United States 

Environmental Protection 

Agency 

Office of Chemical Safety 

and Pollution Prevention 

(7510P) 

EPA 712-C-07-074 

Draft: May 11, 2011 

Product Performance 

Test Guidelines 

 OCSPP 810.2500: 

Air Sanitizers—Efficacy 

Data Recommendations 
2

NOTICE 

 This guideline is one of a series of test guidelines established by the Office of 

Chemical Safety and Pollution Prevention (OCSPP) (formerly the Office of Prevention, 

Pesticides and Toxic Substances (OPPTS) prior to April 22, 2010), United States 

Environmental Protection Agency for use in testing pesticides and chemical 

substances to develop data for submission to the Agency under the Toxic Substances 

Control Act (TSCA) (15 U.S.C. 2601, et seq.), the Federal Insecticide, Fungicide, and 

Rodenticide Act (FIFRA) (7 U.S.C. 136, et seq.), and section 408 of the Federal Food, 

Drug, and Cosmetic (FFDCA) (21 U.S.C. 346a), referred to hereinafter as the 

harmonized test guidelines. 

 The harmonized test guidelines serve as a compendium of accepted scientific 

methodologies and protocols that are intended to provide data to inform regulatory 

decisions under TSCA, FIFRA, and/or FFDCA. This document provides guidance for 

conducting appropriate tests, and is also used by EPA, the public, and the companies 

that are subject to data submission requirements under TSCA, FIFRA and/or the 

FFDCA. . At places in this guidance, the Agency uses the word “should.” In this 

guidance, use of “should” with regard to an action means that the action is 

recommended rather than mandatory. As a guidance document, these guidelines are 

not binding on either EPA or any outside parties, and the EPA may depart from the 

guidelines where circumstances warrant and without prior notice. The methods 

contained in this guideline are strongly recommended for generating the data that are 

the subject of the guideline, but EPA recognizes that departures may be appropriate in 

specific situations. You may propose alternatives to the methods recommended in 

these guidelines, with your supporting rationale. The Agency will assess such 

proposals on a case-by-case basis. 

 For additional information about OCSPP harmonized test guidelines and to 

access the guidelines electronically, please go to http://www.epa.gov/ocspp and select 

“Test Methods & Guidelines” on the left side navigation menu. You may also access 

the guidelines in http://www.regulations.gov grouped by Series under Docket ID #s: 

EPA-HQ-OPPT-2009-0150 through EPA-HQ-OPPT-2009-0159, and EPA-HQ-OPPT2009-0576. 

DRAFT DOCUMENT DISCLAIMER: This draft document is distributed solely for 

the purpose of external review. It has not been formally disseminated by the EPA 

and should not be construed to represent any Agency determination or policy. The 

information correction process under the Agency’s Information Quality Guidelines 

does not apply until this document is formally disseminated by the EPA in its final 

form. This draft document should only be cited or quoted in the context of 

providing comments. 
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OCSPP 810.2500: Air Sanitizers – efficacy data recommendations. 

 

 

 (a) Scope. 

 (1) Applicability. This guideline describes test methods that EPA believes will 

generally satisfy testing requirements of the Federal Insecticide, Fungicide, and 

Rodenticide Act (FIFRA) (7U.S.C. 136, et seq.) and the Federal Food, Drug, and 

Cosmetic Act (FFDCA) (21 U.S.C. 346a). It addresses testing to demonstrate the 

effectiveness of antimicrobial pesticides bearing label claims for the treatment of air to 

temporarily reduce the number of airborne bacteria. 

(2) Background. The source materials used in developing this OCSPP test 

guideline are OPP guidelines 91-5 Air sanitizers and 91-30 Acceptable methods 

(Pesticide Assessment Guidelines, Subdivision G, Product Performance, EPA report 

540/9-82-026, October 1982). 

 (b) Purpose. This guideline addresses efficacy testing for antimicrobial 

pesticides intended to be used for the treatment of air to temporarily reduce the 

number of airborne bacteria. 

 (c) General considerations. –(1) This guideline recommends methods for use 

in tests to be conducted to address the data requirements for pesticide registration. 

Good Laboratory Practice Standards (GLP) as defined in 40 CFR Part 160 apply to 

studies to support disinfection and sanitization of textiles. According to 40 CFR 

§160.17: “EPA may refuse to consider reliable for purposes of supporting an 

application for a research or marketing permit any data from a study which was not 

conducted in accordance with this part.” 40 CFR §160.12(b) requires with any 

submitted research data“[a] statement describing in detail all differences between the 

practices used in the study and those required by this part.” Refer to OCSPP Test 

Guideline 810.2000 for general testing recommendations prior to initiating tests. 

