 6500 Glenway, Building D-7, Cincinnati, OH 45211-4438.
3.3 SUSTAINABILITY ISSUES
Sustainability is an important consideration in use of microbial resistant building materials. Lonseal
supplied the following information about the sustainability of the Lonwood Natural flooring:
• Part of GreenMedic™ Microbial Resistant Collection
• The MSDS that was provided by the vendor was in Japanese. The percentage of the proprietary
compound is 0.01-0.02%. It is a top layer formulation that gets applied to the surface of the
sheet vinyl through a calendering process. The compound is added to the formula in a mixer, and
the product gets kneaded until ready for calendering. The MSDS cannot be distributed.
• Over 40% post industrial recycled content; contributes toward LEED MR 4.1 and 4.2 for
recycled content
• Part of GreenAir™ Collection: reformulated products that reduce VOC emissions by 80-90
percent
• Certified for low VOCs by GREENGUARD Environmental Institute
• Indoor Environmental Quality Credit 4.1 for low emitting adhesives
• Lonseal has a dedicated vinyl products recycling plant.
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4.0 DATA QUALITY ASSESSMENT
The quality assurance officer has reviewed the test results and the quality control data and has concluded
that the data quality objectives given in the approved Test/QA plan and shown in Table 4 have been
attained.
The DQO for the critical measurement, quantitation of fungal growth on an individual test date, is found
in Table 4-1.
Table 4-1. Data quality objectives
Test
Mold
Resistance
Parameter
Quantitation of
fungal growth on
an individual test
date
DQO
Precision
Accuracy
Completeness
± 5-fold
difference
10% of the plates will
be counted by a
second operator.
± 20% agreement
between the operators
100%
This verification statement discusses two aspects of Mold-Resistant Building Material Testing, mold
resistance and emissions of VOCs and formaldehyde. Users of this technology may wish to consider
other performance parameters such as fire resistance, service life and cost when selecting a building
material.
According to the test/QA plan1, this verification statement is valid for three years following the last
signature added on the verification statement.
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5.0 REFERENCES
1. RTI (Research Triangle Institute). 2008. Test/QA Plan for Mold-Resistant Building Material
Testing. Research Triangle Park, NC. http://www.epa.gov/etv/este.html
2. Morey, P.R., 1988, "Microorganisms in Buildings and HVAC Systems: A Summary of 21
Environmental Studies," Proceedings of the ASHRAE Conference on Indoor Air Quality, American
Society of Heating, Refrigeration, and Air-Conditioning Engineers, Atlanta, GA, pp 10-24.
3. Reynolds, S.J„ A.J. Steifel, and C.E. McJilton, 1990, Elevated Airborne Concentration of Fungi in
Residential and Office Environments, American Industrial Hygiene Association Journal, Vol. 51,
pp 601-604.
4. Leese, K.E., E.C. Cole, and J.D. Neefus, 1992, Biocide Mitigation of a Mold Contaminated
Building: An Initial Preventive Approach, Proceedings, American Industrial Hygiene Association
Annual Meeting, Washington, DC.
5. Kozak, P.P., et al, 1980, Currently Available Methods for Home Mold Surveys. II. Examples of
Problem Homes Surveyed, Annals of Allergy, Vol. 45, pp 167-176.
6. Garrett, M.H., Rayment, P.R., Hooper, M.A., Abramson, M.J., and Hooper, B.M. 1998, Indoor
airborne fungal spores, house dampness and associations with environmental factors and respiratory
health in children, Clinical and Experimental Allergy. 28: 459-467.
7. Rylander, R. and Etzel, R., 1999, Indoor mold and children's health. Environmental Health
Perspectives Supplements: 107: 465-517.
8. Gent, J.F., Ren, P., Belanger, K., Triche, E., Bracken, M.B., Holford, T.R., and Leaderer, B.P.,
2002, Levels of household mold associated with respiratory symptoms in the first year of life in a
cohort at risk for asthma. Environmental Health Perspectives: 110: A781-A786.
9. ASTM D6329-98(2003), Standard Guide for Developing Methodology for Evaluating the Ability of
Indoor Materials to Support Microbial Growth Using Static Environmental Chambers, American
Society for Testing and Materials, West Conshohocken, PA.
10. Foarde, K.K. and M.Y. Menetrez. 2002, Evaluating the Potential Efficacy of Three
Antifungal Sealants of Duct Liner and Galvanized Steel as Used in HVAC Systems. Journal
of Industrial Microbiology & Biotechnology. 29:3 8-43.
