STANDARDS | ECO SEAL CANADA

Cleaning and De-greasing Compounds: Biologically-based

Introduction

The Eco-Seal Program is pleased to publish the following certification criteria document on biologically-based cleaning and degreasing compounds.

The Eco-Seal Program is designed to support a continuing effort to improve and/or maintain environmental quality by reducing energy and materials consumption and by minimizing the impacts of pollution generated by the production, use and disposal of goods and services.

Industrial and commercial cleaners are used primarily for general facility cleaning and machinery/ parts cleaning. The selection of a cleaner is influenced primarily by the nature of the surface to be cleaned, the nature of the soil, and the degree of cleanliness required. The active ingredients in conventional cleaner formulations are surfactants, builders, alkalis and organic solvents. Conversely, biologically-based cleaning and degreasing compounds contain enzymes and/or microbial cultures that promote microbial digestion of hydrocarbons, organic contaminants and other undesirable substances. Biologically-based products may also contain

surfactants and other compounds that increase their efficacy.

Surfactants lower the surface tension of water and allow the cleaning solution to penetrate and suspend soils. If surfactants are not easily biodegraded they may persist and harm ecosystems. The products of degradation should also not pose an elevated risk to the environment.

Builders serve to control water hardness and improve surfactant performance. Builders may have adverse impacts on aquatic systems and water quality if present at excessive concentrations. The use of aqueous or semi-aqueous cleaners rather than those made primarily of organic solvents results in a reduction in volatile organic compounds and ozone depleting substances emitted.

In order to be eligible to carry the Eco-Seal, the biologically-based cleaning and degreasing compound must:

● be biologically based,

● not be toxic,

● not be harmful to humans, riparian species and aquatic species,

● not accumulate in the environment,

● be accompanied by instructions for use and disposal, and

● be efficacious, as determined by testing comparable to chemically-based products.

Based on a review of currently available life cycle information, the category requirements will produce an environmental benefit through the reduction of harmful effluents and wastewater loading, and resource conservation.

Certification Criteria Document Cleaning and De-greasing Compounds:

Biologically-based

Life cycle review is an ongoing process. As information and technology change, the category requirements will be reviewed and possibly amended.

Notice

Any reference to a standard means to the latest edition of that standard.

The Eco-Seal Program reserves the right to accept equivalent test data for the test methods specified in this document.

Interpretation

1. In this criteria document:

“APEO” means alkylphenol ethoxylate;

“aqueous cleaner” means a hard-surface cleaning formulation in which water is the main solvent, including alkaline cleaners and surfactant concentrates. For the purposes of this guideline, acid cleaners are not included;

“ATCC” means American Type Culture Collection, and is a biotechnology organization that has a diverse repository of microorganisms, cell lines and molecular biology materials;

“biodegradable” means readily biodegradable, as determined using one of the methods described in OECD Guidelines for the Testing of Chemicals, provided that all measurements and calculations are based on the carbon content of the mixture and its degradation, i.e., dissolved organic carbon (DOC) removal (301A or 301E), CO2 evolution (301-B) or oxygen consumption in the presence of an inhibitor of nitrogen metabolism (301C, 301D or 301F);

“Biosafety Level” means a classification of biological safety that may be documented on a material safety data sheet (MSDS) for a microbial culture. ATCC has the following three classifications for microbial cultures:

! biosafety level 1: the culture is not known to cause disease in healthy human adults or animals,

! biosafety level 2: the culture may cause disease in humans or animals, and

! biosafety level 3: the culture is known to cause serious or potentially lethal disease in humans

or animals; and

! biosafety level 4: dangerous/exotic agents which pose high risk of life-threatening disease,

aerosol-transmitted lab infections; or related agents with unknown risk of transmission

“builder” means any substance intended to maintain alkalinity, and/or bind dissolved metal ions (soften the water), and/or keep the soil in suspension. It includes substances such as sodium citrate, sodium silicate, phosphates, NTA, and EDTA;

“CFC” means chlorofluorocarbons;

“CGSB” means Canadian General Standards Board;

“colony forming unit” means a measure of bacteria concentration assuming that each bacterium is capable of forming a colony;

“corrosive” means a product which causes irreversible tissue damage to the skin;

“EC50" means median effective concentration. It is the concentration of material that is estimated to cause some defined toxic effect to 50% of the test organisms;

