United States

Environmental Protection Agency Office of Chemical Safety

and Pollution Prevention (7510P) EPA 712-C-07-074 May 11, 2011



# Product Performance
Test Guidelines

OCSPP 810.2600:
Disinfectants for Use in Water—Efficacy Data

Recommendations2 NOTICE

This guideline is one of a series of test guidelines established by the Office of Chemical Safety and Pollution Prevent ion (OCSPP) (formerly the Office of Prevention, Pesticides and Toxic Subst ances (OPPTS) prior to April 22, 2010), United States Environmental Protection Ag ency for use in testing pesticides and chemical substances to develop data fo r 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 t he harmonized test guidelines.

The harmonized test guidelines se rve as a compendium of accepted scientific methodologies and protocols that are intended to provide data to inform regulatory decisions under TSCA, FIFR A, and/or FFDCA. This document provides guidance for conducting appropriate tests, and is also used by EPA, the public, and the companies that are subj ect 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 ra ther 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 ci rcumstances warrant and without prior notice. The methods cont ained in this guideline are strongly recommended for generating the data that ar e the subject of the guideline, but EPA recognizes that departures may be appropr iate 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-bycase 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-OPPT-2009-0576.

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 repr esent 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.3 OCSPP 810.2600: Disinfectants for tr eating water – efficacy data

recommendations.

(a) Scope.—

(1) Applicability . This guideline describes test methods that EPA believes will generally satisfy testing r equirements 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 bear ing label claims for the disinfection of

water.

(2) Background . The source materials used in developing this OCSPP test guideline are OPP guidelines 91-8 Products for treating water systems 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 in water syst ems, namely drinking water and swimming pools, as disinfectants.

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

tests to be conducted to addre ss the data requirements for pes ticide registration. Good Laboratory Practice Standards (GLP) as defined in 40 CFR Part 160 apply to studies of disinfectants for water systems. Accordi ng 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 res earch data “[a] statement describing in detail all differences between the practices us ed in the study and those required by this part.” Note: The Association of Official Analytical Chemists (AOAC) recommended tests are designed to be conduct ed as written. For deviations (e.g., cultures grown with shaking instead of static, dilution of culture prior to drying on carriers) proposed to be used in the conduct of these tests, obtai n written approval fr om the Agency and document such deviations in the study repor ts submitted to the Agency. The Agency may consult with the AOAC prior to acc epting modifications to their standardized

methods. Refer to OCSPP Test Gu ideline 810.2000 for general testing recommendations prior to initiating tests.

(d) Drinking water for humans

— (1) Public water systems . Applicants should contact EPA’s Antimicrobials Division prior to conducting studies to support label claims for disinfection of publ ic water systems.

(2) Emergency water supplies . This section addresses efficacy testing for additives such as solutions, powders, or t ablets intended for emergency disinfection of small quantities of drinking water of questionabl e potability (e.g., water obtained from4 lakes, streams, etc), by t he general public, in the absence of bacteriological monitoring

facilities.

(i) Test procedure . The Agency recommends the use of The Guide Standard and Protocol for Testing Microbiological Wate r Purifiers (Ref. 1). Three samples of product, representing three different batches , one of which should be at least 60 days old, should be tested in triplicat e. The test organisms should be Raoultella terrigena – American Type Culture Co llection (ATCC) 33257 ( R. terrigena ) (formerly designated as Klebsiella terrigena ), Polio 1 (LSa) (ATCC VR-59), Rotavirus (Wa) (ATCC VR-2018), Cryptosporidium parvum ( C. parvum ) and Giardia intestinalis ( G. intestinalis ) (formerly

Giardia lamblia)

. Giardia muris may be tested in place of Giardia lamblia.

(ii) Evaluation of success

— (A) Bacteria. A 6-log 10 (99.9999 percent) reduction of an influent challenge of 1 x10 7 organisms per 100 mL of the bacterial challenge organism R. terrigena should be demonstrated by the testing.

(B) Viruses. A 4-log 10 (99.99 percent) reduction of a mixed influent challenge of

1 x 10 7 organisms per liter of each of the following viruses: Poliovirus (LSa) and

Rotavirus ( Wa) should be demonstrated by the testing. Virus types should be mixed in roughly equal 1 x 10 7 /L concentrations and a joint 4-log 10 reduction should be

demonstrated.

(C) Cysts (Protozoan). A 3-log 10 (99.9 percent) reduction of an influent challenge of 1 x 10 6 organisms per liter of C. parvum and G. intestinalis (and G. muris if tested) should be demonstrated by the testing.