11. Foarde, K.K. and J.T. Hanley. 2001, Determine the Efficacy of Antimicrobial Treatments of
Fibrous Air Filters. ASHRAE Transactions. Volume 107, Part 1. 156-170.
12. Chang, J.C.S., K.K. Foarde, and D.W. VanOsdell. 1995, Growth Evaluation of Fungi
(Penicillium and Aspergillus spp.) On Ceiling Tile. Atmospheric Environment. 29:2331
2337.
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13. Foarde, K., E. Cole, D. VanOsdell, D. Bush, D. Franke and J. Chang. 1992, Characterization of
Environmental Chambers for Evaluating Microbial Growth on Building Materials. In: IAQ '92
Environments for People, proceedings; 185-190.
14. ASTM. 2006. D5116-06, Standard Guide for Small Scale Environmental Chamber Determinations
of Organic Emissions from Indoor Materials/Products, American Society for Testing and
Materials, West Conshohocken, PA.
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Appendix A
VOCs and Formaldehyde Emissions Testing
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EMISSIONS REPORT FOR LONSEAL FLOORING MATERIAL
Two pieces of Lonseal flooring material, contained in a 7"x7"x2" cradle of aluminum foil, were tested
in the small (52.7 L capacity) emissions chamber maintained at 25 °C and 50% relative humidity and
subjected to an air exchange rate of 1 hr"1. After equilibration of each sample for 6 hr1, sequential
samples for VOCs and carbonyls were collected from the chamber effluent for 20 and 120 minutes,
yielding collection volumes of approximately 1.5 and 10 L for VOCs and 10 and 60 L for carbonyls2. In
addition to the test flooring material, replicate chamber blanks and the emission profile of a positive
control material (vinyl show curtain liner) were collected. All sample collections and analyses were
conducted in accordance with RTFs AIHA quality manual guidelines.3
VOC samples were collected on Carbopack B cartridges. A total of 100 ng of the internal standard, d8-
toluene, was subsequently added to each cartridge by flash loading4 prior to analysis by thermal
desorption-GC/MS on a DB-5 column programmed from 40EC - 225EC at 5 EC/min5. Calibration
standards were prepared at two levels (3.5 |ig; 6.9 ng) by flash loading of a 26-component VOC mixture
(ethanol; isopropanol; acetone; dichloromethane; carbon disulfide; methyl -t -butyl ether; 2-butanone;
1,1,1-trichloroethane; 1-butanol; trichloroethene, 4-methyl-2-pentanone; toluene; hexanal;
tetrachloroethene; m-xylene; n-nonane; 2-butoxyethanol; phenol; 1,2,4-trimethylbenzene; n-decane; 2-
ethyl-l-hexanol; d-limonene; 1,2-dichlorobenzene; n-undecane; decamethylcyclosiloxane; n-dodecane)
plus d9-toluene internal standard in methylene chloride onto Carbopack B. In addition to quantitation of
the individual analytes, total VOCs (TVOC) were determined by summing the integrated peak areas in
the samples and blanks between the retention times of hexane and hexadecane. Two specific analytes, 4-
phenylcyclohexene and styrene, were sought in each sample. Neither compound was detected in the
samples or blanks. All detected analytes were quantitated against the toluene peak in the standards. No
mathematical correction for the blanks was performed.
Carbonyl samples were collected on DNPH cartridges.2'6 Each cartridge was extracted by solid phase
extraction (SPE) with 4 mL of acetonitrile and brought to a final volume of 5 mL with acetonitrile7.
Subsequently, each extract was analyzed by HPLC/UV (365 nm) on a Deltabond Res AK column (4.6
mm x 25 cm, Keystone). The mobile phase consisted of (A) 45:55 acetonitrile:water and (B) 75:25
acetonitrile:water, using a 30 minute gradient from A to B and held at B for 5 minutes at a flow rate of 1
mL/min. Each cartridge was extracted by solid phase extraction (SPE) with 4 mL of acetonitrile and
brought to a final volume of 5 mL with acetonitrile. Instrument calibration was accomplished using
solutions prepared from a purchased aldehyde/ketone DNPH mix solution (15 |ag/mL as formaldehyde,
Supelco 47285-U) in acetonitrile. A six-point calibration curve was prepared with analyte amounts
ranging from 0.0109 to 2.175 |ig/ml. Individual carbonyls (formaldehyde, acetaldehyde, acetone,
proprionaldehyde, crotonaldehyde, butyraldehyde, benzaldehyde, iso-valeraldehyde, valeraldehyde, o-
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tolualdehyde, m-tolualdehyde, p-tolualdehyde, hexanaldehyde, 2,5-dimethylbenzaldehyde ) were
quantitated against the curve and were corrected for amounts found in blank samples. Total carbonyls
were computed by summing the individual carbonyl species.