“EDTA based builders” means ethylene diaminetetraacetic acid or ethylene dinitrilotetraacetic acid or any of its salts used to maintain high solution pH, bind metal ions and emulsify soil. It does not mean ethylene diaminetetraacetic acid used as a microbiological stabilizer and biocide and present in ppm concentrations at final use dilution;

“flash point” means the minimum temperature of a liquid at which the vapours given off are sufficient to form a flammable mixture with air which will ignite when exposed to an open flame in accordance with the American Society for Testing and Materials (ASTM) test method D93-80 (Pensky-Martens Closed Tester) or ASTM test method D3278-82 (Seta);

“general facility maintenance” means the cleaning of surfaces in industrial, commercial and institutional facilities, including walls, floors, and equipment surfaces. It is not typically associated with production and manufacturing processes;

“biologically-based cleaning and degreasing compounds” means aqueous cleaners and semi-aqueous cleaners for general facility maintenance or for machinery maintenance. For the purposes of this document, non-aqueous cleaners are not included.

“LC50" means median lethal concentration. It is the concentration of material that is estimated to be lethal to 50% of the test organisms;

“machinery/equipment maintenance” means the removal of organic soil and build-ups on machinery or equipment with moving parts that is directly involved in manufacturing and production processes;

“NIOSH” means the United States National Institute for Occupational Safety and Health;

“non-aqueous cleaner” means a water-free cleaning formulation consisting exclusively or predominantly of one or more organic solvents, including chlorinated hydrocarbons, non-chlorinated aliphatic and aromatic hydrocarbons, ketones, ethers and alcohols; some contain small quantities of other detergency enhancers, particularly surfactants;

“NTA” means nitrilotriacetic acid or any of its salts;

“OECD” means the Organization for Economic Co-operation and Development;

“ozone depletion potential” or “ODP” means the ratio of calculated ozone column change for each mass unit of a gas emitted into the atmosphere relative to the calculated depletion for a mass unit of the reference gas CFC-11;

“semi-aqueous cleaner” means a neutral or alkaline hard-surface cleaning formulation which combines the cleaning action of surfactants and organic solvents, including mineral spirits, glycols, glycol ethers, alcohols and terpenes, either emulsified in a water base, or in a concentrated form that can be rinsed in water;

“surfactant” (or surface-active agent) means any compound that reduces interfacial tension between two liquids or between a liquid and a solid. The three categories of surfactants are detergents, wetting agents and emulsifiers;

“toxic” means an LC50 or EC50 result for fish, daphnia, or algae of an acceptable level as determined by governmental approval of the jurisdiction in which the product is sold and/or used;

“U.S. EPA” means United States Environmental Protection Agency; and

“volatile organic compound” or “VOC” means any organic compound which participates in

atmospheric photochemical reactions. This definition excludes those organic compounds which the Eco-Seal Program designates as having negligible photochemical reactivity, a list of which appears in Appendix A.

Category Definition

2. This category includes all biologically-based cleaning and degreasing compounds as further defined in the subcategories in this section. The subcategories are:

(a) biologically based general facility maintenance cleaners (110 A); and

(b) biologically based parts cleaners (110 B).

General Requirements

3. To be authorized to carry the Eco-Seal, the biologically-based cleaning and degreasing compound must:

(a) meet or exceed all applicable governmental and industrial safety and performance standards;

And

(b) be manufactured and transported in such a manner that all steps of the process, including the disposal of waste products arising therefrom, will meet the requirements of all applicable

governmental acts, by laws and regulations.

Product Specific Requirements

4. To be authorized to carry the Eco-Seal the biologically-based cleaning and degreasing compound must:

(a) be efficacious, as determined by an appropriate test method, including:

- at least 75% cleaning efficiency as measured by test methods A5 “particulate and oily soil/vinyl tiles” or A6 ”oil, carbon, black and clay/white enamel painted stainless-steel panels” in ASTM D4488-95(2001)e1 01-Jan-1995 “Standard Guide for Testing Cleaning Performance of Products Intended for Use on Resilient Flooring and Washable Walls”, OR

- compliance with CAN/CGSB 2-GP-11, Method 20.3 “Methods of Testing and

Analysis of Soaps and Detergents: Cleaning Efficiency";

(b) be in compliance with federal legislation regarding toxicity and biodegradation, including, for Canada, the New Substances Notification Regulations as per the Canadian Environmental Protection Act, 1999;

- in the US, the Federal Hazardous Substances Act (16 CFR Part 1500), and/or

- in Canada, the Controlled Product Regulations of the Hazardous Products Act;

