United States

Environmental Protection 

Agency

Office of Chemical Safety

and Pollution Prevention

(7101)

EPA 730-C-11-003

May 30, 2012

Product Performance

Test Guidelines

OCSPP 810.2500:

Air Sanitizers –

Efficacy Data

Recommendations
NOTICE

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

Environmental Protection Agency’s Office of Chemical Safety and Pollution Prevention 

(OCSPP) 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 Act (FFDCA) (21 U.S.C. 

346a). Prior to April 22, 2010, OCSPP was known as the Office of Prevention, Pesticides 

and Toxic Substances (OPPTS). To distinguish these guidelines from guidelines issued by 

other organizations, the numbering convention adopted in 1994 specifically included OPPTS 

as part of the guideline’s number. Any test guidelines developed after April 22, 2010 will 

use the new acronym (OCSPP) in their title.

The OCSPP 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 the 

test, 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. 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. At places 

in this guidance, the Agency uses the word “should.” In this guidance, the use of “should” 

with regard to an action means that the action is recommended rather than mandatory. The 

procedures 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 recommendations described in 

these guidelines, and the Agency will assess them for appropriateness on a case-by-case 

basis. 

For additional information about these test guidelines and to access these guidelines 

electronically, please go to http://www.epa.gov/ocspp and select “Test Methods & 

Guidelines.” 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-OPPT-2009-0576.
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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) (7 U.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 sanitization of air. 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] A statement that the 

study was conducted in accordance with this part; [b] A statement describing in detail all 

differences between the practices used in the study and those required by this part; or 

[c] A statement that the person was not a sponsor of the study, did not conduct the 

study, and does not know whether the study was conducted in accordance with 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 

applicant may have generated in support of the application. 
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The test protocol should address the following areas: description of aerosol 

generator and generation; description of medium used for generation; description of 

aerosol sampler including air volume sampled; volume of the chamber; particle size; 

aerosol dispersion of test material (vapor or particle dispersion; 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. 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 or space 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). 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 3 log10 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;

(viii) Statement of 40 CFR Part 160 Good Laboratory Practice compliance and 

Quality Assurance Statement;
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(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;

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., 
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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) 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.
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(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).