The results of the emission tests for VOCs and carbonyls are presented in Tables 1 and 2, respectively.
For all samples, excluding the positive control, levels of VOCs and carbonyls were extremely small,
near the detection limit for the method, and comparable to the levels found in the blanks.
Table 1. VOC emission results3 for Lonseal flooring material
Sample Id.
Toluene
Chamber
Cone, (mg/m3)
TVOC
Chamber
Cone, (mg/m3)
Toluene
Emission
Factor
(mg/m2-hr)
TVOC
Emission
Factor
(mg/m2-hr)
Chamber Blankb
0.009 (0.005)
0.25 (0.116)
0.015 (0.008)
0.43 (0.20)
Positive Control0
0.017 (0.007)
14.2 (1.1)
0.029 (0.012)
23.6 (1.8)
Lonseal flooringd
0.003 (0.003)
0.27 (0.13)
0.006 (0.005)
0.46 (0.43)
a Mean (Standard deviation)
b Mean of 3 determinations
c Mean of 2 determinations
d Mean of 6 determinations
Table 2. Carbonyl emission results9 for Lonseal flooring material.
Sample Id.
Formaldehyde
Chamber
Cone, (mg/m3)
Total
Carbonyls
Chamber
Cone, (mg/m3)
Formaldehyde
Emission
Factor
(mg/m2-hr)
Total
Carbonyls
Emission
Factor
(mg/m2-hr)
Chamber Blankb
<0.001
0.017 (0.013)
<0.001
0.028 (0.023)
Positive Control15
<0.001
0.012 (0.013)
<0.001
0.021 (0.022)
Lonseal flooring0
0.001 (0.002)
0.015 (0.012)
0.003 (0.004)
0.026 (0.021)
a Mean (Standard deviation)
b Mean of 2 determinations
c Mean of 6 determinations
A - 3
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RTI International/EPA
December 2010
1 Standard Guide for Small-Scale Environmental Chamber Determinations of Organic Emissions from
Indoor Materials/Products. American Society for Testing and Materials (ASTM) document D5116-97,
2008.
2 Standard Operating Procedure for the Determination of Carbonyl and VOC Emissions from Building
Materials Using a Small Environmental Chamber. RTI International document: EAR-LAB-001, 2010.
3 Quality Manual for the AIHA Accredited Laboratory No. 100600. RTI International document:
RTI/0290365/08-01, January 2010.
4 Adsorbent Tube Injector System Operation Manual, Sigma-Aldrich/Supelco, Available at:
http://www.voungwha.com/tech/upload/ATIS system T702019.pdf 2010.
5 Standard Operating Procedure for the Analysis of Volatile Organic Chemicals By Thermal
Desorption/GC/MS, RTI International document: EAR-GLC-004, 2010.
6 Standard Test Method for Determination of Formaldehyde and Other Carbonyl Compounds in Air
(Active Sampler Methodology). American Society for Testing and Materials (ASTM) document D5197-
09, 2009.
7 Standard Operating Procedure for the Extraction and Analysis of Formaldehyde-DNPH from Active
and Passive Media by HPLC, RTI International document: EAR-GLC-003, 2010.
A - 4
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RTI International/EPA
September 2010
EPA Contract EP-C-05-060/T056
NRMRL-RTP-460
Environmental and Sustainable Technology
Evaluation: Mold-Resistant Lonseal Flooring -
Lonseal, Inc., Lonwood Natural
BRTI
INTERNATIONAL
Prepared by
Research Triangle Institute
jSsEPA
For
U.S. Environmental Protection Agency
Office of Research and Development- Environmental Technology Verification Program
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RTI International/EPA
September 2010
Page intentionally left for EPA Review Notice
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RTI International/EPA
September 2010
l lll ENVIRONMENTAL TECHNOLOGY VERIFICATION PROGRAM
Environmental and Sustainable Technology Evaluation (ESTE)
ElV
as EPA HRTI
' ~" INTERNATIONAL
U.S. Environmental Protection Agency
Research Triangle Institute
ESTE Joint Verification Statement
TECHNOLOGY TYPE:
Mold-Resistant Flooring Product
APPLICATION:
Flooring
TECHNOLOGY NAME:
Lonseal Lonwood Natural
COMPANY:
Lonseal, Inc.
