http://www.epa.gov/oppt/newchem s/chem cat.htm



# TSCA NEW CHEMICALS PROGRAM (NCP) CHEMICAL CATEGORIES

Contact: Office of Pollution Prevention and Toxics

U.S. Environm ental Protection Agency 1200 Pennslyvania Avenue, N.W W a shington, D.C. 20460 Telephone: (202) 564-8996 Miriam W i ggins-Lewis (m ail code 7405M; wigginslewis.m i riam @epa.gov) J. Vincent Nabholz (m ail code 7403M; nabholz.joe@epa.gov)

Rebecca Jones (m ail code 7403M; jones.rebecca@epa.gov) Last revised: October, 2002Note on Test Guidelines Cited in this Document: This docum ent contains references to test guidelines that originally appeared in title 40, chapter I, subchapter R of the Code of Federal Regul ations (CFR). Since that tim e, the "OPPTS Harm onized Test Guidelines" have been developed by EPA' s Office of Prevention, Pesticides and Toxic Substances for use in the testing of pesticides and toxic substances, and the developm ent of test data that m u st be subm itted to the Agency for review under Federal regulations. These guidelines can be found at http://www.epa.gov/docs/OPPTS_Harm onized/. Until we fully update all the references contained within this categories docum ent, you can find any new OPPTS guideline that corresponds to an old CFR citation. For exam ple, the fish acute toxicity test at 40 CFR 797.1400 has been replaced by OPPTS 850.1075, and Ready Biodegradability at 40 CFR 796.3180, etc. has been replaced by OPPTS 835.3110. Disclaimer: EPA' s Office of Pollution Prevention and Toxics (OPPT) groups chem icals with shared chem ical and toxicological properties into categories so that PMN reviews are f acilitated. PMN subm itters and EPA reviewers benefit from the accum u lated data and past decisions represented by a category. EPA considers all PMNs, including new chem ical substances which fall within such categories, on a case-by-case basis and uses the m o st appropriate structural analogue to support any concerns for health or environm ental effects. For any new chem ical substance, if EPA determ ines, under TSCA section 5(e), that the substance "m ay present an unreasonable risk of injury to health or the environm ent,” the PMN substance can be subjected to appropriate control actions and/or testing under TSCA section 5(e). This “m ay present” finding is developed through the application of category-based or chem ical-by-chem ical assessm ent of hazard endpoints and case-specific exposure assessm ent. The categories included in this com p ilation represent chem icals for which sufficient assessm ent experience has been accum u lated so that hazard concerns and testing recomm e ndations vary little from chem ical to chem ical within the category. Thus, these categories do not necessarily represent the chem icals of greatest concern to the Agency. By the sam e token, the categories are also not intended to be a com p rehensive list of all substances that m ay be subject to further action in the New Chem icals Program

. If

a new chem ical substance is structurally sim ilar to a substance f o r which EPA has positive toxicity data and there is sufficient exposure, EPA m ay regulate that substance under section 5(e) of the Toxic Substances Control Act (TSCA) based on its potential unreasonable risk. The hazard potential of a PMN substance is dependent on the m o lecular weight of the com pound, the presence/absence of activating/deactivating substituents, other physical/chem i cal properties, and the likely route of exposure. Typically, concerns are confined to chem icals with m o lecular weights <1,000 whenever inhalation exposure to hum ans or environm ental release is expected, and to species <500 when derm al exposure to hum ans is expected. Additional inform ation on testing conditions and testing options for specific PMN chem ical substances that appear to fall within one of these categories is available by contacting the New Chem icals Branch at 202-260-3725 or the TSCA Hotline at 202-554-1404. This is to serve only

iias a guide f o r PMN subm itters. PMN subm itters are urged to have protocols and testing schem e s evaluated by the New Chem icals Program prior to comm e ncing testing on a PMN substance. However, under no circum stance does such an evaluation m ean that the new chem ical substance is not subject to further review during any TSCA section 5 review period for the PMN substance or that additional testing or regulatory action m ay not be recomm e nded. Finally, even if EPA does not m ake the potential unreasonable risk finding for a new chem ical substance, the Agency m ay still recomm e nd the use of engineering controls or personal protective equipm ent to reduce exposures in the workplace if the PMN substance is a potential irritant or sensitizer.

iiiTable of Contents

A c i d C h l o r i d e s ................................................................1 Acid Dyes and Am photeric Dyes .................................................2 A c ry l am i d e s ..................................................................5 A c ry l a t e s / M e th a c ry l a t e s ........................................................8 Aldehydes ..................................................................1 0 A l i p h a t i c A m i n e s .............................................................1 4 Alkoxysilanes ................................................................1 6 Alum inum Com pounds ........................................................1 8 Am inobenzothiazole Azo Dyes ..................................................2 3 Anhydrides, Carboxylic Acid ...................................................2 5 Anilines ....................................................................2 6 Dianilines .............................................................3 0 A n i o n i c S ur f a c t an t s ...........................................................3 4 A z i d e s .....................................................................3 6
B e n z o tr i a z o l e s ...............................................................3 9
B e n z o tr i a z o l e - h i n d e r e d p h e n o l s .................................................4 2 Boron Com pounds ............................................................4 5 C a t i o n i c D y e s ................................................................5 0 C a t i o n i c ( qu a t e rn ary amm o n i um ) s ur f a c t an t s .......................................5 1 C o b a l t ......................................................................5 2
D i a z o n i um s .................................................................5 6
D i c h l o r o b e n z i d i n e - b a s e d P i g m e n t s ...............................................6 0

ivD i th i o c ar b am a t e s .............................................................6 2 Epoxides ....................................................................6 4
E s t e r s ......................................................................6 5
E th y l e n e G l y c o l E th e r s ........................................................6 8 Hydrazines and Related Com pounds ..............................................7 0
H i n d e r e d A m i n e s .............................................................7 3
I m i d e s ......................................................................7 4
D i i s o c y an a t e s ................................................................7 7 

### $

- N ap h th y l am i n e s , S u l f o n a t e d ..................................................8 0
L an th an i d e s o r R ar e E arth M e t a l s ................................................8 2
N e utr a l O r g an i c s .............................................................8 6 Nickel Com pounds ............................................................8 8
N o n i o n i c S ur f a c t an t s ..........................................................9 0 O r g an o t i n s ..................................................................9 2
P e r o x i d e s ...................................................................9 6 Persistent, Bioaccum u lative,

an d T o x i c ( P B T ) C h e m i c a l s...................................9 8
P h e n o l p h th a l e i n s ............................................................1 0 1
P h e n o l s ....................................................................1 0 3
P h o s p h a t e s , I n o r g an i c ........................................................1 0 6
P h o s p h i n a t e E s t e r s ...........................................................1 0 9 Polyanionic Polym e rs (& Monom ers) ............................................1 1 2
P o l y c a t i o n i c P o l y m e r s ........................................................1 1 3
P o l y n i tr o ar o m a t i c s ...........................................................1 1 4

vR e s p i r ab l e , P o o r l y S o l ub l e P art i c u l a t e s ...........................................1 1 8
R o s i n .....................................................................1 2 3 Stilbene, derivatives of 4,4-bis(triazin-2-ylam ino)- .................................1 2 5
T h i o l s .....................................................................1 2 8 Substituted Triazines .........................................................1 3 1 Triarylm ethane Pigm ents/Dyes with Non-solubilizing Groups ...............................1 3 2
V i ny l E s t e r s ................................................................1 3 7 V i ny l S u l f o n e s ..............................................................1 3 9 S o l ub l e c o m p l e x e s o f Z i n c .....................................................1 4 3 Zirconium Com pounds .......................................................1 4 4

viCategory: Acid Chlorides Environm ental Toxicity Definition

. This category includes carbonyl chlorides (R-C[=O]Cl) and sulfochlorides (RS[ =O] C l) where R m ay be either aliphatic or arom atic. Toxicity is lim ited by the f act that this class of com pounds hydrolyzes and also, probabl y, if the octanol/water partition coefficient (Kow) is above a log Kow value of 8. It has been assum e d that these com pounds need to be absorbed to be toxic, therefore, com pounds with MW s > 1000 will probably be excluded in the future once this assum p tion is confirm e d with toxicity inform ation. However, toxicity inform ation is needed to confirm this assum p tion.
Haz a rd Concerns

. Acute toxicity for three m e m b ers of this category are available and all have been shown to be m oderately toxic to aquatic organism s (i.e., acute toxicity values between 1 and 100 m g /L): benzoyl chloride, fish 96-h LC50 = 35.0 m g /L, an arom atic dicarboxyl dichloride, fish 96-h LC50 = 6.2 m g /L, and benzene sulfochloride, fish 48-h LC50 = 3.0 m g /L. All of these tests have been done with the static m e thod using nom inal concentrations. It is unclear just how acid chlorides are toxic to aqua tic organism s. It is known that acid chlorides hydrolyze to the carboxylic/sulfonic acid and HCl. It is not known if the toxic effect is the result of (1) absorption of the acid chloride and hydr olysis within the m e m b rane, or (2) the HCl produced from the hydrolysis. It is known th at the carboxylic/sulfonic-acid hydrolysis products are of low toxicity. Boundaries