(d) be accompanied by detailed instructions that identify the following to maximize product performance:

(i) the product’s proper disposal and use, and

(ii) the biological component (or the product) may not be effective in the presence of chemical sanitizers such as formaldehyde or chlorine bleach;

(e) be clearly identified as a product not intended or to be sold for domestic use;

(f) not have a pH lower than 2.0 or greater than 11.5 as measured directly in a liquid formulation or as measured in a 10% weight/weight solution of a powdered formulation;

(g) if pH is greater than 11.5, not be corrosive to skin as determined by InVitro International's CORROSITEXR test;

(h) not have a flash point lower than 61°C or 142°F;

(i) have a maximum temperature usage which does not exceed 17°C or 63°F below flash point;

(j) have a zero ODP;

(k) not be formulated with:

(i) APEO,

(ii) aromatic solvents,

(iii) chlorinated organic solvents,

(iv) butoxy-ethanol,

(v) EDTA based builders,

(vi) NTA, or

(vii) phosphorus-based builders;

(l) not be formulated with an ingredient that is listed as a Group 1 (known) , Group 2A (possible), or Group 2B (probable) carcinogen in the International Agency for Research on Cancer (IARC) Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans;

(m) not contain volatile organic compounds according to the table below:

as measured by one of the following:

U.S. EPA Method 24-24A, 40 C.F.R., Part 60, Appendix A (1991),

U.S. EPA Method 18, 48 Federal Register 48, no. 202, October 18, 1983,

U.S. EPA Method 8240 GC/MS Method for Volatile Organics, September 1986,

NIOSH

Manual of Analytical Methods, Method 1400, Volume 1, February 1984, or oras demonstrated through calculation from records of the amounts of constituents used to make the product.

For products for which the label specifies dilution prior to use, VOCs should be measured after the minimum recommended dilution has taken place. The minimum recommended dilution shall not include recommendations for the incidental use of a concentrated product to deal with limited special applications, such as hard to remove soils and stains;

(n) use only those surfactants that are biodegradable and according to the table below:

(o) have either a plate count that is greater than or equal to 1 x 107 colony forming units per millilitre or have standardized test results demonstrating enzyme activity; and

(p) if derived from or using bacterial cultures, use only those bacterial cultures that are derived from a Biosafety Level 1 ATCC microbial culture (or equivalent).

Verification

5. To verify a claim that a product meets the criteria listed in the document, the Eco-Seal Program will require access, as is its normal practice, to relevant quality control and production records and the right of access to production facilities on an announced basis.

6. Compliance with section 3(b) shall be attested to by a signed statement of the Chief Executive Officer or the equivalent officer of the manufacturer. The Eco-Seal Program shall be advised in writing immediately by the licensee of any non-compliance which may occur during the term of the license. On the occurrence of any non-compliance, the license may be suspended or terminated as stipulated in the license agreement.

Conditions for Eco-Seal Use

7. The Eco-Seal may appear on wholesale or retail packaging, or on the product itself, provided that the product meets the requirements in this document.

8. It is recommended that a criteria statement appear with the Eco-Seal whenever the Eco-Seal is used in association with the biologically-based cleaning and degreasing compound. The intent of this statement is to provide clarification as to why the product was certified and to indicate constraints to which the certification is limited. This is to ensure no ambiguity over, or misrepresentation of, the reason(s) for certification.

The recommended criteria statement wording for this product type is “biologically-based cleaner/ degreaser”. The licensee may propose other wording for the criteria statement, but any such proposed wording must be approved by the Eco-Seal Program.

9. All licensees and authorized users must comply with the Program's Guide to Proper Use of the Eco-Seal regarding the format and usage of the Eco-Seal.

10. Any accompanying advertising must conform with the relevant requirements stipulated in this document, the license agreement and the Program's Guide to Proper Use of the Eco-Seal.

For additional copies of this criteria document or for more information about the Eco-Seal Program, please contact: info@eco-seal.org

Appendix A:

Volatile Organic Compounds with Negligible Photochemical Reactivity

The list of volatile organic compounds (VOCs) designated by the Eco-Seal Program as having negligible photochemical reactivity has been taken from the following two documents:

1. State of California Air Resources Board, Regulation for Reducing Volatile Organic Compound Emissions from Consumer Products, Appendix.