 (d) Air sanitizers. This section addresses efficacy testing for antimicrobial 

products intended to be used to temporarily reduce the number of airborne bacteria. 

 (1) Test procedure.—(A) There is no standard test method for evaluating the 

effectiveness of antimicrobial pesticides to temporarily reduce airborne bacteria. A list 

of references containing information and guidance on the testing of products intended 

for sanitizing the air of enclosed spaces is provided at paragraph (f) of this guideline. 

Applicants should contact the Agency and schedule a pre-registration meeting to 

discuss protocol development. Prior to the meeting, the applicant should provide an 

agenda of items for discussion, a draft label with the claims to be supported by the 

proposed protocol, and any research and development product performance data the 
4

applicant may have generated in support of the application. Quantitative 

microbiological assays should be performed to demonstrate the level of reduction of 

viable microorganisms achieved with the product, used as directed, in a sealed 

enclosure at least the size of a typical room (e.g., 10ft x 10ft x 8ft). The primary test 

bacteria for products intended for use in residential, commercial and industrial settings 

should be Staphylococcus aureus (American Type Culture Collection (ATCC) 6538) 

and Klebsiella pneumoniae (ATCC 4352). If the product is intended for use in 

hospitals or medical environments, the product should also be evaluated for efficacy 

against Pseudomonas aeruginosa (ATCC 15442). 

 In addition, the test protocol should address the following areas: method used 

to ensure sterility of the test enclosure prior to conducting the microbiological assays; 

the procedure for introducing microorganisms into the air of the chamber, as well as 

the sampling and recovery procedures employed; the environmental conditions in the 

room or chamber, such as temperature and relative humidity; and the concentration of 

chemical and contact time needed to demonstrate effectiveness. The settle rates in 

the air of the test enclosure should be measured for each of the recommended test 

bacteria. The study design should also include a parallel, untreated control. Raw 

data, as well as any statistical or graphical interpretation of the results, should be 

included in the study report. 

 (ii) Evaluation of sanitizer success. The test results should demonstrate a 

viable bacterial count reduction of ≥99.9 percent (a 3log10 reduction) over the parallel 

untreated control, after correcting for settling rates, in the air of the test enclosure with 

each of the required test bacteria within 5 minutes. 

(e) Data collection and reporting—(1) General. To assist in the proper 

review and evaluation of product performance, complete descriptions of the test 

employed and the results obtained should be submitted to the Agency. All test reports 

should include, at the least, the following information:

(i) Study title; 

(ii) Product Identity; 

(iii) Guideline number/Data Requirement; 

(iv) Identification of the testing laboratory or organization; 

(v) Location where the test was performed; 

(vi) Name(s) of the person(s) responsible for the test; 

(vii) Statement of Confidentiality Claims; 
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(viii) Statement of 40 CFR Part 160 Good Laboratory Practice compliance and 

Quality Assurance Statement; 

(ix) Purpose of the study; 

(x) Date and time of the start and end of the test; 

(xi) Test employed and any modifications (e.g., organic soil, hard water, etc), 

when using standard tests (e.g., AOAC, ASTM, etc) all deviations to the test methods 

should be reported; 

(xii) Test microorganisms employed, including identification of the specific 

strain (ATCC or other); 

(xiii) Description of the test substance, including the percent of active 

ingredient; 

(xiv) Concentration or dilution of the product tested and how prepared; 

(xv) Number of samples, batches and replicates tested; 

(xvi) Manufacture date of each product batch; 

(xvii) Identification of all material or procedural options employed, where such 

choice is provided for or recommended in the test method selected (e.g., growth 

media, drying time for inoculated carriers, neutralization confirmation and/or subculture 

media, secondary subculturing); 

(xviii) Test exposure conditions (e.g., contact time, temperature, and relative 

humidity); 

(xix) Complete reports of results obtained for each replication; 

(xx) Any control data essential to establish the validity of the test. 

(xxi) Carrier counts; 

(xxii) Statistical treatment of the data; 

(xxiii) Conclusions; 

(xxiv) References; 

(xxv) Appendices, including study protocol and all raw data reports associated 

with the conduct of the study; 
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The applicant is encouraged to use the EPA’s standard efficacy report format, 

which may be found at http://www.epa.gov/oppad001/efficacystudystandards.htm.