. There are no known lower boundaries. The upper boundaries will be based on Kow and MW when enough inform ation is obtained. In general, when the log K owvalue is < 8, the environm ental base set of tests will be requested f o r aquatic releases and the terrestrial base set of tests will be recomm e nded for terrestrial exposures. Wh en the log K owis > 8, testing will be requested until enough inform ation is obtained to determ ine whether these com pounds will have no toxic effects at saturation. Generally, m e m b ers of this category will have MW s of less than 1000 but testing of m e m b ers with a MW > 1000 m ay be requested to confirm whether acid chlorides have to be absorbed to be toxic. General Testing Strategy. The testing strategy f o r acid chlorides will consist of two steps. (1) Hydrolysis as a function of pH at 25 °C (40 CFR 796.3500) will be recomm e nded. Depending on the outcom e of this environm ental fate testing and reassessm ent, (2) the aquatic base set of environm ental toxicity tests will be recomm e nded for aquatic exposures with the fish acute toxicity test done once or twice. Chronic toxicity testing for aquatic organism s include: the fish early life state toxicity test, the daphnid partial life cycle toxicity test and the algal toxicity test. The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm acute toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial exposures. Chronic toxicity te sting for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay. October, 1990

1Category: Acid Dyes and Amphoteric Dyes Environm ental Toxicity Definition. Organic dyes are divided into four cl asses depending on the type of electronic charge of the dye: nonionic (neutral dyes); anionic (negative charge or acid dyes); cationic (positive charge) dyes; and am photeric (m ixture of positive and negative charges on sam e m o lecule) dyes. Nonionic or neutral dyes are assessed as neutral organic chem icals for which there is a separate category description, and cationic dyes also have a separate category description. Am photeric dyes are assessed eith er as cationic or anionic dyes depending on dom inant net charge.
Haz a rd Concerns. Analysis of over 200 acid dyes (Auer et al. 1990, Nabholz 1990, Sigm an et al. 1983, Tonogai et al. 1979, Little and Lam b 1972, ADMI 1974) have indicated that som e m onoacid and diacid dyes can show m oderate to high toxicity (i.e., acute values < 100 m g /L and < 1 m g /L) to fish and aquatic organism s. Dyes with three or m o re acid groups showed low toxicity (i.e., acute values > 100 m g /L) towards fish and invertebrates. Som e m e tal chelated dyes, i.e., Al, Co, Cr, Fe, have shown m oderate toxicity towards fish and daphnids and the toxicity has not been explained by the residual free (un-chelated) m e tal ion in the dye product. All acid dyes showed m oderate toxicity towards green algae. Analysis of available data (Auer et al. 1990, Nabholz 1990) has suggested that effects to algae were not the result of direct toxicity but represented an indirect effect due to shad ing. Senior regulatory decision-m akers in OPPT (then OTS) decided in 1988 that the risk to alg ae from indirect (shading) toxicity was not an unreasonable risk for two m a in reasons: (1) algae grew quickly as soon as the dye was diluted, and (2) the release of colored effluents in the U. S. generally results in imm e diate com p laints by citizens to their local authorities, e.g., county and state governm e nts. The rapid response by the public generally results in quick regulatory action by local officials. Since there is no SAR for acid dyes (Auer et al. 1990), hazard profiles for m onoacid dyes are developed using m easured data or data for the nearest analog(s). Boundaries. Acids dyes m u st have som e water solubility and m o lecular weights generally need to be near or below 1000. General Testing Strategy. I. Release to Aquatic Ecosystem s : Tier 1. Because of the above boundary conditions, and the need to assess hum an exposure as well as environm ental toxicity, if there is insufficient knowledge about the water solubility of the dye, then it should be m easured (40 CFR §796.1860). The fish and daphnid acute toxicity tests f r om the aquatic base set of environm ental toxicity tests will be recomm e nded for aquatic exposures. The acute toxicity tests for fish (40 CFR §797.1400) and daphnids (40 CFR §797.1300) will be done using the flow-through m e thod with m easured concentrations, and effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations. If there is no significant risk from the PMN after the results of the fish and daphnid acute

2toxicity tests have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Direct and Indirect Photolysis Screening Test (40 CFR 796.3765). If t1⁄2 < 2 days, go to Tier 3; if t1⁄2 > 2 days, go to Tier 4. Tier 3a. If t1⁄2 < 2 days and photolysis products are known and/or identified, then assess photolysis products for environm ental hazards. Tier 3b. If t1⁄2 < 2 days and photolysis products are not known and/or identifiable, then prepare a stock solution of PMN using the standard hum ic-containing solution described in the direct and indirect photolysis screening test [40 CFR 796.3765 (b)(2) and (c)(2)], expose to sunlight for at least 6 half-lives (t1⁄2), and test photolysis products for toxicity with m o st sensitive species from environm ental base set. Tier 4. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (40 CFR 797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; and statistical analysis of effective concentrations at days 7, 14, 21, and 28. Daphnid chronic toxicity testing ( 40 CFR 797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21. Aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Aerobic Aquatic Biodegradation Modified Sturm Test Closed Bottle Test Modified OECD Screening Test Modified MITI Test (I) Modified AFNOR Test
II. Release to Terrestrial Ecosystem s : 40 CFR 796.3100 40 CFR 796.3260 40 CFR 796.3200 40 CFR 796.3240 40 CFR 796.3220 40 CFR 796.3180

The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded f o r terrestrial exposures. Chronic toxicity testing fo r terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay.

3References . Am erican Dye Manufacturers Institute, Inc. (ADMI), 1974, "Dyes and the Environm ent: Reports on Selected Dyes and Their Effects," Vol. II, ADMI, New York. Auer, C.M., Nabholz, J.V. and Baetcke, K.P., 1990, "Mode of Action and the Assessm ent of Chem ical Hazards in the Presence of Lim ited Data: Use of Structure-Activity Relationships (SAR) under TSCA, Section 5," Environm ental Health Perspectives , Vol. 87, pp. 183-197. Little, L.W

. and Lam b J.C., III, 1972, "Acute Toxicity of 46 Selected Dyes to the Fathead Minnow, Pim e phales prom elas ,"Final Report to the Am erican Dye Manufacturers Institute,Inc., UNC W a stewater Research Ce nter, Departm e nt of Environm ental Sciences and Engineering, School of Public Health, Un iversity of North Carolina, Chapel Hill, NC. Nabholz, J.V., 1990, "The OTS PMN ECOTOX Data Base: a Confidential Business Inform ation (CBI) Collection of Environm ental Toxic ity Data from New Chem ical Subm issions Under Sec. 5," Unpublished, Environm ental EffectsBranch, Health and Environm ental Review Division (TS-796),U. S. Environm ental Protection Agency, W a shington, DC. Sigm an, C.C., Helm s, C.T., Papa, P.A., Atkinson, D.L., Doeltz, M.K. and W i nship-Ball, A., Jan. 1983, "Anthraquinone Dyes and Related Chem icals: Review and Assessm ent of Potential Environm ental and Health Aspects," Final Report to the Dyes Environm ental and Toxicology Organization, Inc., SRI International, Menlo Park, CA. Tonogai, Y., Ito, Y., Iwaida, M., Tati, M., Ose, Y. and Sato, T., 1979, "Studies on the Toxicity of Coal-Tar Dyes. II. Exam ination of the Biological Reaction of Coal-Tar Dyes to Vital Body," The Journal of Toxicological Sciences , Vol. 4, pp. 211-219. July, 1991; revised June, 1994

4Category: Acrylamides Hum an Health and Environm ental Toxicity Definition. Any new chem ical with the following structure is considered to be a m e m b er of the category:

A typical acrylam ide in the new chem ical program is used as a m onom er and has a m o lecular weight of < 500. The acrylam ides of greatest concern are those with a labile substituent, e.g., m e thylol acrylam ides, that m ay release acrylam ide per se under m e tabolic conditions. For toxicity to aquatic organism s, there is a concern f o r all substituted acrylam ides with m o lecular weights of <1000 and log Kow of <8.0.
Haz a rd Concerns. Based on analogy to acrylam ide per se , m e m b ers of the class are considered potential carcinogens, heritable m utagens, reproductive and developm ental toxicants, and toxic to aquatic organism s. Acrylam i des are also potential neurotoxins based on data for a num ber of low m o lecular weight acrylam ides. Boundaries. Structures with an acrylam ide equivalent weight of > 5,000 are presum ed not to pose a hazard under any conditions. Typically, concerns are confined to those species with m o lecular weights <1,000 whenever inhalation (or environm ental) exposure is expected, and to species <500 when derm al, but not inhalation, exposure to hum ans is expected. Occupational Exposure Controls. Because acrylam ides are expected to be absorbed via the derm al route, glove perm eation testing conducted in accordance with standard ASTM testing protocols m ay be required, depending upon estim ated workplace exposures and the hazard identif ied f o r the particular acrylam ide subm itted as a PMN.

