Document ID: EPA-HQ-OPPT-2009-0150-0015
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2012-03-14T04:00Z

United States
Environmental Protection
Agency

Office of Chemical Safety and Pollution Prevention
(7510P)
	EPA 712-C-07-091
                                                                	March 12, 2012
	Product Performance Test Guidelines
		
		OCSPP 810.2300:
            Sanitizers for Use on Hard Surfaces -- Efficacy Data Recommendations

                                       
                                       
                                       
                                       

                                        
                                     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-OPPT-2009-0576.
  

OCSPP 810.2300:  Sanitizers for use on hard surfaces - 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 effectiveness of antimicrobial pesticides bearing claims for use as sanitizers.  
      
      (2) Background. The source materials used in developing this OCSPP test guideline are OPP guidelines 91-2: Products for use on hard surfaces 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 on hard surfaces, namely sanitizers in a variety of product types (water-soluble powders, liquids, sprays, towelettes, etc.).    
      
      (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 on hard, non-porous surfaces.  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 Chemicals (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.
      
      (2)  Confirmatory testing. In certain situations an applicant may rely on previously submitted efficacy data to support an application or amendment for registration of a product and submit only confirmatory efficacy data on his own product to demonstrate his ability to produce an effective formulation. These situations are as outlined in paragraphs (c)(2)(i) and (c)(2)(ii) in this guideline.
      
      (i)  Duplicated Product Formulations. In this situation, the applicant manufactures a formulation which duplicates a product that is already registered with complete supporting efficacy data. The chemical composition, manufacturing procedure, label claims, and directions for use are identical in substance to those of the original registration, and specific references (Master Record ID Numbers [MRIDs]) to the supporting data developed for the original product are cited by the applicant.
      
      (ii)  Minor Formulation Change in a Registered Product. In this situation, the change in the formulation is relatively minor, e.g., a change of an inert ingredient. The label claims and directions for use are unchanged from those accepted for the registered formulation, and specific references (MRIDs) to the supporting data developed for the original formulation are cited by the applicant. If the only change in the formulation is the addition of a fragrance or dye, confirmatory data do not need to be submitted. However, when the product is an aerosol formulation, confirmatory data should be submitted for all formulation changes, including the addition of fragrances and dyes.  
      
      
      Table 1. Tests for basic claims described in this guideline. 
Level of Efficacy
Test Methods
Test Organisms
No. of Batches/Carriers
Evaluation of Success
Non-food Contact Sanitizer
Water soluble powders/liquids
 Spray products
ASTM E-1153 
Staphylococcus aureus (ATCC 6538) and Klebsiella pneumoniae (ATCC 4352) Enterobacter aerogenes (ATCC 13048) may be substituted for K. pneumoniae.

Three batches, one at least 60 days old.
99.9% reduction within 5 minutes. 

Towelettes
Reserved

Food Contact Surface Sanitizer
Halide Products
Water soluble powders/liquids
AOAC International Chlorine (Available) in Disinfectants Germicidal Equivalent Concentration

Salmonella enterica (formerly typhi) (ATCC 6539) or S. aureus (ATCC 6538)

Three batches, one at least 60 days old.
Test results should demonstrate product concentrations equivalent in activity to 50, 100, and 200 ppm of available chlorine.
Food Contact Surface Sanitizer
Non-Halide Products
Water soluble powders/liquids
AOAC International Germicidal and Detergent Sanitizing Action of Disinfectants

Escherichia coli (ATCC 11229) and S. aureus (ATCC 6538)
Three batches, one at least 60 days old.  
99.999% reduction in the number of each test microorganism within 30 seconds.
Food Contact Surface Sanitizer
Towelette
Towelettes

http://www.epa.gov/oppad001/towelette

Sanitizers for Urinal and Toilet Bowl Water and In-tank Sanitizers

Water soluble powders/liquids/tablets
Simulated-use study

Enterococcus faecium ATCC 6569 or Salmonella enterica (formerly choleraesuis) ATCC 10708
Three batches, one of which is at least 60 days old.
99.9% reduction over parallel control counts.
Residual Self-sanitizing  -  wet surfaces

Simulated-use study
Representative gram positive and gram negative organisms
Three batches, one at least 60 days old.
99.9% reduction over parallel control counts.
      
