&EPA
         United States
         Environmental Protection
         Agency
          Office of Chemical Safety and
          Pollution Prevention
          (751 OP)
EPA712-C-07-091
 January (insert
  date), 2012
Product Performance
Test Guidelines
OCSPP 810.2300:
Sanitizers for Use on
Hard Surfaces—Efficacy
Data Recommendations

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                                  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.

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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.

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       (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
Non-food
Contact
Sanitizer
Food Contact
Surface
Sanitizer
Halide Products
Food Contact
Surface
Sanitizer
Non-Halide
Products
Food Contact
Surface
Sanitizer
Towelette
Sanitizers for
Urinal and Toilet
Bowl Water and
In-tank
Sanitizers
Test Methods
Water soluble
powders/liquids
Spray products
Towelettes
Water soluble
powders/liquids
Water soluble
powders/liquids
Towelettes
Water soluble
powders/liquids/tablets
ASTME-1153
Reserved
AOAC
International
Chlorine
(Available) in
Disinfectants
Germicidal
Equivalent
Concentration
AOAC
International
Germicidal and
Detergent
Sanitizing Action
of Disinfectants
http://www.epa. g
ov/oppad001/tow
elette
Simulated-use
study
Test Organisms
Staphylococcus aureus (ATCC
6538) and Klebsiella
pneumoniae (ATCC 4352)
Enterobacter aerogenes
(ATCC 13048) may be
substituted for K. pneumoniae.
Salmonella enterica (formerly
typhi) (ATCC 6539) or S.
aureus (ATCC 6538)
Escherichia coli (ATCC 11229)
and S. aureus (ATCC 6538)
Enterococcus faecium ATCC
6569 or Salmonella enterica
(formerly choleraesuis) ATCC
10708
No. of
Batches/Carriers
Three batches, one at least 60
days old.
Three batches, one at least 60
days old.
Three batches, one at least 60
days old.
Three batches, one of which is at
least 60 days old.
Evaluation
of Success
99.9% reduction
within 5 minutes.
Test results should
demonstrate product
concentrations
equivalent in activity
to 50, 100, and 200
ppm of available
chlorine.
99.999% reduction
in the number of
each test
microorganism
within 30 seconds.
99.9% reduction
over parallel control
counts.

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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 (El 153) 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-logio 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

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Inanimate Non-Food Contact Surfaces (El 153) 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.  entericd)  (formerly Salmonella typhf)
(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 quaternary
ammonium  compounds,   chlorinated  trisodium  phosphate  and   anionic  detergent-acid
formulations.

       (i) Test procedure. The Agency  recommends the AOAC International Germicidal and
Detergent Sanitizing Action of Disinfectants test (Ref.  3). Three samples, representing  three

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different batches, one of which should be at least 60 days old, should be evaluated for efficacy
against both Escherichia coll (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  Detergent  Sanitizing  Action  of
Disinfectants test (Ref.  3). One sample should be  evaluated for  efficacy  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 (PCS) 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. PCS 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.

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       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)(l)(i) through (i)(l)(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 documented.

       (iii)  Inocula containing representative Gram-positive (e.g., Enterococcus faecium ATCC
6569) or Gram-negative (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 104 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 untreated 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.

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       (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)(l)(i) through (j)(l)(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)(l)(i)(A) and  (j)(l)(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)(l)(i)(A) in this guideline to demonstrate the pH and concentration of the active
 ingredients.

       (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 demonstrating 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
 104CFU/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)(l)(i) through (k)(l)(vii) of this guideline.

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       (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 104 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 employed 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 inanimate 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.

       (1) 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.

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       (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);

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       (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  El 153.  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,
                                           10

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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.
                                           11

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