2. U.S. EPA VOC Definition, Federal Register, Volume 57, No. 22, 3 February 1992, Rules and Regulations, pg. 3945, sec.51.100.

This Eco-Seal designated list includes the following compounds:

(a) acetone

(b) ammonium carbonate

(d) carbonic acid

(e) ethane

(f) metallic carbides or carbonates

(g) methane

(h) methylene chloride (dichloromethane)

(i) cyclic, branched, or linear completely methylated siloxanes

(j) parachlorobenzotrifluoride (PCBTF)

(k) perchloroethylene (tetrachloroethylene)

(l) 1,1,1-trichloroethane

(m) trichlorofluoromethane (CFC-11)

(n) dichlorodifluoromethane (CFC-12)

(o) trichlorotrifluoroethane (CFC-113)

(p) dichlorotetrafluoroethane (CFC-114)

(q) chloropentafluoroethane (CFC-115)

(r) chlorodifluoromethane (HCFC-22)

(s) dichlorotrifluoroethane (HCFC-123)

(t) dichlorofluoroethane (HCFC-141b)

(u) chlorodifluoroethane (HCFC-142b)

(v) 2-chloro-1,1,1,2-tertrafluoroethane

(HCFC-124)

(w) trifluoromethane (HFC-23)

(x) 1,1,1,2,3,4.4,5,5,5-decafluoropentane (HFC-43-10mee)

(y) pentafluoroethane (HFC-125)

(z) 1,1,2,2-tetrafluoroethane (HFC-134)

(aa) tetrafluoroethane (HFC-134a)

(bb) 1,1,1-trifluoroethane (HFC-143a)

(cc) 1,1-difluoroethane HFC-152a)

(dd) 3,3-dichloro-1,1,1,2,2-pentafluoropropane (HCFC-225ca)

(ee) 1,3-dichloro-1,1,2,2,3-pentafluoropropane (HCFC-225cb)

(ff) perfluorocarbons (classes of):

(A) cyclic, branched, or linear, completely fluorinated alkanes

(B) cyclic, branched, or linear, completely fluorinated ethers with no unsaturations

(D) sulfur-containing perfluorocarbons with no unsaturations with the sulfur bonds only to carbon and fluorine

Eco-Seal General Screen for Safer Ingredients

1 Introduction

The Eco-Seal General Screen for Safer Ingredients is a comprehensive, science-based tool designed to ensure that the safest possible ingredients are used in Eco-Seal-recognized products. The contents of this screen, including definitions and toxicological preferences were developed to facilitate use of safer chemistry under the Eco-Seal Program.

In Eco-Seal’s Product Recognition Program, the General Screen serves as the primary tool to advance Green Chemistry in product formulation and to implement Informed Substitution, the reasoned transition from a chemical of particular concern to safer chemicals or non-chemical alternatives. Eco-Seal will use the general screen to review all product ingredients (and their components) for which a more customized screen has not been developed.

The Eco-Seal Program provides a unique approach to product review and recognition. Eco-Seal evaluates each and every ingredient in a formulation within its functional class context (e.g., surfactants, solvents) and based on its key, distinguishing, human health and environmental characteristics. In this way, potential product ingredients can be viewed as part of a continuum of improved or safer ingredient choices.

The General Screen makes it possible to draw a line demarcating the greener or “low-concern” end of the continuum. To define low-concern, Eco-Seal uses toxicological thresholds established by several highly respected health and environmental protection authorities ; namely, the United Nation’s Globally Harmonized System (GHS) for the Classification and Labelling of Hazard Substances.

2 Preferences and Terms

The following preferences and terms apply to all attributes and data requirements.

2.1 A component is a chemical as identified by its Chemical Abstract Service (CAS) number.

2.2 An ingredient may be one component or a blend of multiple components.

2.3 Test data using dermal and inhalation exposure routes are preferred over oral exposure data because the former are more likely routes of exposure for cleaning products.

2.4 Data for all available routes of exposure will be evaluated. Failure to pass an endpoint by any route of exposure results in failure to pass the screen.

2.5 The GHS criteria and data evaluation approach will inform professional judgment in the review of both no observed adverse effect levels/concentrations (NOAEL/NOAEC) and lowest observed adverse effect levels/concentrations (LOAEL/LOAEC). NOAEL/NOAEC and LOAEL/LOAEC values are preferred to no observed effect levels/concentrations (NOEL/NOECs) and lowest observed effect levels/concentrations (LOEL/LOECs).