       (iii) The confirmatory data are to be developed from testing the applicant's own finished product. When the test methodology utilized in deriving the original supporting efficacy data were modified to include additional elements not specified in the recommended method, such as organic soil, hard water, longer or shorter contact time, etc., the confirmatory data should be produced under similarly modified conditions.
      
      (3)  Table 1 provides a quick reference guide to testing for basic claims described in this guideline. Consult the text for detailed testing descriptions.
      
      (d) Sanitizers for nonfood contact surfaces (water soluble powders, liquids, and spray products). These products, when used as directed, should reduce the number of test microorganisms on a treated surface over those of an untreated control surface. The following testing recommendations apply to products bearing label claims for effectiveness as sanitizers for inanimate hard surfaces other than those which come in contact with food or beverages (e.g., floors, walls, furnishings).
      
      (1) Test Procedures.  
      
      (i) The Agency recommends the test procedure in this paragraph:  The American Society for Testing and Materials (ASTM) Test Method for Efficacy of Sanitizers Recommended for Inanimate Non-Food Contact Surfaces (E1153) may be used (Ref. 1). Three product samples, representing three different batches, one of which should be >=60 days old, should be tested against each test bacterium on each representative test surface depending on the uses proposed on the label. For hard, porous surface label claims use unglazed ceramic tile for hard, nonporous surface label claims use stainless steel carrier or glass slide, using 5 test carriers and 3 control carriers. The test microorganisms are: (S. aureus) (ATCC 6538) and (K. pneumoniae) (ATCC 4352). E. aerogenes (ATCC 13048) may be substituted for K. pneumoniae. The ASTM method states "an average of at least 7.5 x 105 organisms must have survived the inoculated control squares for the test to be valid." 
      
      (ii) Evaluation of sanitizing success for nonfood contact surface sanitizers. The results should demonstrate a reduction of >=99.9% (a 3-log10 reduction) in the number of each test microorganism over the parallel control count within 5 minutes.
      
      (e)  Towelettes. (Reserved.)
      
      (f)  Sanitizers for Internal Toilet and Urinal Bowl Surfaces Above and Below the Water Line
      
      (1) Test Procedures. The Agency recommends the use of The American Society for Testing and Materials (ASTM) Test Method for Efficacy of Sanitizers Recommended for Inanimate Non-Food Contact Surfaces (E1153) may be used (Ref. 1). The contained bowl water (96 fl oz which represents traditional high volume toilets) should used to calculate the appropriate use dilution for testing. The water volume in low volume toilet bowls should be measured and used to calculate the appropriate dilution for testing.
      
      (2) Evaluation of sanitizing success for toilet and urinal bowl surface sanitizers. The results should demonstrate a reduction of >=99.9% (a 3-log reduction) in the number of each test microorganism over the parallel control count within 5 minutes.
      
      (g)  Sanitizing rinses (soluble powders and liquids) for previously cleaned food-contact surfaces. This section addresses efficacy testing for products with a label rec - ommendation for the treatment of previously cleaned, nonporous, food -  contact surfaces (e.g., eating and drinking utensils and food processing equipment) as a terminal sanitizing rinse.  
      
      (1) Halide chemical products. Sanitizing rinses formulated with iodophors, mixed halides, and chlorine-bearing chemicals.
      
      (i) Test procedure. The Agency recommends the AOAC International Chlorine (Available) in Disinfectants Germicidal Equivalent Concentration test (Ref. 2). Three samples, representing three different batches, one of which should be at least 60 days old, should be evaluated for efficacy against Salmonella enterica (S. enterica) (formerly Salmonella typhi) (ATCC 6539) or S. aureus (ATCC 6538). When claims are made for the effectiveness of the product in hard water, all data should be developed at the hard water tolerance claimed. 
      
      (ii) Evaluation of sanitizing success of halide formulations. Test results should demonstrate product concentrations equivalent in activity to 50, 100, or 200 ppm of available chlorine. The reference standard is sodium hypochlorite.
      
