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Identifying Environmental Variables Important to the Testing and Assessment of Antimicrobial Treated Consumer Products.

Donald P. Satchell Ph.D. (Situ Biosciences LLC, Evanston, IL)

Background - Numerous factors are leading to the increased use and acceptance of antimicrobial products into consumer goods. Environmental sustainability initiatives are pushing for longer use or improved durability of finished goods and in development of new materials incorporating sustainable raw materials. Antimicrobial additives are a critical component to these initiatives, particularly in the use of recycled cellulose based materials which are used across a broad range of industries including Building Materials, Textiles, Paper, and Personal Care Products. Technologies for developing new formulations using antimicrobials rely on existing methods for demonstrating a level of performance. But specific factors affecting in-use antimicrobial performance remains somewhat misunderstood or unknown. As part of an initiative for developing practical methods for commercial and consumer product antimicrobial performance, a survey was made of environmental variables and their affects on test performance in two industry standardized ASTM methods. ASTM 3273 and ASTM 5590 are well accepted methods for the demonstration of antimicrobial performance in products from paper, to coatings such as adhesives, paints, and textiles. The hypothesis of the survey is to demonstrate that specific environmental exposures can greatly affect the antimicrobial performance of a given product and that these environmental variables can be appropriately incorporated into a the test strategy to improve the setting for the testing of the products, helping to better match the antimicrobials used in a given product with the real-world use of the finished goods. Although there are a multitude of potential variables, the environmental variables selected for the study were those hypothesized to result in the most significant physical or chemical affects on both the test material and/or antimicrobial additive. For coatings used in a building environment, particularly one potentially exposed to high humid conditions, the key environmental variables selected were temperature (high, low) and temperature cycling; moisture (via condensation) and age. The gross model application being mimicked is exposure of interior wall space, or enclosure such as a crawl space, attic space, inner wall space, or bath environment. Methods A minimum of 5 replicate samples were treated for each conditions using kiln dried white pine wood planks 1.25 X 5 x 0.125 (ASTM D3273) or 1.25x1.5x 0.125 (ASTM D5590) were selected as the control substrate for coating trials. Wood will naturally grow fungi when presented in an appropriate environment. Kiln dried wood will have a lower propensity for fungal growth but the samples were also autoclaved at 121C for 50 minutes, to further control the natural variability of fungal spores present in the native material. Coating material was acquired from manufacturers. The study protocol, evaluates three separate coating formulations, ranging from low, medium and high solids coating products. The solids content demonstrates a range if quality or cost of the coating product and is also generally representative of coating durability.

Environmental chambers utilizing computer controlled temperature and humidity were used as incubation chambers for the treated samples. Moisture Condensation: Treated samples were first chilled to 0C, and then placed in a 32C humidified chamber for 24 hours. The samples were cycled 60 times. Condensation was measured from ten samples as weight gain for 10 cycles to obtain an average value of water condensation. Temperature Cycling: In order to mimic relative extremes in temperature exposure, treated samples were temperature cycled at 12 hour intervals between 0C and 45C over a 90 day duration. Fungal Growth Test Methods: Two standardized ASTM test methods were used to determine the prevalence of fungal growth on the samples. ASTM D3273, is a native product chamber test. The essence of which provides appropriate conditions for fungal growth in an enclosed environment with high levels of fungal spore exposure but no additional constituents other than the sample material are present. ASTM D5590 is an enhanced test method in which fungal growth is supplemented by the presence of a complex media to provide positive fungal growth in the absence of antimicrobial agents. Both test methods use a mix sample of common fungi species found in a housing environment (Aspergillus niger ATCC 6275, Penicillium funiculosum ATCC 11797, Aureobasidium pullulans ATCC 9348). ASTM D3273 inoculation is based on fungal spore growth from a dirt matrix positioned below hung samples and ASTM D5590 inoculation is provided by direct misting of a fungal spore solution onto samples placed in contact with a potato dextrose agar plate. Incubation for each method traditionally lasts for 28 days, but longer incubation periods can be used to accommodate comparisons of higher durability samples. The incubation temperature is 28 to 32C.

Results The objective of the study is to highlight relevant test variables that can be appropriately incorporated into to a product validation scheme to better assess antimicrobial products formulations and their constituent components for a specific product application. In our test program, samples were exposed to environmental variables identified as important to interior building environments. These environments were selected due to the significant challenges in the building and construction industry for mitigation of fungal growth, and the relative lack of information on best practices for application development and use of appropriate antimicrobial chemistries. Temperature has several effects on building surfaces. In addition to being a requisite for microbial growth, fluctuations in temperature stress coating surfaces potentially leading to cracking of a coating barrier and moisture condensation. Improved building practices are critical to mitigating these extremes but new composite materials based on wood pulp are also increasing these challenges due to their increased propensity for growing fungi. The results from our testing show that the most significant environmental variable present in this study is the presence of moisture. Sample cycled through condensation demonstrated both more vigorous and rapid growth of fungi under both test conditions. This result is significant as it indicates that the additive used in these samples is readily soluble or degraded in moisture. Additional studies evaluating aqueous solubility and comparisons with identical formulation will better illustrate

the need for improved fit for purpose formulation of the antimicrobial for this type of application. Temperature cycling demonstrated a great impact on coating containing low solids. Very like due to the lack of resiliency of the coating and subsequent access of fungal spores to the wood matrix. Images show significant cracking, with which is also dramatically represented in the water condensation test samples. Accelerated aging, demonstrated statistically significant decrease of approximately 20% in performance as measure by ASTM D3273 and a much greater decrease in ASTM 5590. This is likely due to the loss of volatile components in the test materials. Temperature aging is difficult in these sample types due to the difference in volatility of some additives. In this study we attempted to limit serious loss by limiting the temperature range used. Ideally a temperature dependent comparison should be performed to better understand the effect of accelerated aging in assessing antimicrobial formulation performance.

Conclusion In this study we show a clear correlation between environmental variable durability challenges and the resulting performance of an antifungal coating surface. Statistically, assessing several pragmatic factors, the strongest factor affecting performance is the presence of moisture. Leaching or degradation due to the presence of the moisture is causing the loss of the antimicrobial performance. Importantly, reproducible and comparable results can be clearly demonstrated for commercial and consumer product development by incorporating well known strategies. These strategies can then be used in developing more appropriate and fit for purpose formulation, while meeting environmental sustainability, safety, and cost targets for antimicrobial additives in consumer products.