ADDRESS:
Carson, California
The U.S. Environmental Protection Agency (EPA) has created the Environmental Technology
Verification (ETV) Program to facilitate the deployment of innovative or improved
environmental technologies through performance verification and dissemination of information.
The goal of the ETV Program is to further environmental protection and sustainability by
accelerating the acceptance and use of improved and cost-effective technologies. ETV seeks to
achieve this goal by providing high quality, peer-reviewed data on technology performance to
those involved in the purchase, design, distribution, financing, permitting, and use of
environmental technologies. This verification was conducted under the Environmental and
Sustainable Technology Evaluation (ESTE) element of the ETV Program that was designed to
address agency priorities for technology verification.
This ESTE project involved evaluation of the mold resistance of Lonseal Lonwood Natural
flooring. Tests for emissions of VOCs and formaldehyde were also performed. For this project
Research Triangle Institute (RTI) was the responsible contractor for EPA Office of Research and
Development, National Risk Management Research Laboratory (NRMRL).
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RT! International/EPA
September 2010
This verification statement provides a summary of the test results for Lonseal Lonwood Natural
flooring.
TECHNOLOGY DESCRIPTION
The following description of the product was provided by the vendor and was not verified.
Lonwood Natural flooring is a sheet vinyl product with an embossed wood-grain texture.
Constaicted in multiple layers and embossed with distinctive wood grains, it is composed of
resin, plasticizers, fillers, and pigments. The co-calendered wear layer is formulated to provide
maximum resistance to foot traffic in most commercial and health care applications. The middle
layer provides dimensional stability, sound-absorbing properties, and resiliency under foot. The
backing layer provides strength and stability of the flooring and enhances the bonding strength of
the adhesive. Mold resistance is conveyed by the addition of a proprietary chemical as a top layer
formulation that is applied to the surface of the sheet vinyl through a calendering process.
Figures S-l and S-2 show the front and back surfaces of the material.
Figure S-1. Front surface of material Figure S-2, Back surface of material
VERIFICATION TEST DESCRIPTION
Verification testing of the Lonseal Lonwood Natural flooring began on December 9, 2008 at the
microbiology laboratories of RTI International and was completed on March 3, 2009. All tests
were performed according to the ETV Program's "Test/QA Plan for Mold-Resistant Building
Material Testing." Mold resistance testing was performed following the guidelines outlined in
ASTM 6329. ASTM 6329 provides a quantitative endpoint for growth in a well-controlled, static
chamber environment. The method has been successfully used to evaluate fungal resistance on a
variety of materials including ceiling tiles and HVAC duct materials.
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RTI International/EPA
September 2010
In overview, the Lonwood Natural flooring sheet was cut aseptically with a razor blade into a
number of small test pieces (at least 4 cm x 4 cm). The material was not autoclaved or sterilized
prior to inoculation. Therefore, in addition to the test organism inocula, any organisms naturally
on both the top and bottom surfaces of the material had the opportunity to grow if conditions
were favorable for growth. The test organisms were inoculated by pipette directly onto the
surface of each test material piece in sufficiently high numbers to provide an adequate challenge,
but at a level that is realistic to quantify. The tests ran for 12 weeks. During the 12 week test
period, data from four test dates, labeled Day 0, Week 1, Week 6, and Week 12, were evaluated.
Day 0 samples provided the baseline inoculum level. A sufficient number of test pieces were
inoculated simultaneously for all four test dates. All pieces for one material and one test
organism were put in the same static chamber. Because Lonseal is a flooring material, the
reference material chosen for comparison was wood.
Two test organisms, Stachybotrys chartarum and Aspergillus versicolor were used. The static
chambers were set to 100% equilibrium relative humidity (ERH) for the tests with S. chartarum
and to 85% ERH for A. versicolor. On each test date (including Day 0), five replicates of the test
material pieces were removed from the chamber, each was placed separately in a container with
sterile buffer, and extracted by shaking. The resulting suspension of eluted organisms was plated
and microbial growth on materials was quantified by manually enumerating colony-forming
units (CFU).