5General Testing Strategy . Tier 1. To properly assess hum an and environm ental toxicity or exposure, certain physicalchem ical or environm ental fate properties need to be m easured: water solubility (40 CFR §796.1860) Kow (40 CFR §796.1570 or 40 CFR §796.1550) vapor pressure (40 CFR §796.1950) m e lting point-m elting range (OECD 102 or OPPTS 830.7200) Aerobic biodegradability can be determ ined using one of the following test guidelines, listed in approxim a te order of preference: Aerobic aquatic biodegradation Modified Sturm test Closed bottle test Modified OECD screening test Modified MITI test (I) Modified AFNOR test 40 CFR 796.3100 40 CFR 796.3260 40 CFR 796.3200 40 CFR 796.3240 40 CFR 796.3220 40 CFR 796.3180 The physical state and electronic charge of the PMN substance should also be reported. Tier 2. EPA considers the following tests to be the m o st appropriate for acrylam ides found to pose an unreasonable risk to hum an health: - 90-day subchronic toxicity (40 CFR §798.2650) with functional observational battery (40 CFR §798.6050) and neuropathology (40 CFR §798.6400). - A rodent dom inant lethal assay (40 CFR §798.5450). If positive, a rodent heritable translocation test (40 CFR §798.5460) would be the appropriate followup test. - A 2-year carcinogenicity test (40 CFR §798.3300) in rats and m i ce. To address environm ental toxicity concerns, the following testing is recomm e nded, for acrylam ides with log Kow <5 : acute aquatic toxicity testing in algae, daphnid, and fish. The acute toxicity tests for fish (40 CFR 797.1400) and daphnids (40 CFR 797.1300) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; m easured TOC of dilution water in the control <2 m g TOC/L; the highest treatm e nt concentration on a m ean m easured-basis should equal the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. The algal toxicity testing (40 CFR 797.1050), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and

696 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; and the highest treatm e nt concentration on a m ean m easured-basis equal to the aqueous solubility lim it. Solvent can be used to assist the PMN substance to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN substance beyond its aqueous solubility lim it. For acrylam ides with log Kow >5 and <8 , aquatic toxicity testing should be for chronic effects only: (1) fish chronic toxicity testing, i.e ., fish early life stage (ELS) toxicity testing with rainbow trout (40 CFR 797.1600), with flow-thr ough m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; and statistical analysis of effective concentrations at days 30, 45, 60, 75, and 90; (2) daphnid chronic toxicity testing (40 CF R 797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m e an m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; and (3) algal toxicity testing (40 CFR 797.1050), with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; and the highest treatm ent concentration on a m ean m easured-basis equal to the aqueous solubility lim it. Solvent can be used to assist the PMN substance to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN substance beyond its aqueous solubility lim it.
Novem b er 1990; revised July 1993; revised October 1995

7Category: Acrylates/Methacrylates Environm ental Toxicity (Hum an Health, case-by-case) Definition. Any m o lecular structure containing one or m o re of the following reactive groups is considered to be a m e m b er of the class:
O 5 - acrylate, CH 2 =CH-C-O-

O 5 - methacrylate, CH 2 =C-C-O­ | CH 3

Haz a rd Concerns.
Hum an Health: As a result of testing conducted under an agreem ent between the Agency and the Specialty Acrylates and Methacrylates (SAM) Panel of the Chem ical Manufacturers Association (CMA), EPA no longer controls new chem ical acrylates or m e thacrylates as a category based on health concerns. However, if a new acrylate is structurally sim ilar to a substance for which EPA has positive toxicity data, EPA m ay regulate that substance under TSCA section 5(e) based on its potential unreasonable risk. This will be done on a case-by-case basis and is expected to effectively elim inate m o st regulation of acrylates, especially higher m o lecular weight and polym eric substances. Despite the fact that EPA no longer expects to m ake a potential unreasonable risk to hum an health finding for m any of the new acrylates, EPA still recomm e nds the use of engineering controls or personal protective equipm ent to reduce exposures in the workplace in recognition of their potential as irritants and sensitizers. Environm ental Toxicity: The ecotoxicity of acr ylates and m e thacrylates is a function of the octanol-water partition coef f i cient. They exhibit sim p le narcosis at log P' s >5, but display excess toxicity at lower log P' s. The toxicity of acrylates and m e thacrylates can be predicted by a QSAR (quantitative structure-activity relationship), although there are som e m e m b ers of the class such as allyl m e thacrylate that are significantly m o re toxic than predicted by the QSAR. Boundaries. Typically, environm ental toxicity concerns are confined to those species with m o lecular weights <1,000. Acute and chronic toxi city is possible at log P’s <5, and chronic toxicity is possible at log P’s <8. General Testing Strategy Tier 1. To properly assess environm ental toxicity or exposure, certain physical-chem i cal or environm ental fate properties need to be m easured:
8water solubility (40 CFR §796.1860) log Kow (40 CFR §796.1570 or 40 CFR §796.1550) vapor pressure (40 CFR §796.1950) m e lting point-m elting range (OECD 102 or OPPTS 830.7200) aquatic biodegradation (40 CFR §796.3100). The physical state and electronic charge of the PMN substance should also be reported. Tier 2. To address environm ental toxicity con cerns, the following testing is recomm e nded: acute aquatic toxicity testing in algae, daphnid, and fish (all tests using m e asured concentrations; algae: static m e thod, daphnid and fish: flow-through m e thod). Septem ber, 1988; Revised July, 1993 and January, 1997.

9Category: Aldehydes Environm ental Toxicity Aldehydes are a class of organic compounds characterized by the functional group RC(=O)-H. Aldehydes are ionizable in water and exhibit excess aquatic toxicity in addition to narcosis. Polyaldehydes are m o re toxic than m onoaldehydes to aquatic organism s and acrylic/vinyl/allylic aldehydes , e.g., acrolein, are m o re toxic than polyaldehydes. Vinyl/allylic/acrylic aldehydes are a class of organic com pounds characterized by both the aldehyde, R-C(=O)-H, and vinyl, H 2 C=C-R, or allylic, R-C=C-R, functional groups. Allylic and vinyl aldehydes, e.g., acrolein, C=CC(=O), exhibit excess toxicity. It is assum e d that aldehydes need to be absorbed to be toxic, therefore, aldehydes with MW >1000 will be excluded from this category. Acute toxicity for aldehydes is known to be lim ited by the octanol/water partition coef f i cient (Kow ). Above a log K owvalue of >6.0, aldehydes show no effects at saturation during 96-h exposures to fish. Aldehydes which are solids at room tem p erature m ay show no toxicity at saturation at lower K owvalues depending on the m e lting point, i.e., the higher the m e lting point at a given K ow , the greater the likelihood that no toxicity will be observed at saturation. For solids, the no effects at saturation has to be determ ined on a case-by-case basis. There are no m easured upper Kowlim its f o r chronic toxicity at this tim e, but it m ay not be m u ch above a log K ow= 8. Future testing will determ ine K ow lim its.
Haz a rd Concerns : The toxicity for aldehydes has been determ ined through SAR Analysis: SARs for monoaldehydes : log fish 96-h LC50 (m oles/L) = -0.449 log K ow-3.314 where n=54, R2 =0.53, CLOGP<6, MW <1000; log daphnid 48-h LC50 (m illim oles/L) = -0.059 -0.608 log Kow where n=5, R2 =1.0, CLOGP<6, MW <1000; log green algal 96-h EC50 (mm o l/L) = 0.994 -0.812 log K ow(CLOGP) where n=2, R2 =1.0, CLOGP<6.4, MW <1000; log fish ChV (m illim oles/L) = -0.810 -0.680 log K ow(CLOGP) where n=3, R2 =0.97, CLOGP<8, MW <1000; log daphnid ChV (m illim oles/L) = -1.090 -0.576 log K ow(CLOGP) where n=2, R2 =1.0, CLOGP<8, MW <1000; and log algal ChV (mm o l/L) = 0.053 -0.644 log K ow(CLOGP) where n=5, R2 =0.99, CLOGP<8, MW <1000. SARs for polyaldehydes :

