Document ID: EPA-HQ-OPPT-2009-0150-0026
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2013-03-13T04:00Z

United States
Environmental Protection Agency
   Office of Chemical Safety
   and Pollution Prevention
   (7101)
EPA 730-C-11-003
December 21, 2012

Product Performance
Test Guidelines

OCSPP 810.2600:

Disinfectants and Sanitizers for Use in Water  -  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" 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.
  

OCSPP 810.2600:  Disinfectants and sanitizers for treating water  -  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 disinfection or sanitization 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 systems, namely drinking water and swimming pools, as disinfectants or sanitizers. 
      
	  (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 of disinfectants for water. 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." Note:  The Association of Official Analytical Chemists (AOAC) recommended tests are designed to be conducted 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, obtain written approval from the Agency and document such deviations in the study reports submitted to the Agency. The Agency may consult with the AOAC prior to accepting modifications to their standardized methods. Refer to OCSPP Test Guideline 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 public water systems. Evidence should be submitted that the chemical intended for use as a drinking water disinfectant is in compliance with the EPA's Office of Drinking Water's regulations.

		(2)  Emergency water supplies. This section addresses efficacy testing for additives such as solutions, powders, or tablets intended for emergency disinfection of small quantities of drinking water of question - able potability (e.g., water obtained from lakes, streams, etc), by the 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 Water 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 triplicate.  The test organisms should be Raoultella terrigena  -  American Type Culture Collection (ATCC) 33257 (R. terrigena) (formerly designated as Klebsiella terrigena), Polio 1 (LSa) (ATCC VR-59), or Chat (ATCC VR-1562) Rotavirus (Wa or SA-11) (ATCC VR-2018 or VR-1565), Cryptosporidium parvum (C. parvum) and Giardia intestinalis (G. intestinalis) (formerly Giardia lamblia). Giardia muris may be tested in place of Giardia intestinalis.

	(ii) Evaluation of success -- (A)  Bacteria. A 6-log10 (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-log10 (99.99 percent) reduction of a mixed influent challenge of 1 x 10[7] organisms per liter of each of the following viruses: Po - liovirus (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-log10 reduction should be demonstrated. MS2 coliphage virus may also be used as a surrogate for Poliovirus and Rotavirus.
       

		(C)  Cysts (Protozoan).  A 3-log10 (99.9 percent) reduction of an influent challenge of 1 x 10[6] organisms per liter of C. parvum and G. intestinalis (or G. muris if tested) or a 3-log10 reduction of an influent challenge of 1 x 107 particles or spheres of 4 to 6 um diameter (as an option for units or components based on occlusion filtration, where disinfection or other active processes are not involved) 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 removal from water or other non-microbiological claims.  National Sanitation Foundation (NSF) Standard 42 (Aesthetic Effects) provides partial guidance for chemical removal.

		(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 alternative organism or laboratory procedure, or to respond to innovative treatment units. However, these alterations and/or additions should not decrease the level of testing or alter the intent of the protocol and should be submitted to the Antimicrobials Division for approval prior to initiation of testing. 

		(ii)  Evaluation of microbiological water purifier success -- (A)  Bacteria.  A 6-log10 (99.9999 percent) reduction of an influent chal - lenge 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-log10 (99.99 percent) reduction of a mixed influent challenge of 1 x 10[7] organisms per liter of each of the following viruses: Po - liovirus (LSa) (ATCC-VR-59) or Chat (ATCC 1562 and Rotavirus (Wa) (ATCC - -VR-2018) or SA11 (ATCC 1565) should be demonstrated by the testing. Virus types should be mixed in roughly equal 1 x 10[7]/L concentrations and a joint 4-log10 reduction should be demonstrated. MS2 coliphage virus may also be used as a surrogate for Poliovirus and Rotavirus.

	(C)   Cysts (Protozoan).  A 3-log10 (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) or Giardia muris (if tested) or a 3-log10 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 occlusion filtration, where disinfection or other active processes are not involved) should be demonstrated by the testing. Note: When using spheres as surrogates for oocysts, the charge of the surrogate particle, the shape of the particle, the size range of the particle, as well as the actual method of sample collection, are critical to the accurate and conservative evaluation of the reduction efficiency of size occlusion technologies.