2.6 Use of existing data should follow the EPA HPV OECD HPV Programme data adequacy guidelines.

3 Attributes of Concern for all Components

Each attribute listed below applies to all components.

3.1 ACUTE MAMMALIAN TOXICITY

3.1.1 Criteria and Data Evaluation

To be acceptable under the screen, components must have a median lethal dose or concentration greater than those values listed in Table 1. Data must be available for at least one route of exposure. For inhalation, exposure should be at least four hours; the thresholds for inhalation are the same for exposures greater than four hours. Exposures of less than four hours will be evaluated on a case-bycase basis. Data for all available routes of exposure will be evaluated. Failure to pass this endpoint by any route of exposure results in failure to pass the screen.

Table 1 – Acute Mammalian Toxicity

3.1.2 Test Methods

– OPPTS Harmonized Guideline: 870.1100 Acute oral toxicity [1];

– OPPTS Harmonized Guideline: 870.1200 Acute dermal toxicity [2];

– OPPTS Harmonized Guideline: 870.1300 Acute inhalation toxicity [3];

– OECD Test Guideline 420: Acute Oral Toxicity – Fixed Dose Method [4];

– OECD Test Guideline 423: Acute Oral Toxicity – Acute Toxic Class Method [5];

– OECD Test Guideline 425: Acute Oral Toxicity – Up-and-Down Procedure [6];

– OECD Test Guideline 402: Acute Dermal Toxicity [7]; and

– OECD Test Guideline 403: Acute Inhalation Toxicity [8].

3.2 CARCINOGENICITY

3.2.1 Criteria and Data Evaluation

Components will be screened for carcinogenicity based upon established lists (see Table 2) and GHS criteria. Components not on the established lists in Table 2, but that are considered known or presumed human carcinogens (Category 1), or suspected human carcinogens (Category 2) under GHS [9], will not pass the screen. Available data on the component or a chemically (e.g., based on chemical structure) or biologically (e.g., based on metabolic breakdown, or likely mechanistic/mode of action considerations) similar analog along with OncoLogic™ [10] will be used to assess a chemical under GHS.

Table 2 – Carcinogenicity

3.2.2 Preferred Test Methods

– OECD Test Guideline 451: Carcinogenicity Studies [12]

– OECD Test Guideline 453: Combined Chronic Toxicity/Carcinogenicity Studies [13]

– OPPTS Harmonized Guideline 870.4200: Carcinogenicity [14]

– OPPTS Harmonized Guideline 870.4300: Combined Chronic Toxicity/Carcinogenicity [15]

– NTP 2 year Study Protocol “Specifications for the conduct of studies to evaluate the toxic and carcinogenic potential of chemical, biological and physical agents in laboratory animals for the National Toxicology Program” [16].

3.2.3 Data Interpretation

– Section 2, Hazard Assessment in Guidelines for Carcinogen Risk Assessment [17]

– GHS Ch 3.6 Carcinogenicity [9]

3.3 ENVIRONMENTAL TOXICITY AND FATE

3.3.1 Criteria and Data Evaluation

If a component is an acute aquatic toxicant, then it must biodegrade rapidly and not be bioaccumulative (see Table 3, lines 1-3). If a component has low aquatic toxicity (Table 3, line 4), then its rate of biodegradation may be >28 days.

Table 3 – Environmental Toxicity and Fate

3.3.2 Test Methods, Acute Aquatic Toxicity

A baseline data set is required that should include freshwater test data for a species each of algae, aquatic invertebrate and fish. Additional aquatic toxicity data in other species or in marine species will also be reviewed if available.

3.3.2.1 Preferred test methods for fish

– OECD Test Guideline 203: Fish, Acute Toxicity Test [19]

– OPPTS Harmonized Guideline 850.1075: Fish acute toxicity test, freshwater and marine [20]

3.3.2.2 Preferred test methods for aquatic invertebrates

– OECD Test Guideline 202, Part 1, Daphnia sp., Acute Immobilisation Test [21]

– OPPTS Harmonized Guideline 850.1010: Aquatic invertebrate acute toxicity test, freshwater daphnids [22]

– OPPTS Harmonized Guideline 850.1035: Mysid acute toxicity test [23]*

*NOTE – A 96 hour Mysid shrimp acute toxicity test can be used in place of a daphnid acute toxicity test when the latter is not available.