      (2) Confirmatory testing for halide chemical products
      
      (i) Test procedure. The Agency recommends the AOAC International Chlorine (Available) in Disinfectants Germicidal Equivalent Concentration test (Ref. 2). One sample should be evaluated for efficacy against S. enterica (ATCC 6539) or S. aureus (ATCC 6538). When claims are made for the effectiveness of the product in hard water, all data should be developed at the hard water tolerance claimed. 
      
      (ii) Evaluation of sanitizing success of halide formulations. Test results should demonstrate product concentrations equivalent in activity to 50, 100, or 200 ppm of available chlorine. The reference standard is sodium hypochlorite.
      
      (3)  Non-halide chemical products. Sanitizing rinses formulated with qua - ternary ammonium compounds, chlorinated trisodium phosphate and ani - onic detergent-acid formulations.
      
      (i) Test procedure. The Agency recommends the AOAC International Germicidal and Deter - gent Sanitizing Action of Disinfectants test (Ref. 3). Three samples, representing three different batches, one of which should be at least 60 days old, should be evaluated for effi - cacy against both Escherichia coli (E.coli) (ATCC 11229) and S. aureus (ATCC 6538). When claims are made for the effectiveness of the product in hard water, all data should be developed at the hard water tolerance claimed. 
      
      (ii) Evaluation of sanitizing success of non-halide formulations. Acceptable results should demonstrate a >=99.999% reduction in the number of each test microorganism within 30 seconds. The results should be reported according to the actual count and percentage reduction over the control. 
      
      (4) Confirmatory testing for non-halide products -- (i) Test procedure. The Agency recommends the AOAC International Germicidal and Deter - gent Sanitizing Action of Disinfectants test (Ref. 3). One sample should be evaluated for effi - cacy against both E. coli (ATCC 11229) and S. aureus (ATCC 6538). When claims are made for the effectiveness of the product in hard water, all data should be developed at the hard water tolerance claimed. 
      
      (ii) Evaluation of sanitizing success of non-halide formulations. Acceptable results should demonstrate a >=99.999% reduction in the number of each test microorganism within 30 seconds. The results should be reported according to the actual count and percentage reduction over the control.
      
      (h)  Towelette Sanitizers for Food Contact Surfaces. This section addresses efficacy testing for products with a label recommendation for the treatment of hard, non-porous surfaces which may come into contact with food. Food contact surface (FCS) towelettes are intended to be used to sanitize the following surfaces: hard non-porous tables, countertops (stainless steel, laminated, sealed ceramic) stove tops, interior and exterior surfaces of microwaves and refrigerators. FCS towelettes may not be used to sanitize the following food contact surfaces: utensils, glasses, food containers, dishes, cutting boards, cutting blocks, drain boards and food processing equipment. This list is not meant to be all-inclusive, but to serve as general guidance for the appropriate use of this type of antimicrobial pesticide. The Agency reserves the right to accept or deny use sites for food contact surface towelettes on a case-by-case basis.
      
      (1) Test Procedure. The Agency recommends the use of the Interim Guidance for Non-Residual Sanitization of Hard Inanimate Food Contact Surfaces Using Pre-Saturated Towelettes. This guidance may be found at:  http://www.epa.gov/oppad001/towelette.htm
      
      Three samples, representing three different batches, one of which is at least 60 days old, should be evaluated for efficacy against E. coli (ATCC 11229) and S. aureus (ATCC 6538). Based on the claims, a variety of surfaces may be treated with the product. Each of the different types of test surfaces claimed may be used in the efficacy testing of the product (i.e., glass, stainless steel, plastic, and ceramic). At a minimum, the applicant should test: 1) a stainless steel or glass surface, and 2) a plastic with a rough surface (i.e., plastic cutting boards). Inoculate the test surface with the challenge microorganisms. After inoculation, the test surface should be dried for 40 minutes in an incubator at 30 - 37EC. A "zero-time" bacterial numbers recovery test should be performed to demonstrate the efficiency of the recovery process, and should be be reported.
      