The numbers of CFU eluted on test dates Weeks 1, 6, and 12 were compared to the baseline at
Day 0. The numbers of CFU were expressed as logi0. The results are reported as the logio
change in CFUs between Day 0 and Week 1, Day 0 and Week 6, and Day 0 and Week 12. An
extra test date was included to enable the QA review. The review had been scheduled for week 6,
however scheduling difficulties made the review impossible on week 6 so additional samples
were processed on week 7 for the audit.
Additional measurements included VOC and aldehyde emissions; these were performed by RTI
following ASTM D5116-06.
VERIFICATION OF PERFORMANCE
The results for the Mold Resistance tests are presented in the Figures S-3 and S-4. Growth is
measured by sporulation and is defined as at least a 1 logio increase in culturable organism over
the baseline which was determined on Day 0.
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RTI International/EPA
September 2010
Figure S-3 shows the log
change from the inocula on
Day 0 from A. versicolor and
Figure S-4 shows the log
change in the naturally
occurring fungi that were on
the surface of the material.
Neither the test material nor
the reference material
inoculated with A. versicolor
and incubated at 85% ERH
showed growth during the 12
weeks of the test. It was
important to check that none
of the changes made to the
test material to make it mold
resistant actually enhanced the ability of mold to grow over the reference material.
6.00 -|
5.00 -
4.00 -
Z)
3.00 -
o
2.00 -
d)
O)
c
1.00 -
03
o
0.00 -
O)
n
-1.00 -
_i
-2.00 J
|-h
'
6
Week
12
~ Reference Material ¦ Lonsea
Figure S-3. Log change in Aspergillus versicolor
inoculated on the test material over 12 weeks on
reference material and Lonseal.
It was not possible to accurately assess whether or not the test material was resistant to the
growth of S. char tar um. The
growth of a variety of fungal
species (naturally occurring on
the sample) masked any S.
chartarum growth on Lonseal
and on the reference material.
The quality assurance officer
reviewed the test results and
the quality control data and
concluded that the data quality
objectives given in the
approved test/QA plan were
attained.
-1.00
-2.00
Week
~ Reference Material ¦ Lonseal
Figure S-4. Log change in naturally occurring fungi
over 12 weeks on reference material and Lonseal.
The emissions of VOCs and
formaldehyde test results are
presented in Table S-l.
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RTI International/EPA
September 2010
Table S-l. Test results for VOCs and formaldehyde emissions from Lonseal
VOCs and Formaldehyde Emissions*
Emission Types
Minimum emission results
Total VOCs
< 0.5 mg/m3
Formaldehyde
<0.1 ppm
Individual VOCs
<0.1 TLV
individual pollutants must produce an air concentration level no greater than 1/10 the threshold limit
value (TLV) industrial workplace standard (Reference: American Conference of Government Industrial
Hygienists, 6500 Glenway, Building D-7, Cincinnati, OH 45211-4438.
This verification statement discusses two aspects of Mold-Resistant Building Material Testing,
mold resistance and emissions of VOCs and formaldehyde. Users of this technology may wish to
consider other performance parameters such as fire resistance, service life, and cost when
selecting a building material. According to the test/QA plan, this verification statement is valid
for 3 years following the last signature added on the verification statement.
Details of the verification test design, measurement test procedures, and Quality
Assurance/Quality Control Procedures can be found in the Test Plan titled Test OA Plan for
Mold-Resistant Building Material Testing (RTI 2008). Detailed results of the verification are
presented in the Final Report titled Environmental and Sustainable Technology Evaluation:
Mold-Resistant Lonseal Flooring - Lonseal, Inc., Lonwood Natural (NRMRL-RTP-460). Both
can be downloaded from the ETV Program website
(http://www.epa.gOv/etv/este.html#mrbmgw).
Original signed by
Sally Gutierrez
NRMRL Laboratory Director
Office of Research and Development
United States Environmental Protection Agency
Original signed by
Karin Foarde
Microbiology Department Director
Research Triangle Institute
NOTICE: ETV verifications are based on an evaluation of technology performance under specific, predetermined
criteria and the appropriate quality assurance procedures. EPA and RTI make no expressed or implied warranties as
to the performance of the technology and do not certify that a technology will always operate as verified. The end
user is solely responsible for complying with any and all applicable federal, state, and local requirements. Mention
of commercial product names does not imply endorsement.
EPA REVIEW NOTICE This report has been peer and administratively reviewed by the U.S. Enviromnental
Protection Agency, and approved for publication. Mention of trade names or commercial products does not
constitute endorsement or recommendation for use.
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