	(D) Voluntary Optional Protocols. The Agency will consider protocols developed by the National Sanitation Foundation (NSF) International to support microbiological water purifier label claims. Specifically, NSF 53, NSF 55, NSF 58 and NSF 62 may be used where appropriate. NSF protocols P231 and P248 may also be used to generate data to support label claims as microbiological water purifiers. A pre-registration consultation is recommended prior to the use of any of these NSF standards to ensure the data will support the label claims.

	(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 concentrations necessary for  - disinfection of swimming pool water in practical applications: the number of swimmers/bathers in the pool; frequency of use; frequency with which water is changed; general weather conditions; and types and degree of or - ganic 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 test and confirmatory field test, is recommended. 

      (1)  Test procedure - Laboratory test.  The Agency recommends the use of the AOAC International Disinfectants (Water) for Swimming Pools (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 product that should be considered effective.

	(2)  Test procedure - Field test. The Agency recommends efficacy data to be derived from in-use tests under an experimental 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 disinfectant product during the field test should be conducted in the following manner: Water samples for bac - teriologic monitoring should be taken on opposite sides of the pool in the shallow area and as remote as possible from the inlets, preferably at the midpoint between the inlets.  A minimum of 144 samples (72 samples per pool) should be collected during the test period. Samples should 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 maximum bather load of the pool and the number of persons in the pool water at the time the samples are collected is at least 25 percent of the maximum 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 substances, metals, and water hardness) of the pool water at the sampling site should be determined 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 include (but are not limited to) the following information.

	(A)  The daily swimmer/bather load.

	(B)  The design of the pool, the recirculation 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 characteristics of the swimming pool water such as:  pH, nitrogenous substances, metals, and hardness.

	(E) The physical characteristics of the swimming pool water, includ - ing temperature and clarity, determined at least daily.

	(F)  Meteorological data, including air temperature, 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 field testing success.  The product, when used as recommended in swimming pool water, should demonstrate that not more than 15 percent of the sam - ples collected fail to meet the following bacterial indices (Ref. 3):

      (A) The standard plate count at 35°C not to exceed 200 colonies/1.0 mL.

      (B) The most probable number of coliform bacteria to be less than 2.2 organisms/100.0 mL.  When the membrane filter test is used, no more than 1.0 coliform organism/50 mL.

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

	(3) Voluntary Additional Test. The Agency recommends use of NSF/ANSI Standard 50. The test organisms should be Enterococcus faecium (ATCC 6569) and Pseudomonas aeruginosa (ATCC 27313).

	(i)  Evaluation of success. The product, when used as directed in swimming pool water should demonstrate a 3-log kill of the two target microorganisms, Enterococcus faecium and Pseudomonas aeruginosa.

 		(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 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;
 
 		(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.  When 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, 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.
 
 		(g)  References.  The references in this paragraph may be consulted for addi - tional background information.		
       
 			(1)	Guide Standard and Protocol for Testing Microbiological Water Purifiers.  United States Environmental Protection Agency, Registration Division, Office of Pesticide Programs and Criteria and Standards Division, Office of Drinking Water, April 1987.
 
 			(2) Official Methods of Analysis of the Association of Official Analyt - ical Chemists, Use-Dilution Methods, Chapter 6, Disinfectants; Current Edition.  Association of Official Analytical Chemists (AOAC), 2200 Wilson Boulevard Arlington, VA 22201.
 
 			(3)	Suggested Ordinance and Regulations Covering Public Swimming Pools.  Joint Committee on swimming Pools of the American Public Health Association, in Cooperation with the United States Public Health Service, 1964. The American Public Health Association, New York, NY.		
 
 			(4)	Standard Methods for the Examination of Water and Wastewater, Current Edition, Standard Methods Online:  http://www.standardmethods.org.	
 
 	     (5)  NSF/ANSI - 50 Equipment for Swimming Pools, Spas, Hot Tubs and other Recreational Water Facilities. NSF International 789 Dixboro Rd. POB 130140, Ann Arbor, MI 48113-0140.
 
 	    	(6) NSF/ANSI  -  53 Drinking Water Treatment Units  -  Health Effects. NSF International 789 Dixboro Rd. POB 130140, Ann Arbor, MI 48113-0140.