3.3.2.3 Preferred test methods for aquatic plants

– OECD Test Guideline 201, Alga, Growth Inhibition Test [24]

– OPPTS Harmonized Guideline 850.5400: Algal toxicity, Tiers I and II [25]

3.3.2.4 Alternative test methods for acute aquatic toxicity

The following test methods may be considered, when relevant:

– OPPTS Harmonized Guideline 850.1085: Fish acute toxicity mitigated by humic acid [26]

– OPPTS Harmonized Guideline 850.1025: Oyster acute toxicity test (shell deposition) [27]

– OPPTS Harmonized Guideline 850.1045: Penaeid acute toxicity test [28]

– OPPTS Harmonized Guideline 850.1055: Bivalve acute toxicity test (embryo larval) [29]

– OPPTS Harmonized Guideline 850.4400: Aquatic plant toxicity test using Lemna spp.

Tiers I and II [30]

– Modeled data from sources such as ECOSAR [31] are acceptable when data are unavailable

3.3.3 Test Methods, Persistence (measured as biodegradation)

Data from experimental methods is generally preferred over estimations of persistence. For the purposes of screening safer chemicals, ready biodegradation tests are preferred. It is noted that simulation tests are likely to better describe the biodegradability of a particular chemical in specific environmental conditions.

3.3.3.1 Preferred test methods

– OECD Test Guideline 301: Ready Biodegradability (sections A-F [32])

– OPPTS Harmonized Guideline 835.3110: Ready biodegradability [33]

3.3.3.2 Alternative test methods

– Modeled data from sources such as EPISuite [34] and the PBT Profiler [35] are acceptable when data are unavailable.

3.3.4 Test Methods, Bioaccumulation

3.3.4.1 Preferred test methods

A field-measured BAF (located in the literature) is the most preferred data for indicating bioaccumulation. When not possible, the following test methods may be used:

– OECD Test Guideline 305: Bioconcentration: Flow-through Fish Test [36]

– OPPTS Harmonized Guideline 850.1710: Oyster BCF [37]

– OPPTS Harmonized Guideline 850.1730: Fish BCF [38]

3.3.4.2 Alternative test methods

– Modeled data from sources such as EPISuite [34] and the PBT Profiler [35] are acceptable when data are unavailable. An estimated BAF is preferred to an estimated BCF for compounds where log Kow > 5.

3.4 GENETIC TOXICITY

3.4.1 Criteria and Data Evaluation

Components will be screened for mutagenicity based upon test data (see Table 4 for details). Data on genetic toxicity must be available for at least one potential mutagenic effect. Data for all available effects will be evaluated. Effects to be considered are: heritable germ cell mutagenicity, germ cell genetic toxicity, and somatic cell mutagenicity or genetic toxicity. Data from in vivo, in vitro, and epidemiological

studies will be considered.

Table 4 – Genetic Toxicity

3.4.2 Preferred Test Methods

– OECD Test Guideline 474: Mammalian Erythrocyte Micronucleus Test [40];

– OECD Test Guideline 475: Mammalian Bone Marrow Chromosome Aberration Test [41];

– OECD Test Guideline 478: Genetic Toxicology: Rodent Dominant Lethal Test [42];

– OECD Test Guideline 479: Genetic Toxicology: In Vitro Sister Chromatid Exchange Assay in Mammalian Cells [43];

– OECD Test Guideline 483: Mammalian Spermatogonial Chromosome Aberration Test [44];

– OECD Test Guideline 484: Genetic Toxicology: Mouse Spot Test [45];

– OECD Test Guideline 485: Genetic Toxicology: Mouse Heritable Translocation Assay[46];

– OPPTS Harmonized Guideline 870.5195: Mouse Biochemical Specific Locus Test [47];

– OPPTS Harmonized Guideline 870.5200: Mouse Visible Specific Locus Test [48];

– OPPTS Harmonized Guideline 870.5380: Mammalian Spermatogonial Chromosome Aberration Test [49];

– OPPTS Harmonized Guideline 870.5385: Mammalian Bone Marrow Chromosome Aberration Test [50];

– OPPTS Harmonized Guideline 870.5395: Mammalian Erythrocyte Micronucleus Test [51];

– OPPTS Harmonized Guideline 870.5450: Rodent Dominant Lethal Assay [52]; and

– OPPTS Harmonized Guideline 870.5460: Rodent Heritable Translocation Assays [53].