10log fish 96-h LC50 (m illim oles/L) = -1.176 -0.371 log K ow(CLOGP) where n=2, R2 =1.0, CLOGP<6, MW <1000; log fish 96-h LC50 (m illim oles/L) = -1.021 -0.396 log K ow(SRC) where n=2, R2 =1.0, SRC<6, MW <1000; log daphnid 48-h LC50 (mm o l/L) = -1.059 -0.440 log K ow(CLOGP) where n=2, R2 =1.0, CLOGP<6, MW <1000; log daphnid 48-h LC50 (mm o l/L) = -0.875 -0.471 log K ow(SRC) where n=2, R2 =1.0, SRC<6, MW <1000; log green algal 96-h EC50 (mm o l/L)= -1.772 -0.364 log K ow(CLOGP) where n=3, R2 =0.99, CLOGP<6.4, MW <1000; log green algal 96-h EC50 (mm o l/L)= -1.620 -0.388 log K ow(SRC) where n=3, R2 =0.99, CLOGP<6.4, MW <1000; log fish ChV (m illim oles/L) = -2.298 -0.488 log K ow(CLOGP) where n=2, R2 =1.0, CLOGP<8, MW <1000; log fish ChV (m illim oles/L) = -2.092 -0.513 log K ow(SRC) where n=2, R2 =1.0, CLOGP<8, MW <1000; log daphnid ChV (m illim oles/L) = -1.798 -0.488 log K ow(CLOGP) where n=2, R2 =1.0, CLOGP<8, MW <1000; log daphnid ChV (m illim oles/L) = -1.592 -0.513 log K ow(SRC) where n=2, R2 =1.0, CLOGP<8, MW <1000; log algal ChV (m illim oles/L) = -2.313 -0.348 log Kow(CLOGP) where n=2, R2 =1.0, CLOGP<8, MW <1000; and log algal ChV (m illim oles/L) = -2.166 -0.367 log Kow(SRC) where n=2, R2 =1.0, CLOGP<8, MW <1000 SARs for vinyl/allylic/acrylic aldehydes : log fish 96-h LC50 (mm o les/L) = -3.502 +0.017 log K ow(CLOGP) where n=3, R2 =0.25, CLOGP<6, MW <1000; and log fish ChV (m illim oles/L) = -4.377 -0.228 log K ow(CLOGP) where n=2, R2 =1.0, CLOGP<8, MW <1000.

11Environmental Fate :

Boundaries : MW <1000. Log K ow <6.4 for toxicity to green algae as a 96-h EC50; and log K owassum e d to be <8.0 for chronic toxicity to aquatic organism s, but could be higher. <6.0 for acute toxicity to fish and aquatic invertebrates; log K ow General Testing Strategy : I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity tests for fish (CFR §797.1400) and daphnids (CFR §797.1300) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; m easured TOC of dilution water in the control; ideally, m e an m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; and solvent can be used to assist the aldehyde to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the aldehyde beyond its aqueous solubility lim it. Stock solutions in water should be adjusted to pH near 7.0. The algal toxicity testing (CFR §797.1050), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; and solvent can be used to assist the aldehyde to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the aldehyde beyond its aqueous solubility lim it. Stock solutions in water should be adjusted to pH near 7.0. If there is no significant risk from the aldehyde after the results of the environm ental base set have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Aerobic Aquatic Biodegradation 40 CFR 796.3100 Modified Sturm Test 40 CFR 796.3260 Closed Bottle Test 40 CFR 796.3200 Modified OECD Screening Test 40 CFR 796.3240 Modified MITI Test (I) 40 CFR 796.3220 Modified AFNOR Test 40 CFR 796.3180

12If there is no significant risk from the aldehyde after the results of biodegradation testing have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 3. Tier 3. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (CFR §797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; m easured TOC of dilution water in the control; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test because the 7-d ELS toxicity test m ay underestim ate chronic toxicity m easured by the 28-d ELS toxicitytest when the Chronic Values are com p ared. Stock solutions in water should be adjusted to pH near 7.0. Daphnid chronic toxicity testing (C FR §797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; m easured TOC of dilution water in the control; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test because the fish 7-d ELS toxicity test m ay underestim ate chronic toxicity m easured by the fish 28-d ELS toxicity test when the chronic values are com p ared.
II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial exposures. Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay. July, 1996

13Category: Aliphatic Amines Environm ental Toxicity This category includes prim ary am ines, secondary am ines and tertiary am ines; or m onoalkyl am ines, dialkly am ines and trialkyl am ines, respectively. This group includes alkanes, alkenes and alkynes; substitutions on carbon (alkyl) chains m ay include but not be lim ited to halogens and hydroxyls; insertions in alkyl chain m ay include but not be lim ited to ethoxys, propoxys, ethers, sulfides, disulfides and polysulfides; am ine oxides are also included in this category; fatty polyam i nes (e.g., diam ines, triam i nes, tetraam ines, pentam ines, etc) are also included; am ines m ay either be un-ionized (free) or i onized; and strong ion pairs m ay also be included.

Haz a rd Concerns: Mem b ers of this category can be highly toxic to all groups of freshwater organism s (i.e., fish, aquatic invertebrates and greenalgae). Toxicity is related to the length of the hydrophobic carbon chains: the longer (or greater the num ber of carbons) the chain the m o re toxic to aquatic organism s when the num ber of am ines is constant; and the greater the num ber of am ines, the greater the toxicity given a constant carbon chain length. Sm all aliphatic am ines are m o re toxic to algae than fish and invertebrates; higher m o lecular weight am ines are equally toxic to all aquatic organism s. Sm all aliphatic am ines which are un-ionized are m o re toxic to fish than when they are ionized; toxicity to algae appears to be unaffected by ionization. Strong ion pairs are generally m u ch less toxic to fish and invertebrates because of solubility lim itations, but rem a in highly toxic to green algae. The toxicity of each am ine will be predicted using structure-activity relationships (SARs) and analogs contained in a generic standard environm ental hazard review for aliphatic am ines. Boundaries: There are no lower boundaries and the upper boundary is unknown at this tim e. It is known that a C13-NH3 Cl is still toxic to fish at less than 1 m g /L. An upper boundary for carbon chain length will probably be about 20 carbons but m o re inform ation is needed at this tim e. Generally, m e m b ers of this category will have m o lecular weights less than 1000.

14General Testing Strategy Tier 1. The base set of environm ental toxicity tests in clean dilution water and two fish acute toxicity tests done in the presence of hum ic acid (i.e., TOC) will be recomm e nded. These test will be done under static m e thods with nom inal concentrations. Sm aller am ines (e.g., Schiff bases) are expected to dem onstrate less m itigation by hum ic acid (larger log P, greater m itigation; increase log P correlates to increased MW ). If hydrolysis 1/2 life is less than one hour, test the hydrolysis product; if it is greater than one hour, test parent m a terial. Tier 2. If TOC significantly reduces the toxicity to water columnspecies, then toxicity testing f o r toxicity to benthic organism s (i.e., organism s that ingest sedim e nt) will be recomm e nded; however, if TOC does not significantly reduce the toxicity to water column species, then chronic toxicity with f i sh and aquatic invertebrates will be recomm e nded as well as aerobic biodegradation testing. Aerobic biodegradability can be determ ined using one of the following test guidelines, listed in approxim a te order of preference: Aerobic aquatic biodegradation Modified Sturm test Closed bottle test Modified OECD screening test Modified MITI test (I) Modified AFNOR test
Septem ber 1988; revised Novem b er 1995. 40 CFR 796.3100 40 CFR 796.3260 40 CFR 796.3200 40 CFR 796.3240 40 CFR 796.3220 40 CFR 796.3180

15Category: Alkoxysilanes Hum an Health Environm ental Toxicity Definition. Any m o lecular structure containing one or m o re of the following reactive groups is considered to be a m e m b er of the category. | | -C-O-Si­ | | The "typical" new chem ical of concern is a polym er with a substantial fraction of species with m o lecular weights <1000 and pendant trim ethoxy- or triethoxysilane groups.
Haz a rd Concerns. Health- Concern for lung toxicity from inhalation of vapors or aerosols is based on data for a num ber of low-m o lecular-weight alkoxysilanes. Trim ethoxysilane (TMS) is clearly the m o st toxic m e m b er of the class causing irreversible lung effects at low doses, but the Agency does not consider it appropriate to use TMS as a regulatory benchm ark for all alkoxysilanes. For trim ethoxysilane m onom ers and polym ers with a low trim ethoxysilyl equivalent weight, a NOAEL of 10 ppm (about 11 m g /kg/day) based on a 90-day study with vinyltrim ethoxysilane in m onkeys is deem ed an appropriate generic benchm ark. Alkoxysilanes in which the alkyl substituent is not a m e thyl group do not appear to be as toxic as m e thoxysilanes. The New Chem icals Program currently uses a generic benchm ark NOAEL of 75 m g /kg/day, based on a 90-day inha lation study with tri(isopropenoxy)silane, for alkoxysilanes other than m e thoxysilanes. Ecotoxicity- Alkoxysilanes are highly toxic to algae and m oderately toxic to aquatic invertebrates. For exam ple, the daphnid 48-hr LC 50 for dim e thyldiethoxysilane is 1.25 m g /L, and the 15-day algal EC 95's for vinyltriethoxysilane, tetraethoxy-silane, and trifluoropropenyl(m ethyl)diethoxysilane are all approxim a tely 10 

### : g/L. Boundaries. Methoxy- and ethoxysilanes are presum ed not to pose a hazard under any conditions if the equivalent weight is 

### $ 5,000 and no m o re than 25% of species have m o lecular weights less than 1,000 and no m o re than 10% of species have m o lecular weights less than 500. For alkoxysilanes with alkyl substituents larger than propyl groups, the equivalent weight cutoff is 1,000. The degree of concern depends on the relative abundance of lower m o lecular weight species, but there is no m o lecular weight threshold above which there would be no concern. To better define the boundaries of the category, EPA seeks testing on a lim ited num ber of alkoxysilanes that focuses on (1) the relationship between m o lecular weight (or alkoxysilyl equivalent weight) and inhalation toxicity and (2) the im portance of increasing alkoxy chain length in lim iting toxicity.