      The towelette removed from its container should be handled with sterile gloves. The inoculated surfaces should be tested by wiping the surfaces with the saturated towelette. One towelette should be used to wipe 5 carriers. The area of the towelette used for wiping should be rotated so as to expose a maximum amount of its surface in the course of wiping the contaminated test surface. After wiping the contaminated surface with the towelette, all remaining liquid should be expressed from the used towelette into an empty sterile container and subcultured separately. Parallel tests on the towelette (as well as expressed liquid from the used towelette) should be run with the active ingredients omitted in an identical manner to serve as the control.
      
      
      (2) Evaluation of towelette sanitizing success. The product should demonstrate a >=99.999% mean reduction in the number of test microorganisms (bacteria) within 30 seconds. The result should be reported according to the actual count and percentage reduction over the control.
      
      (i) Sanitizers for Toilet and Urinal Bowl Water. This section addresses efficacy testing for products with claims as sanitizers for toilet and urinal bowl water.
      
      (1) Test Procedure. A simulated-use study should be designed which incorporates all of the elements listed in paragraphs (i)(1)(i) through (i)(1)(iv) in this guideline.
      
      (i)  The product should be added to samples of the bowl water from three toilets or urinals, one toilet/urinal per lot, at the use concentration employing the recommended method of dispensing. Untreated control samples from the three toilets or urinals should also be included.
      
      (ii)  Whether the product is automatically metered, or dispensed in some other fashion, into the bowl water (or urinal trap), the consistent accuracy of the concentration dispensed and maintained should be docu - mented.
      
      (iii)  Inocula containing representative Gram-positive (e.g., Enterococcus faecium ATCC 6569) or Gram-nega - tive (e.g., Salmonella enterica ATCC 10708) test bacteria should be added to the treated and control samples of the bowl water from each of the toilets or urinals to provide a concentration of at least 10[4] colony-forming units per milliliter.
      
      (iv)  Microbial counts of the treated bowl water and the control bowl water should be conducted at a minimum of three exposure intervals, equating to the duration of the product, ( "x" number of flushes/day/ "x" weeks), in addition to a 0-time control.
      
      (2) Evaluation of Sanitizing Success. The reduction of each test microorganism should be at >=99.9% mean reduction over the 0-time control and the parallel un - treated inoculated control.
      
      (j) In-Tank Sanitizers. This section addresses efficacy testing for products which bear label claims for use as an automatically dispensed in-tank sanitizer.
      
      (1) Test Procedures. In-tank sanitizer products should be evaluated by a preliminary simulated-use test followed by a laboratory efficacy test. These tests should incorporate the elements in paragraphs (j)(1)(i) through (j)(1)(i)(B).
      
      (i) Preliminary simulated-use test. The use-life of the in-tank product should be documented for three product samples, each in a separate toilet under the conditions in paragraphs (j)(1)(i)(A) and (j)(1)(i)(B) of this guideline simulating actual usage. Testing conducted with a 6 gallons/flush toilet may be used to generate data for low flush toilets (3.5 gallons/flush).
      
      (A) Number of flushes (dispensation of the dosage) per day per X weeks (duration of effectiveness) with a non-chlorinated water supply at 25-30 C (the warm water temperature extreme in summer).
      
      (B) The bowl water should be analyzed at periodic intervals during the testing indicated in paragraph (j)(1)(i)(A) in this guideline to demonstrate the pH and concentration of the active in - gredients.
      
      (ii) Laboratory efficacy tests. Bacteriologic assays should be conducted on treated and untreated samples which are neutralized by standard procedures employing:
      
      (A) Samples of the residual bowl water from three toilets (one lot per toilet) (at the mini - mal use concentration) and corresponding untreated control samples from three toilets at 10-.15 C (the most stringent water temperature for dem - onstrating efficacy).
      
      (B)  Representative Gram-positive and Gram-negative bacteria (e.g., Enterococcus faecium ATCC 6569, and Salmonella enterica ATCC 10708) with an inoculum of at least 10[4]CFU/mL.
      
      (C)  A minimum of three exposure intervals, in addition to a 0-time control.
      
      (iii) Evaluation of in-tank sanitizing success. The reduction of each test microorganism should be >=99.9% mean reduction over the 0-time control and the parallel untreated inoculated control.
      