(3) Water purifier units. This section addresses efficacy testing for products bearing label claims for microbiological water purification (removal of pathogenic microorganisms). This section does not apply to products only used to treat water in order to achieve specific chemical remova l from water or other non-microbiological claims. National Sanitation Foundation (NSF ) Standards 42 (Aesthetic Effects) and 53 (Health Effects) provide partial guidance for chemical removal and other claims testing.

(i) Test procedure. The Agency recommends the use of The Guide Standard and Protocol for Testing Microbiological Water Purifiers (Ref. 1). This is a general guide and in some cases may present only the minimum features and framework for the testing of microbiological water purifiers. Users of the Guide Standard and Protocol should include pre-testing of their units in a testing rig, including the sampling techniques to be used. In some cases, it may be appropriate to alter and/or add to the guide in order to meet specific operational problems, to use an al ternative organism or laboratory procedure, or to respond to innovative treatment units. However, these

alterations and/or additions should not decrease t he level of testing or alter the intent of the protocol and should be submi tted to the Antimicrobials Div ision for approval prior to initiation of testing.5 (ii) Evaluation of microbi ological water purifier success

— (A) Bacteria. A 6- log 10 (99.9999 percent) reduction of an influent challenge of 1 x 10 7 organisms per 100

mL of the bacterial challenge organism Raoultella terrigena (ATCC) 33257) should be demonstrated by the testing.

(B) Viruses. A 4-log 10 (99.99 percent) reduction of a mixed influent challenge of

1 x 10 7 organisms per liter of eac h of the following viruses: Poliovirus(LSa) (ATCC-VR59) and Rotavirus ( Wa) (ATCC-VR-2018) should be demons trated by the testing. Virus types should be mixed in roughly equal 1 x 10 7 /L concentrations and a joint 4-log 10 reduction should be demonstrated.

(C) Cysts (Protozoan). A 3-log 10 (99.9 percent) reduction of an influent challenge of 1 x 10 6 organisms per liter of Cryptosporidium parvum and Giardia

intestinalis (formerly Giardia lamblia ) and Giardia muris (if tested) or a 3-log 10 reduction of an influent challenge of 1 x 10 7 particles or spheres of 4 to 6 um diameter (as an option for units or components based on occlusi on filtration, where disinfection or other active processes are not involved) should be demonstrated by the testing.

(e) Swimming pool, spa, and hot tub water . This section addresses efficacy testing for disinfectants bearing label claims for use in swimming pools, spas, and hot

tubs. Numerous factors influence the concent rations necessary for disinfection of swimming pool water in practical applications : the number of swimmers/bathers in the pool, frequency of use, frequency with whic h water is changed; general weather conditions; and types and degree of organic contamination of the water by the swimmers/bathers themselves (e.g., suntan lotion and oils) and by various debris. Therefore, a two-phased study, consisting of a laboratory te st and confirmatory field test, is recommended.

(1) Test procedure - Laboratory test . The Agency recommends the use of the

AOAC International Disinfec tants (Water) for Swimming P ools (Ref. 2). The test organisms should be Escherichia coli ( E.coli ) (ATCC 11229) and Enterococcus faecium (E. faecium ) (ATCC 6569) (formerly designated as Streptococcus faecalis ).

(i) Evaluation of success. The lowest concentration of the test substance providing results equivalent to those of the sodium hypochlorite reference standard is the lowest concentration of the produc t that can be considered effective.

(2) Test procedure - Field test. The Agency recommends efficacy data to be derived from in-use tests under an experim ental use permit (EUP) in at least two swimming pools, using the following components.

(i) Bacteriologic monitoring. Bacteriologic monitoring of swimming pool water following treatment with disinfec tant product during the field test should be conducted in the following manner: Water samples for bacteriologic monitoring should be taken on opposite sides of the pool in t he shallow area and as remote as possible from the inlets, preferably at the midpoint bet ween the inlets. A minimum of 144 samples should be6 collected during the test period. Samples s hould be taken just below the surface of the water and preferably at such times when the number of persons using the pool during the preceding hour has been at least equal to 50 percent of the maxi mum bather load of the pool and the number of per sons in the pool water at the time the samples are collected is at least 25 percent of the ma ximum bather load (as defined in Suggested Ordinance and Regulations Covering Public Swimming Pools, APHA Joint Committee on Swimming Pools and Bathing Places) of the pool (Ref. 3). Pertinent chemical characteristics (e.g., pH, nitrogenous subst ances, metals, and water hardness) of the pool water at the sampling site should be dete rmined at the time of sampling. (Ref. 4)

(ii) Field test. Field tests should be conducted for an entire swimming season (4 to 12 months). Reports should incl ude (but are not limited to) the following

information.