16General Testing Strategy The Agency recomm e nds the following testing as appropriate to address health and environm ental toxicity concerns for this category: 1. 90-day subchronic test in rodents by the inhalation route (40 CFR 798.2650). 2. Hydrolysis testing (40 CFR 796.3500). If t 1/2 is less than one hour, base set ecotoxicity testing (see "3," below) is conducted with the hydrolysis products only. If t 1/2 is greater than one hour, base set ecotoxicity testing is conducted with the parent m a terial; the PMN subm itter has the option of also testing with the hydrolysis products. 3. Base-set ecotoxicity testing to include fish (40 CFR 797.1400) using the static m e thod, daphnids (40 CFR 797.1300) using the static m e thod and algae (40 CFR 797.1050) using the static m e thod, all nom inal concentrations. Direct dilution of the test alkoxysilane and organism s is added within 10 m i nutes. The static-renewal m e thod is used for fish and daphnid test, plus an additional fish test using aged stock solution. Results of the acute ecotoxicity testing m ay trigger chronic fish (40 CFR 797.1600) and daphnid (40 CFR 797.1350) testing. 4. Physical-chem i cal or environm ental fate testing including, as appropriate, m e lting point (40 CFR 796.1300) or boiling point (40 CF R 796.1220), water solubility (40 CFR 796.1840 or 796.1860), log K ow(40 CFR 796.1550, 796.1570 or 796.1720), vapor pressure (40 CFR 796.1950), direct photolysis and indirect phot olysis (40 CFR 796.3765). Need for water solubility, log K ow , and photolysis testing determ ined by outcom e of above hydrolysis testing. Septem ber, 1988; revised June, 1994

17Category: Aluminum Compounds Risk Management Statement. Greatest concern is for soluble form s of alum inum (Al). If water solubility is greater than 1 part per billion (1 ppb), the Agency will prohibit releases of the PMN substance to water pending the subm ission of environm ental toxicity testing. Definition. This category includes inorganic salts of Al, com p lexes between Al and organic acids or chelates of Al by polyanionic m onom er s, and organoAl compounds, i.e., Al covalentlybonded with carbon. For exam ple, som e inorganic Al salts include: Al hydroxide, Al chloride, Al fluoride, Al nitrate, Al phosphate, and Al su lfate. Not included in this category are dyes com p lexed with Al (see dye categories addressed elsewhere within this “TSCA New Chem icals Program Chem ical Categories” docum ent).
Haz a rd Concerns. Soluble salts of Al are known to be highly toxic to green algae and m oderately toxic to fish and aquatic invertebrates at pH values between 6.5 to 9.0 and in term s of soluble Al in m g Al/L. Toxicity inform ation ar e available for Al chloride, Al sulfate, and Na alum inate. The Office of W a ter (USEPA, 1988, EPA440/5-86-008) cited Seipt et al. (1984, W a ter Air Soil Pollut. 23:81-95) who concluded that "the sim p le hydroxides (Al(OH) 2+ and Al(OH) 2 + ) are regarded as the m o st dangerous form s while organically bound Al and polym eric form s are less toxic or essentially harm less." The Office of W a ter (USEPA, 1988) also concluded that solutions of Al in water approachchem ical equilibrium rather slowly and that Al can form strong com p lexes with fulvic and hum ic acids. The toxicity profile for soluble Al salts , listed below, is based on (1) available m e asured (M) toxicity data, (2) m g Al/L (ppm Al), (3) pH between 6.5 and 9.0, and (4) m oderate hardness (about 150.0 m g /L as CaCO 3 ). fish (FHM) 96-h LC50 = 35.0 M pH7.3 H220 fish (RT) 96-h LC50 = 8.6 M pH7.5 H47 fish (RT) 96-h LC50 = 7.4 M pH6.6 H47 fish (RT) 96-h LC50 = 14.6 M pH7.3 H47 m ean RT 96-h LC50 = 10.0 P n3 fish (BT) 96-h LC50 = 3.6 M pH6.5 H? m ean fish 96-h LC50 = 11.0 P n3 daphnid (Cd) 48-h LC50 daphnid (Cd) 48-h LC50 m ean Cd 48-h LC50 daphnid (Dm ) 48-h LC50 daphnid (Dm ) 48-h LC50 m ean Dm 48-h LC50 m ean daphnid 48-h LC50 OW FW Acute W Q C = 1.9 = 3.7 = 2.7 = 3.9 = 38.0 = 12.0 = 5.7

= 1.5 M pH7.4 H50 M pH7.7 H47 P n2 M pH7.0 H45 M pH7.1 H220 P n2 P n2

18---------------------------------------------------

--------------------------------------------------- green algal 96-h EC50 green algal 96-h EC50 m ean algal 96-h EC50 fish (FHM) ChV fish (FHM) ACR daphnid (Dm ) ChV daphnid (Dm ) ChV m ean Dm ChV Dm ACR daphnid (Cd) ChV Cd ACR m ean daphnid ChV m ean daphnid ACR OW FW Chronic W Q C algal ChV Biological Fate = = =

= =

= = = = = = = =

=

= 0.570 M pH7.6 H15 0.460 M pH8.2 H15 0.510 P n2 3.3 M pH7.7 H220 11.0 M 0.742 M pH8.3 H220 0.320 M pH7.7 H45 0.490 P n2 24.0 M

1.9 M pH7.2 H50 1.4 M 0.970 P n2 5.9 M 0.087 0.100 P EC50÷4

Fish (BT-eyed em bryo) 30-d BCF (wb) = 50.0 M pH7.2 H242 Fish (BT-fry) 30-d BCF (wb) = 136.0 M pH7.2 H242 m ean fish BCF = 93.0 P n2 The toxicity profile for soluble inorganic com p lexes of Al can be predicted via MW adjustm e nt of

the toxicity f o r Al to the toxicity of the com p lex. Ca Al hydroxy phosphites [141 728-04-3] Ca xAl 2 (OH) 2 (x+3-y) (HPO 3 ) y 

### @ mH 2 O with x = 2 to 12, (2x+5)÷2 >y>0, and m = 0 to 12; typical com position = 31% Ca, 9.5% Al, & 8.0% P; solid with m p >250 °C (dec); S = 680 m g /L; pH 11.5; Concentration (m g/L) Effect

---------------------------- Com p lex Al Notes fish 96-h LC50 120.0 11.0 P fish 96-h LC50 409.0 6.0 M daphnid 48-h LC50 60.0 5.7 P daphnid 48-h LC50 24.0 nm M green algal 96-h EC50 5.4 0.510 P fish ChV 35.0 3.3 P

19---------------------------------------------------

--------------------------------------------------------------

--------------------------------------------------------------

-------------------------------------------------------------- daphnid ChV 10.0 0.970 P algal ChV 1.0 0.100 P

Na Al fluoride [15 096-52-3] sodium fluoalum i nate; sodium fluoride alum inum ; cryolite; kryolith; M12,2673; Na 3 Al F 6 ; MW 210; com position: 13% Al; solid with m p 1000 °C; S = 610 m g /L with pH 6.2; used as an insecticide since 1929; Concentration (m g/L) Effect

------------------------------------------- Com p lex A l N otes fish 96-h LC50 85.0 11.0 P fish 96-h LC50 21.0 2.8 M daphnid 48-h LC50 44.0 5.7 P green algal 96-h EC50 3.9 0.510 P fish ChV 25.0 3.3 P daphnid ChV 7.5 0.970 P algal ChV 0.770 0.100 P

The toxicity profile for chelates of Al with polyanionic m onom ers can be predicted via MW adjustm e nt of the toxicity for Al to the toxicity of the chelate. The toxicity profile for organoAl com pounds are developed for only the hydrolysis product(s) of Al. OrganoAl com pounds are unstable in air and water. MethylAl and ethylAl are pyrophoric and dodecylAl slowly hydrolyses in water. Boundaries. The toxicity of Al com pounds depends on the their water solubility, the bioavailability of Al, and their stability. The m o st im portant property determ ining the toxicity of Al com pounds is water solubility. W a ter solubility cannot be predicted accurately and has to be m easured. Molecular weight (MW ) is only im portant when Al com p lexes are water soluble and stable. Stable com p lexes of Al with MW s > 1000 are not expected to be absorbed by aquatic organism s and Al is not expected to be bioavailable even if they are water soluble. Therefore, only unstable Al com pounds with MW s < 1000 are expected to be toxic. General Testing Strategy Tier 1. Fate testing