(2) Data for modifications of recommended methods. Where 

recommended methods are modified for testing conducted to support specific claims 

and/or use patterns for a product, the protocol, identifying and describing each 

modification (e.g., organic load, hard water), should be provided with the study report. 

The applicant is encouraged to submit the proposed modification to the Agency for 

review and evaluation prior to initiation of the test. 

(3) Data for other methods. When recommended methods, or modifications 

thereto, are not employed to develop efficacy data (such as actual in-use or many 

kinds of simulated-use testing), complete testing protocols should be submitted with 

the test reports. All materials and procedures employed in testing should be described 

in a manner consistent with original research reports published in technical or scientific 

journals. Where references to published reports or papers are made, copies or 

reprints of such references should be provided with the test reports. The applicant 

should submit the proposed testing protocols for in-use or simulated-use studies (with 

a proposed label to show the claims to be supported by the protocol) to the Agency for 

review and evaluation prior to initiation of the test. 

 

 (f) References. The references in this paragraph may be consulted for additional background information. 

(1) Brockmann, J.E. 1993. Sampling and Transport of Aerosols. In: Aerosol 

Measurement: Principles, Techniques and Applications, pp. 77 – 111. Eds. 

Willeke, K and Baron, P.A. Wiley New York, NY. 

(2) Buttner, M. P., Willeke, K., Grinshpun S. A. 1997. Sampling and Analysis of 

Airborne Microorganisms. In: Manual of Environmental Microbiology, pp 629 – 

640. Eds. Hurst, C.J., Knudsen, G. R., McInerney, M.J., et al., American 

Society for Microbiology, Washington, D. C. 

(3) Clark, S. and Scarpino, P. 1995. Disinfection of Microbial Aerosols. Chap 

7. In: Modeling Disease Transmission and its Prevention by Disinfection, 

Hurst, C.M., Cambridge University Press. 

(4) Cox, C.S. and Wathes, C.M. (Eds). 1995 Bioaerosols Handbook. Lewis 

Publications, Boca Raton, Fl. 

(5) Dimmick, R.L. and A.B. Akers (Eds.). An Introduction to Experimental 

Aerobiology. Wiley-Interscience, NY (1969). 

(6) Greene, V.W. et al. Microbiological contamination of hospital air. 

I. Quantitative studies. Applied Microbiology 10:561-566 (1962). 
7

(7) Gregory, P.H. and J.L. Monteith (Eds.). Airborne Microbes. Cambridge 

Univ. Press, London (1967). 

(8) Hirch, A., Lidwell, O.M., and Williams, R.E. 1956. The bacterial flora of the 

air of occupied rooms. J Hgg (Lond) Dec. 54 (4): 512 – 523. 

(9) Lester, W., Jr. et al. Factors of importance in the use of triethylene glycol 

vapor for aerial disinfection. American Journal of Public Health 40:813-820 

(1950). 

(10) Lester, W., Jr. et al. Bactericidal effects of propylene and triethylene 

glycol vapor on Escherichia coli. Science 115:379-382 (1952). 

(11) McGray, R.J. A test method for the evaluation of air sanitization. 

Processing Chemical Specialties Manufacturers Association pp.106-111 (1970). 

(12) McWall, R.E. 1975. Practical methods of reducing airborne contaminants 

in interior spaces. Arch Environ Health 30 (11): 552 – 558. 

(13) Puck, T.T., et al. The bactericidal action of propylene glycol vapor on 

microorganisms suspended in air. II. The influence of various factors on the 

activity of the vapor. Journal of Experimental Medicine 78:387-406 (1943). 

(14) Puck, T.T. The mechanisms of aerial disinfection by glycols and other 

chemical agents. II. An analysis of the factors governing the efficiency of 

chemical disinfection of the air. Journal of Experimental Medicine 85:729-757 

(1947). 

(15) Reponen, T., Willeke, K., Ulevicius, V., et al. 1997. Techniques for 

Dispersion of Microorganisms into air. Aerosol Sci. Technol. 27: 405- 421 

(16) Spendlove, J.C. and Fannin, K.F. 1983. Source, Significance and Control 

of Indoor Microbial Aerosols: Human Health Aspects. Public Health Rep. 98 

(3) May – June: 229 – 244. 

(17) Stuart, L.S. and J.L. Friedl. Testing aerosol products for germicidal and 

sanitizing activity. Processing Chemical Specialties Manufacturing Association, 

pp.93-97 (1955). 

(18) Wolf, H.W., et al. Sampling microbiological aerosols. Public Health 

Service Publication No. 686, Washington, DC (1959).