3.4.3 Acceptable Test Methods

Per GHS [39], results from multiple acceptable test methods must be used in conjunction for evaluation of genetic toxicity. In some cases, the results from acceptable test methods must be used in conjunction with preferred test methods. The following test methods are acceptable:

– OECD Test Guideline 471: Bacterial Reverse Mutation Test [54];

– OECD Test Guideline 473: In Vitro Mammalian Chromosome Aberration Test [55];

– OECD Test Guideline 476: In Vitro Mammalian Cell Gene Mutation Test [56];

– OECD Test Guideline 486: Unscheduled DNA Synthesis (UDS) Test with Mammalian Liver Cells In Vivo [57];

– OPPTS Harmonized Guideline 870.5100: Bacterial Reverse Mutation Test [58];

– OPPTS Harmonized Guideline 870.5300: In Vitro Mammalian Cell Gene Mutation Test [59];

– OPPTS Harmonized Guideline 870.5375: In Vitro Mammalian Chromosome Aberration Test [60];

– OPPTS Harmonized Guideline 870.5500: Bacterial DNA Damage or Repair Tests [61];

– OPPTS Harmonized Guideline 870.5550: Unscheduled DNA Synthesis in Mammalian Cells in Culture [62];

– OPPTS Harmonized Guideline 870.5900: In Vitro Sister Chromatid Exchange Assay [63];

and

– OPPTS Harmonized Guideline 870.5915: In Vivo Sister Chromatid Exchange Assay [64].

3.4.4 Data Interpretation

GHS Ch. 3.5 Germ Cell Mutagenicity [39]

3.5 NEUROTOXICITY

3.5.1 Criteria and Data Evaluation

Components that are considered neurotoxicants under GHS [9] criteria (see guidance values in Table 5) will not pass the screen. Available data on the component or a chemically (e.g., based on chemical structure) or biologically (e.g., based on metabolic breakdown, or likely mechanistic/mode of action considerations) similar analog will be used to assess a solvent under GHS. Insufficiently characterized chemicals may be considered for the Eco-Seal Screen.

Table 5 – Neurotoxicity

3.5.2 Preferred Test Methods

– OECD Test Guideline 424: Neurotoxicity Study in Rodents [65] and

– OPPTS Harmonized Guideline 870.6200: Neurotoxicity screening battery [66].

3.5.3 Optional Test Methods

Additional evidence from OECD Test Guideline 426: Developmental Neurotoxicity Study [67] and OPPTS Harmonized Guideline: 870.6300 Developmental neurotoxicity study [68] can be used to screen components for neurotoxicity.

3.5.4 Data Interpretation

– Section 3, Hazard Characterization in Guidelines for Neurotoxicity Risk Assessment [69]

– GHS Ch. 3.9 Specific Target Organ Toxicity Repeated Exposure [70]

3.6 REPEATED DOSE TOXICITY

3.6.1 Criteria and Data Evaluation

Components that are considered systemic toxicants under GHS [70] (see guidance values in Table 6) will not pass the screen. Data for all available routes of exposure will be evaluated, and any study must be 28 days or greater to satisfy this endpoint. Should testing be pursued to meet the screen data requirement, a functional observational battery (FOB) should be added to the test method to provide neurotoxicity information.

Table 6 – Repeated-Dose Toxicity

3.6.2 Preferred Test Methods

– OECD Test Guideline 408: Repeated Dose 90-Day Oral Toxicity Study in Rodents [71]

– OECD Test Guideline 409: Repeated Dose 90-Day Oral Toxicity Study in Non-Rodents [72]

– OECD Test Guideline 411: Subchronic Dermal Toxicity: 90-day Study [73]

– OECD Test Guideline 413: Subchronic Inhalation Toxicity: 90-day Study [74]

– OPPTS Harmonized Guideline 870.3100: 90-Day oral toxicity in rodents [75]

– OPPTS Harmonized Guideline 870.3150: 90-Day oral toxicity in nonrodents [76]

– OPPTS Harmonized Guideline 870.3250: 90-Day dermal toxicity [77]

– OPPTS Harmonized Guideline 870.3465: 90-Day inhalation toxicity [78]

3.6.3 Acceptable Test Methods

– OECD Test Guideline 412: Repeated Dose Inhalation Toxicity: 28-day Study [79]

– OECD Test Guideline 410: Repeated Dose Dermal Toxicity: 28-day Study [80]

– OECD Test Guidelines 407: Repeated Dose 28-day Oral Toxicity Study in Rodents [81]

– OECD Test Guideline 422, Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test [82]