. Physical state (OPPTS 830.6303) and corresponding property, i.e., m e lting point-m elting point range (OPPTS 830.7200) or boiling point-boiling point range (OPPTS 830.7220), water solubility (OPPTS 830.7840 or 7860), octanol/water partition coefficient (Kow ) (OPPTS 830.7550 or 7570), and vapor pressure (OPPTS 830.7950); and/or

20Acute environm ental toxicity testing

. The aquatic base set of environm ental toxicity tests will be recomm e nded for aquatic exposures: fish acute t oxicity, daphnid acute toxicity, and green algal toxicity. The acute toxicity tests for fish (40 CFR §797.1400 or OPPTS 850.1075) and daphnids (40 CFR §797.1300 or OPPTS 850.1010) will be done using the static m e thod; effective concentrations will be based on 100% active ingredients (ai) and nom inal concentrations; the total organic carbon (TOC) concentration of dilution water in the control m u st be less than 2.0 m g TOC/L; TOC m u st be m easured in the control just prior to the start of the test; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested com pound; solvent can not be used; and hardness of dilution water has to be less than 180.0 m g /L as CaCO 3 . The algal toxicity testing (40 CFR §797.1050 or OPPTS 850.5400), should be done with the static m e thod; effective concentrations based on 100% ai and nom inal concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with no m o re than 0.300 m g /L EDTA as a final concentration; the TOC of the test/growth m e dium should be less than 2.0 m g TOC/L; TOC should be m easured just prior to the start of the test; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested com pound; and solvent can not used;

. In addition fish-toxicity-m itigation testing (OPPTS 850.1085) with known am ounts of hum ic acid (HA) added to dilution water, i.e., 20 m g HA/L and 10 m g HA/L, will be recomm e nded. If there is no significant risk from the Al com pound after the results of tier one testing set have been integrated into the risk assessm ent, then no f urther testing will be recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Fate testing

. Aerobic biodegradation, i.e., ready biodegradation (OPPTS 835.3110) or sealed-vessel test (OPPTS 835.3120); and/or Fish chronic toxicity testing , i.e., fish early life stage (ELS) toxicity testing (40 CFR §797.1600 or OPPTS 850.1400), with the flow-through m e thod; effective concentrations based on 100% ai and m ean m easured concentrations of soluble Al; statistical analysis of effective concentrations at days 7, 14, 21, and 28; the TOC of dilution water in the control should be less than 2.0 m g TOC/L; TOC should be m easured in the controls just prior to and during the test; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested com pound; solvent can not be used; and hardness of dilution water has to be less than 180.0 m g /L as CaCO 3 ; and Daphnid chronic toxicity testing (40 CFR §797.1330 or OPPTS 850.1300), with the flowthrough m e thod; effective concentrations based on 100% ai and m ean m easured concentrations of soluble Al; statistical analysis of effectiv e concentrations at days 7, 14, and 21; the TOC of dilution water in the control should not exceed 2.0 m g TOC/L; TOC m u st be m e asured in the controls just prior to and during the test; the highest treatm e nt concentration on a nom inal-basis

21should not exceed the aqueous solubility lim it of the tested com pound; solvent can not be used; and hardness of dilution water has to be less than 180.0 m g /L as CaCO 3 . II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests will be recomm e nded for terrestrial exposures. The terrestrial base set includes: the early seeding growth test (40 CFR 797.2800 or OPPTS 850.4230), the earthworm toxicity test (40 CFR.795.150 or OPPTS 850.6200), the soil m i crobial comm unity bioassay (40 CFR 797.3700 or OPPTS 850.5100), and the avian acute oral t oxicity test (40 CFR 797.2175 or OPPTS 850.2100). Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test (OPPTS 850.xxxx), the plant uptake test (40 CFR 797.2850 or OPPTS 850.4800), and the avian reproduction test (OPPTS 850.2300). Abbreviations. ACR10 = acute-to-chronic ratio = 10; ai = active ingredients; BCF = bioconcentration factor; BT = brook trout; Cd = Ceriodaphnia ; ChV = Chronic value; Dm = Daphnia magna ; EC = effective concentration; FHM = fathead m i nnow; FT = flow-through m e thod; FW = fresh water; H = hardness in m g CaCO 3 /L; H? = hardness unknown; LOEC = lowest-observed-effect concentration; M = m easured concentrations; n# = sam p le size used in calculation of the m ean; nm = not m easured; NOEC = no-observed-effect concentration; OW = Office of Water; P = predicted; RT = rainbow trout; S = static m e thod; SAR = structure activity relationship; SR24 = static renewal m e thod with renewals every 24 h; TOC = total organic carbon; wb = whole body; W Q C = water quality criterion; August, 1997

22Category: Aminobenz o thiaz o le Az o Dyes Hum an Health Environm ental Toxicity

Definition

. A ny disperse azo dye containing the substructure 2-am inobenzothiazole or 2-(pam ino)phenylbenzothiazole, or phenyl ring-substituted derivatives of the sam e , is considered to be a m e m b er of the category.

Haz a rd Concerns

. T here are oncogenicity/m utagenicity concerns for intact am inobenzothiazole azo dyes by analogy to dim e thyl am inostyryl benzothiazole and Butter Yellow-type dyes, such as 4-ethyl-N,N-diethylam inoazobenzene. In addition, there are liver and thyroid concerns for reduction products by an alogy to 2-am inothiazole, and neuro-toxicity concerns by analogy to chlorinated 2-am inobenzothiazole. Ecotoxicity concerns are generally chronic concerns only and are based on QSAR predictions for neutral organic com pounds. Neutral organic com pounds are an established ecotoxicity category, of which these dyes are part of the disperse dye subclass. Boundaries

. T he boundaries are not strictly defined. For a typical m e m b er of the category, R = N- and/or ring substituted p-am inophenyl groups, and R2 1 = halogens or nitro groups. General Testing Strategy

The New Chem icals Program considers the following tests to be the m o st appropriate for am inobenzothiazole azo dyes found to pose an unreasonable risk:

- Invivo m ouse m i cronucleus assay in bone m arrow by the i.p. route (40 CFR 798.5395). - 90-day subchronic toxicity test in rats by the oral route, with special attention to the thyroid and liver (40 CFR 798.2650).

23- Fish early life stage test (40 CFR 797.1600). Chronic daphnid test (40 CFR 797.1350). Algae toxicity test (40 CFR 797.1050). - If the m ouse m i cronucleus assay is positive, further characterization of the potential cancer risk m ay be recomm e nded: 2-year cancer bioassay by the oral route in 2 species of rodents (40 CFR 798.3260). Potentially significant ecotoxicity risk, as well as hum an health risks, resulting from releases to water ONLY , m ay be addressed by environm ental fate testing prior to the chronic ecotoxicity and hum an health testing. The results of the environm ental fate testing m ay preclude the need for further testing. - Jar test to determ ine the settling rate and extent of rem oval of suspended solids f r om solution (draft protocol available). If

the results of the jar test do not m itigate the health and/or ecotoxicity concerns, f urther characterization of the environm ental fate of the dye m ay be recomm e nded. - Coupled units test (40 CFR 796.3300).
January, 1992.

24Category: Anhydrides, Carboxylic Acid Hum an Health Definition. Any m o lecular structure containing one or m o re carboxylic acid anhydride groups is considered to be a m e m b er of the category for new chem ical purposes. Mem b ers of the class include new carboxylic acid anhydrides as well as new oligom ers, polym ers, prepolym ers, or reaction products of existing carboxylic acid anhydrides. As illustrated below, a typical new chem ical carboxylic acid anhydride of concern is a polym er or oligom er containing a m onom er such as m a leic anhydride.

Haz a rd Concerns. Carboxylic acid anhydrides are of concern for potential pulm onary sensitization based on data for phthalic, trim ellitic, isopropylidene bis(phthalic), and sulfonyl bis(phthalic) anhydrides. Carboxylic acid anhydrides are also of concern for potential developm ental or reproductive toxicity based on data for m a leic, succinic, and phthalic anhydrides.

Boundaries. Structures with a carboxylic acid anhydride equivalent weight of > 5,000 are presum ed not to pose a hazard under any conditions. Typically, concerns for health effects are confined to those species with m o lecular weights <1,000. The new chem ical program has thus far been concerned only with those carboxylic acid anhydrides with potentially significant inhalation exposure, and has pursued developm ental toxicity only for low m o lecular weight (<500) carboxylic acid anhydrides which are thought to have a significant potential for system ic uptake via the lung. For pulm onary sensitization, extensive system ic uptake m ay not be necessary for a biological response and we have therefore sought to regulate som e oligom eric or polym eric carboxylic acid anhydrides only for that effect. General Testing Strategy The following tests are usually prescribed for carboxylic acid anhydrides found to pose a potentially unreasonable risk:

- P ulm onary sensitization by either the m e thod of Karol (Toxicol. Appl. Pharm a col. 68:229-241, 1983), or an equivalent m e thod.