(A) The daily swimmer/bather load.

(B) The design of the pool, the reci rculation and filter system, and water capacity.

(C) The amount and identification of all chemicals added daily to the swimming pool water (including the time, site, and method).

(D) The range of chemical characteristi cs of the swimming pool water such as: pH, nitrogenous substances, metals, and hardness.

(E) The physical characteristics of the swimming pool water, including temperature and clarity, det ermined at least daily.

(F) Meteorological data, including air te mperature, rainfall, and number of hours of sunlight (determined daily) for outdoor pools.

(G) Bacteriologic monitoring, conducted daily, in accordance with the suggested Ordinance and Regulations Covering Public Swimming Pools of the American Public Health Association. (Ref. 3)

(H) The concentration of the antimicrobial agent in the swimming pool monitored daily at the same time intervals that the bacteriological assay samples are obtained.

(I) The method that the product user will employ for monitoring the level (concentration in ppm) of antimicrobial agent contained in the pool water.

(iii) Evaluation of fi eld testing success. The product, when used as recommended in swimming pool water, should demonstrate that not more than 15 percent of the samples collected fail to meet the following bacter ial indices (Ref. 3):

(A) The standard plate count at 35°C not to exceed 200 colonies/1.0 mL.7 (B) The most probable number of colif orm bacteria to be less than 2.2 organisms/100.0 mL. When the me mbrane filter test is used, no more than 1.0 coliform organism/50 mL.

(C) The most probable number of enteroc occal organisms to be less than 2.2 organisms/100 mL. When the membrane filt er test is used, no more than 1.0 enterococcal organism/50 mL.

(f) 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 A gency. 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 testi ng laboratory or organization;

(v) Location where the test was performed;

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

(vii) Statement of Conf identiality Claims;

(viii) Statement of 40 CFR Pa rt 160 Good Laboratory Prac tice compliance and Quality Assurance Statement;

(ix) Purpose of the study;

(x) Date and time of the st art 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 deviat ions 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, includ ing the percent of active ingredient;

(xiv) Concentration or dilution of the product tested and how prepared;8 (xv) Number of samples, batches and replicates tested;

(xvi) Manufacture date of each product batch;

(xvii) Identification of all material or pr ocedural options employed, where such choice is provided for or recommended in the te st method selected (e .g., growth media, drying time for inoculated carriers, neutralizati on confirmation and/or subculture media, secondary subculturing);

(xviii) Test exposure conditions (e.g., contac t time, temperature, and relative humidity);

(xix) Complete reports of result s obtained for each replication;

(xx) Any control data essential to est ablish the validit y of the test;

(xxi) Carrier counts;

(xxii) Statistical treatm ent 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. When recommended methods are modified for testing conducted to support specific claims and/or use patterns for a product, the protocol, identifyi ng and describing each modification, should be provided with the study report. The appl icant is encouraged to submit the proposed modification to the Agency fo r 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 testi ng protocols should be s ubmitted with the test reports. All materials and procedures em ployed 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 ar e made, copies or reprints of such references should be provided with the test reports. The applicant should submit the proposed testing pr otocols for in-use or simu lated-use studies (with a proposed label to show the claims to be sup ported by the protocol) to the Agency for review and evaluation prior to initiation of the test.9 (g) References. The references in this paragr aph may be consulted for additional background information.

(1) Guide Standard and Protocol for Testi ng Microbiological Water Purifiers

. United States Environmental Protection Ag ency, Registration Division, Office of Pesticide Programs and Criter ia and Standards Division, Office of Drinking Water, April 1987.

(2) Official Methods of Analysis of the Asso ciation of Official Analytical Chemists

, Use-Dilution Methods, Chapter 6, Disinfect ants; Current Edition. Association of Official Analytical Chemis ts (AOAC), 2200 Wilson Boul evard Arlington, VA 22201.

(3) Suggested Ordinance and Regulations Covering Public Swimming Pools

. Joint Committee on swimming Pools of the Am erican Public Health Association, in Cooperation with the United St ates Public Health Service, 1964. The American
Public Health Association, New York, NY.

(4) Standard Methods for the Exami nation of Water and Wastewater , Current Edition, Standard Methods Online: http://www.standardmethods.org.