– OPPTS Harmonized Guideline 870.3050: Repeated dose 28-day oral toxicity study in rodents [83]

– OPPTS Harmonized Guideline 870.3200: 28-Day dermal toxicity [84]

3.6.4 Data Interpretation

GHS Specific Target Organ Toxicity – Repeated Exposure [70]

3.7 REPRODUCTIVE AND DEVELOPMENTAL TOXICITY

3.7.1 Criteria and Data Evaluation

Components that are considered reproductive or developmental toxicants under GHS [85] (see guidance values in Table 7) will not pass the screen. Following the SIDS Dossier [86], all chemicals must be reviewed for both fertility and developmental effects. Data on reproductive and developmental toxicity must be available via at least one route of exposure, oral, dermal, or inhalation. Data for all available routes of exposure will be evaluated. Failure to pass this endpoint by any route of exposure or toxic effect (fertility or development) results in failure to pass the screen.

Table 7 – Reproductive and Developmental Toxicity

3.7.2 Test Methods

3.7.2.1 Fertility test methods, preferred

– OECD Test Guideline 415: One-Generation Reproduction Toxicity Study [87] and

– OECD Test Guideline 416: Two-Generation Reproduction Toxicity Study [88].

3.7.2.2 Fertility test methods, acceptable

The following test methods may be used to identify reproductive toxicity per GHS [85]:

– OPPTS Harmonized Guideline 870.3800: Reproduction and fertility effects [89];

– OECD Test Guideline 421: Reproduction/Developmental Toxicity Screening Test [90];

– OECD Test Guideline 422: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test [82];

– OPPTS Harmonized Guideline 870.3550: Reproduction/developmental toxicity screening test [91]; and

– OPPTS Harmonized Guideline 870.3650:Combined repeated dose toxicity study with the reproduction/developmental toxicity screening test [92].

3.7.2.3 Developmental toxicity test methods, preferred

– OECD Test Guideline 414: Prenatal Developmental Toxicity Study [93].

3.7.2.4 Developmental toxicity test methods, acceptable

The following test methods may be used to identify developmental toxicity per GHS [85]:

– OPPTS Harmonized Guideline 870.3800: Reproduction and fertility effects [89];

– OECD Test Guideline 421: Reproduction/Developmental Toxicity Screening Test [90];

– OECD Test Guideline 422: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test [82];

– OPPTS Harmonized Guideline 870.3550: Reproduction/developmental toxicity screening test [91]; and

– OPPTS Harmonized Guideline 870.3650:Combined repeated dose toxicity study with the reproduction/developmental toxicity screening test [92].

3.7.3 Data Interpretation

– Section 3, Hazard Characterization, Guidelines for Reproductive Toxicity Risk Assessment [94]

– Section 3, Hazard Characterization, Guidelines for Developmental Toxicity Risk Assessment [95]

– GHS Ch 3.7 Reproductive Toxicity [85]

3.8 RESPIRATORY SENSITIZATION

3.8.1 Criteria and Data Evaluation

Although recognized animal models for the testing of respiratory hypersensitivity are not available at present, the following will be applied when models are available. Components that are considered respiratory sensitizers under GHS [96] will not pass the screen. Data on respiratory sensitization will normally be based on human experience. Hypersensitivity may be demonstrated by clinical history with supporting tests, positive bronchial challenge tests, or appropriate animal studies. There is currently no preferred test method, so evaluation will be based on all valid, available data.

3.8.2 Data Interpretation

GHS Ch. 3.4 Skin or Respiratory Sensitization [96]

3.9 SKIN SENSITIZATION

3.9.1 Criteria and Data Evaluation

Components that are considered skin sensitizers under GHS [96] will not pass the screen. Available data for the component and analogs will be used to assess a component under GHS.

3.9.2 Preferred Test Methods

– OECD Test Guideline 406: Skin Sensitization [97]

– OECD Test Guideline 429: Skin Sensitization: Local Lymph Node Assay [98]

– OPPTS Harmonized Guideline 870.2600: Skin Sensitization [99]

3.9.3 Data Interpretation

GHS Ch. 3.4 Skin or Respiratory Sensitization [96]

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98. OECD, OECD Guidelines for the Testing of Chemicals: Test No. 429: Skin Sensitization: Local Lymph Node Assay. 2002.

99. USEPA, Health Effects Test Guidelines: OPPTS 870.2600: Skin Sensitization. 1998.