- Oral developm ental toxicity study in two species (40 CFR §798.4900). May, 1991

25Category: Anilines Environm ental Toxicity This category includes all anilines, both m onoanilines and polyanilines. It is assum e d that these com pounds need to be absorbed to be toxic, therefore, com pounds with MW s > 1000 will be excluded f r om this category. Acute toxicity f o r anilines which are liquids at room tem p erature is known to be lim ited by the octanol/water partition coefficient (K ow ). Above a log K owvalue of => 7.38, anilines show no effects at saturation during 96-h exposures (Veith and Broderius (1987). Anilines which are solids at room tem p erature m ay show no toxicity at saturation at lower Kow values depending on the m e lting point, i.e., the higher the m e lting point at a given K ow , the greater the likelihood that no toxicity will be observed at saturation. For solids, the no effects at saturation has to be determ ined on a case-by-case basis. There are no known Kow lim its for chronic toxicity at this tim e, but it m ay not be m u ch above a log K ow= 8 for liquid anilines. Future testing will determ ine this K owlim it.
Haz a rd Concerns. The acute toxicity for anilines has been determ ined through SAR Analysis: fish 96-h LC50 (Veith and Broderius 1987); fish 14-d LC50 (Deneer et al 1987); fish 14-d LC50 (Herm e ns et al 1984); daphnids 48-h LC100 (Nendza and Seydel 1988a and 1988b); and green algal 96-h EC50 (Nendza and Seydel 1988a and 1988b).; The neutral organic SARs can be used to predict green algal 96-h EC50. Arom atic diam ines (i.e., two am ines on one benzene) and dinitroanilines are known to be m o re toxic than predicted by these SARs.
Mem b ers of this category exhibit toxicity rangi ng from low toxicity (i.e., > 100 m g /L) to high

ow , MW and substitutions (e.g., dinitroanilines). toxicity (i.e., < 1 m g /L) depending on their K Environmental Fate

. Som e recent studies have shown that certain anilines are subject to rapid direct and indirect photolysis under envir onm entally realistic conditions (Leifer 1990). Boundaries. There are no known lower boundaries. The upper boundaries will be based on Kow and MW

. Acute toxicity expected with log K ow< 7.38; no effects at saturation during 96-h exposures when log K ow>= 7.38. Chronic toxicity has no known upper bound for log K ow , but it is probably near 8. MW will be < 1000. The environm ental base set of tests will be requested f o r aquatic releases and the terrestrial base set of tests will be recomm e nded f o r terrestrial exposures. Wh en the log K owis >= 7.38, chronic toxicity testing with fish and daphnids will be recomm e nded.

26General Testing Strategy . I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity tests for fish (40 CFR 797.1400) and daphnids (40 CFR 797.1300) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should equal the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. The algal toxicity testing (40 CFR 797.1050), should be done with static m e thods; m easured concentrations.; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; the highest treatm e nt concentration on a nom inal-basis equal to the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. If there is no significant risk from the PMN after the results of the environm ental base set have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Direct and Indirect Photolysis Screening Test (40 CFR 796.3765). If t1⁄2 < 2 days, go to Tier 3; if t1⁄2 > 2 days, go to Tier 4. Tier 3a. If t1⁄2 < 2 days and photolysis products are known and/or identified, then assess photolysis products for environm ental hazards. Tier 3b. If t1⁄2 < 2 days and photolysis products are not known and/or identifiable, then prepare a stock solution of PMN using the standard hum ic-containing solution described in the direct and indirect photolysis screening te st [40.796.3765 (b)(2) and (c)(2)], expose to sunlight for at least 6 half-lives (t1⁄2), and test photolysis products for toxicity with m o st sensitive species f r om environm ental base set. For exam ple, the m o st sensitive species f r om the environm ental base set has an EC50 value = 2.0 m g PMN/L (based on 100% active ingredients [AI]), therefore, prepare a 5.0 m g PMN per liter stock solution based on 100% AI using the standard hum iccontaining solution. This stock solution is exposed to sunlight for at least 6 half-lives to ensure that all of the PMN has been photolyzed, and then this stock solution is used to retest the m o st sensitive aquatic species to determ ine if the photolysis products of the PMN are m o re or less toxic that the PMN.

27Tier 4. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (40 CFR 797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test because Van Leeuwen et al ( 1990) have dem onstrated that the 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen); Daphnid chronic toxicity testing ( 40 CFR 797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; m easured TOC of dilution water in the control; the highest treatm ent concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test because Van Leeuwen et al (1990) have dem onstrated that the fish 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the fish 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen). Aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Aerobic Aquatic Biodegradation 40 CFR 796.3100 Modified Sturm Test 40 CFR 796.3260 Closed Bottle Test 40 CFR 796.3200 Modified OECD Screening Test 40 CFR 796.3240 Modified MITI Test (I) 40 CFR 796.3220 Modified AFNOR Test 40 CFR 796.3180
II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial exposures. Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay.

28References

. Deneer JW , Sinnige TL, Seinen W and Herm ens JLM. 1987. Quantitative structure-activity relationships for the toxicity and bioconcentration factor of nitrobenzene derivatives towards the guppy ( Poeciliareticulata ). Aquatic Toxicology 10:115-129.
Herm ens J, Leeuwangh P, and Musch A. 1984. Quantitative structure-activity relationships and m i xture toxicity studies of chloro- and alkylanilines at an acute lethal toxicity level to the guppy, Poeciliareticulata

. Ecotoxicology and Environm ental Safety 8:388-394. Leifer A. 1990 (14 Dec). Review of Section 5 test data for o -phenylenediam i ne, m phenyldiam ine, and p -phenylenediam i ne. Mem o randum

. W a shington, DC: Exposure Assessm ent Branch, Exposure Evaluation Division (TS-798), Office of Toxic Substances, United Stated Environm ental Protection Agency, 401 M St, SW , 20460-0001. Nendza M and Seydel JK. 1988a. Multivariate data analysis of various biological test system s used for the quantification of ecotoxic com pounds. Quantitative Structure-Activity Relationships 7:165-174. Nendza M and Seydel JK. 1988b. Quantitative structure-activity relationships for ecotoxicologically relevant biotest system s and chem icals. Chem osphere 17:1585-1602. Van Leeuwen CJ, Adem a DMM, and Herm ens J. 1990. Quantitative structure-activity relationships for fish early life stag e toxicity. Aquatic Toxicology 16:321-334. Veith GD and Broderius SJ. 1987. Structure-toxi city relationships for industrial chem icals causing type (II) narcosis syndrom e. In Kaiser KLE (ed), QSAR In Environm ental Toxicology - II, p 385-391. Reidel Publishing Com p any. May, 1991

29Subcategory of Anilines: Dianilines Hum an Health Environm ental Toxicity Definition. A PMN m u st have the following m i nim um structural requirem e nts to be considered a m e m b er of this category:

-it m u st have at least two phenyl rings w ith a bridging carbon, oxygen, nitrogen, or sulfur -each term inal phenyl ring m u st have a prim ary am ino group (or a group that can be readily m e tabolized to a prim ary am ino group) either m e ta- or para- to the bridging atom Com pounds with one or m o re additional phenyl ring(s), with or without ring substituents, and one or m o re bridging atom s are also included in the category.

Minim um Structure f o r Dianiline Category

The com pounds of greatest concern are those having X = C, N, or O and n = 0 or 1.
Haz a rd Concerns: Mem b ers of the class are considered to be potential carcinogens and m

utagens by analogy to 4,4' -m ethylendianiline, 4,4' -m ethylene bis(o-toluidine), and 4,4'

- oxydianiline. Class m e m b ers are potential retinotoxic agents by analogy to 4,4'

- m e thylenedianiline, 4,4' -oxydianiline, and the diam inodiphenyl alkane drugs and are also potential reproductive and system ic toxicants by analogy to 4,4' -m ethylenedianiline.