      (k) Residual self-sanitizing activity of dried chemical residues on hard-inanimate surfaces - wet surfaces. This section addresses efficacy testing for products which bear label claims to provide residual self-sanitizing activity (e.g., significant reduction in numbers of infectious microorganisms which may be present or subsequently deposited) on treated surfaces that are likely to become and remain wet under normal conditions of use.
      
      (1) Test procedure. Residual self-sanitizing products for use on hard, non-porous, inanimate surfaces should be evaluated for efficacy using a controlled in-use study or simulated in-use study. The design of the study should be done in consultation with the Agency and should include the basic elements set forth in paragraphs (k)(1)(i) through (k)(1)(vii) of this guideline. 
      
      (i) The test microorganisms employed in the study should be pathogens that are likely to be encountered in the environment in which the product is to be used.
      
      (ii) The starting inocula of the test microorganisms for both initial and subsequent challenges should be of sufficient concentration to provide at least 10[4] survivors on the parallel control surface.
      
      (iii) Subsequent challenges should be of sufficient frequency to accurately represent normal conditions of use. 
      
      (iv) Quantitative bacteriological sampling should be conducted at frequent and regular intervals for the length of time the residual activity can be expected to exist under the expected use conditions.
      
      (v) The same types of surfaces without the treatment should be em - ployed in the test and inoculated in a manner and over an exposure period identical to the use pattern for which the product is intended.
      
      (vi) The environmental conditions (e.g., relative humidity and temperature) should be the same as those likely to be encountered under normal conditions of product use. Tests should also include those environmental conditions that would act to reduce the effective concentration of the product on the inani - mate surface (e.g., rinsing, abrasion, organic load, repeated challenges by microorganisms, etc.).
      
      (vii) The length of time the residual activity can be expected to exist under the expected use conditions should be documented.
      
      (2) Evaluation of success of residual self-sanitizing action. For residual self-sanitizing claims, it should be demonstrated that a product is capable of reducing the number of test microorganisms on the test surface by 99.9% over that of the parallel control surfaces.
      
      (l) Residual self-sanitizing activity of dried chemical residues on hard-inanimate surfaces - dry surfaces. This section addresses efficacy testing for products which bear label claims to provide residual self-sanitizing activity (e.g., significant reduction in numbers of infectious microorganisms which may be present or subsequently deposited) on treated dry surfaces.
      
      (1) Test Procedure. The Agency recommends the use of the Protocol for Residual Self-Sanitizing Activity of Dried Chemical Residues on Hard Non-Porous Surfaces. This guidance may be found at:  http://www.epa.gov/oppad001/regpolicy.htm.
      
      (2) Evaluation of residual self-sanitizing success. The product should demonstrate that it is capable of reducing the number of test microorganisms on the test surface by >=99.9% over that of the parallel control surfaces in >=5 minutes for a 24 hour period.
      
      (m)  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. Where recommended methods are modified to support specific claims and/or use patterns for a product, the protocol, identifying and describing each modification, should be provided in specific detail with the test 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 to submit the proposed testing protocols for in-use or simulated-use studies to the Agency for review and evaluation prior to initiation of the test.
      
      (n) References: The following references may be consulted for additional background information:
      
      (1) Annual Book of Standards, Test Method for Efficacy of Sanitizers Recommended for Inanimate Non-Food Contact Surfaces, Designation E1153. American Society for Testing and Materials, 100 Barr Harbor Drive, West Conshohocken, PA 19428.
      
      (2) Official Methods of Analysis of the AOAC International, Chapter 6, Disinfectants, Official Method 955.16 Chlorine (Available) in Disinfectants, Germicidal Equivalent Concentra - tion. Current edition. AOAC International, Suite 500, 481 North Frederick Avenue, Gaithersburg, MD 20877-2417.
      
      (3) Official Methods of Analysis of the AOAC International, Official Method 960.09 Germicidal and Detergent Sanitizing Action of Disinfectants. Current edition. AOAC International, Suite 500, 481 North Frederick Avenue, Gaithersburg, MD 20877-2417.