30General Testing Strategy The New Chem icals Program has considered the following toxicity tests to be the m o st appropriate for dianilines found to pose an unreasonable risk:
I. Exposure to Hum ans - rat acute oral retinopathy screening study (protocol to be approved by EPA) - pigm ented rat 90-day subchronic toxicity study by the oral (40 CFR 798.2650) or inhalation (40 CFR 798.2450) route to include histopathological exam ination of the eyes and reproductive organs (eyes to be exam ined by both light and electron m i croscopy) - 2-year carcinogenicity bioassay (40 CFR 798.3300) in rats and m i ce. In the 90-day subchronic toxicity study, the com p any m ay opt to carry a group of anim als at each dose level for a 90-day recovery period to determ ine whether potential retinopathy is reversible.
For som e com pounds in this class, short-term m utagenicity testing m ay be appropriate. Wh en appropriate, specif i c testing will be determ ined by EPA m utagenicity assessors on a case by case basis by referral to "short question."
Wh ere the general population is at a significant risk from drinking water exposure (via surface water releases) to PMN chem icals of this class, the following fate testing m ay be appropriate: -direct and indirect photolysis 40 CFR 796.3765 and 796.3700 -aerobic aquatic biodegradation 40 CFR 796.3100 -see aniline category description f o r details Results of the above fate tests m ay m itigate concern for drinking water exposure to PMN of this class. However, fate test results W I LL NOT CHANGE AGENCY CONCERNS FOR OCCUPATIONAL EXPOSURE to PMNs of this category.
II. Release to Aquatic Ecosystem s Tier 1. Because of the above boundary conditions, and the need to assess hum an exposure as well as environm ental toxicity, if there is insufficient knowledge about the water solubility of the dye, then it should be m easured (40 CFR §796.1860). The fish and daphnid acute toxicity tests f r om the aquatic base set of environm ental toxicity tests will be recomm e nded for aquatic exposures. The acute toxicity tests for fish (40 CFR §797.1400) and

31daphnids (40 CFR §797.1300) will be done using the flow-through m e thod with m easured concentrations, and effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations. If there is no significant risk from the PMN after the results of the fish and daphnid acute toxicity tests have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Direct and Indirect Photolysis Screening Test (40 CFR 796.3765). If t1⁄2 < 2 days, go to Tier 3; if t1⁄2 > 2 days, go to Tier 4. Tier 3a. If t1⁄2 < 2 days and photolysis products are known and/or identified, then assess photolysis products for environm ental hazards. Tier 3b. If t1⁄2 < 2 days and photolysis products are not known and/or identifiable, then prepare a stock solution of PMN using the standard hum ic-containing solution described in the direct and indirect photolysis screening test [40 CFR 796.3765 (b)(2) and (c)(2)], expose to sunlight for at least 6 half-lives (t1⁄2), and test photolysis products for toxicity with m o st sensitive species from environm ental base set. Tier 4. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (40 CFR 797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; and statistical analysis of effective concentrations at days 7, 14, 21, and 28. Daphnid chronic toxicity testing ( 40 CFR 797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21. Aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Aerobic Aquatic Biodegradation Modified Sturm Test Closed Bottle Test Modified OECD Screening Test Modified MITI Test (I) Modified AFNOR Test
III. Release to terrestrial Ecosystem s : 40 CFR 796.3100 40 CFR 796.3260 40 CFR 796.3200 40 CFR 796.3240 40 CFR 796.3220 40 CFR 796.3180

The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded f o r terrestrial exposures. Chronic toxicity testing fo r terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay.

32References Am erican Dye Manufacturers Institute, Inc. (1974). "Dyes and the Environm ent: Reports on Selected Dyes and Their Effects," Vol. II, ADMI, New York. Auer, C.M., Nabholz, J.V. and Baetcke, K.P. (1990). "Mode of Action and the Assessm ent of Chem ical Hazards in the Presence of Lim ited Data: Use of Structure-Activity Relationships (SAR) under TSCA, Section 5," Environm ental Health Perspectives , Vol. 87, pp. 183-197. Little, L.W ., and Lam b J.C., III (1972). "Acute Toxicity of 46 Selected Dyes to the Fathead Minnow, Pim e phales prom elas , "Final Report to the Am erican Dye Manuf acturers Institute, Inc., UNC W a stewater Research Center, Departm e nt of Environm ental Sciences and Engineering, School of Public Health, University of North Carolina, Chapel Hill, NC. Nabholz, J.V. (1990). "The OTS PMN ECOTOX Data Base: a Confidential Business Inform ation (CBI) Collection of Environm ental Toxicity Data from New Chem ical Subm issions Under Sec. 5," Unpublished, Environm ental Eff ects Branch, Health and Environm ental Review Division (TS-796), U. S. Environm enta l Protection Agency, W a shington, DC. Sigm an, C.C., Helm s, C.T., Papa, P.A., Atkinson, D.L., Doeltz, M.K., and W i nship-Ball, A. (Jan. 1983). "Anthraquinone Dyes and Related Chem ical s: Review and Assessm ent of Potential Environm ental and Health Aspects," Final Report to the Dyes Environm ental and Toxicology Organization, Inc., SRI International, Menlo Park, CA. Tonogai, Y., Ito, Y., Iwaida, M., Tati, M., Ose, Y., and Sato, T. (1979). "Studies on the Toxicity of Coal-Tar Dyes. II. Exam ination of the Biol ogical Reaction of Coal-Tar Dyes to Vital Body," The Journal of Toxicological Sciences , Vol. 4, pp. 211-219.
August 1991; revised January 1995; revised October 1995

33Category: Anionic Surfactants Environm ental Toxicity Definition. Any m o lecular structure with a net negative charge and having surfactant activity is a m e m b er of this category. The category includes for exam ple, alkyl sulfonates, alkyl benzene sulfonates, alkyl silicic acids, alkyl phosphates, alkyl carboxylic acids, or com b inations of these anionic groups, e.g., alkyl sulfonate with carboxylic acid substitutions.

Haz a rd Concerns. Anionic surfactants are toxic to a wide variety of aquatic organism s. Toxicity increases exponentially with increasing carbon chain length up to 16 carbons [for linear alkyl sulfonate (LAS) surfactants], and then decr eases with additional carbons. At a chain length of 16 carbons, fish and daphnids are m o st sensitive, but at lower chain lengths green algae are m o re sensitive than fish and daphnids. There are a num ber of structure-activity relationships (SARs) to predict the toxicity of m e m b ers of this category. Most are parabolic regression equations and others rely on nearest analog m e thodology. Boundaries. There is no m o lecular weight boundary. Surfactants with m o lecular weights > 1000 are still toxic. Acute toxicity m ay be low, i.e., > 100 m g /L, if a strong cationic surfactant counterion and the anionic surfactant form a tight ion pair. Testing. To address ecotoxicity concerns, base set acute aquatic toxicity testing (green algae: static m e thod, daphnid and fish: flow -through m e thod, all m easured concentrations). Tier 1. The acute aquatic base setof environm ental toxicity tests will be recomm e nded for aquatic exposures and the terrestrial base setof environm ental toxicity tests (i.e., the early seeding growth test, the earthworm acute toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded for any terrestrial exposures. Acute fish toxicity test Acute daphnid toxicity test Green algae toxicity test Early seedling growth test Earthworm acute toxicity test Soil m i crobial comm unity bioassay OPPTS 850.1075 OPPTS 850.1010 OPPTS 850.5400 OPPTS 850.4230 OPPTS 850.6200 O PPTS 850.5100

34Tier 2. If acute toxicity testing indicates a significant risk, then environm ental fate testing in the form of aerobic biodegradation testing is recomm e nded. Aerobic biodegradability can be determ ined using one of the following test guidelines, listed in approxim a te order of preference: CO 2 evolution Closed bottle Modified OECD screening Modified MITI (I) DOC die-away Manom etric respirom etry O PPTS 835.3110 OPPTS 835.3110 O PPTS 835.3110 OPPTS 835.3110 OPPTS 835.3110 OPPTS 835.3110 Tier 3. In addition, if acute toxicity testi ng indicates a significant risk, then chronic aquatic toxicity testing with f i sh and aquatic invertebrates will be recomm e nded. Fish early life stage test OPPTS 850.1400 Daphnid chronic toxicity testing O PPTS 850.1300
September 1988, revised September, 1996

35Category: Az ides Environm ental Toxicity Azides are a class of chem icals consisting of both inorganic and organic com pounds and characterized by the functional group N=N=N. Substitutions m ay be either m e tals or organic com pounds, e.g., acetic acid and benzene. So far, only m onom eric arom atic azide com pounds have been subm itted as PMNs. No aliphatic azides, e.g., alkyl azides, have been subm itted as PMNs. Aliphatic azides m ay be too explosive to isolate, however, diazidoacetic acid ester is known to exist and has been tested in rats.
Haz a rd Concerns: It is assum e d that azides have to be absorbed to be toxic, therefore, azides with MW >1000 will be excluded from this category. Acute toxicity for m onom eric azides is assum e d to be correlated and lim ited by the octanol/water partition coef f i cient (K ow ). Above a log K owvalue of >5.0, azides are assum e d to show no effects at saturation during 96-hour exposures to fish. Organic azides which are solid s at room tem p erature m ay show no toxicity at saturation at lower K owvalues depending on their m e lting point and/or their water solubility. For exam ple, for organic azides, the higher the m e lting point at a given Kow , the greater the likelihood that no toxicity will be occur at saturation. For solids, no effects at saturation has to be determ ined on a case-by-case basis. There are no m easured upper K owlim its f o r chronic toxicity at this tim e, but it m ay not be m u ch above a log K ow= 8. Future testing will determ ine K owlim its. The aquatic toxicity of organic azides has been determ ined through SAR Analysis: log fish 96-h LC 50 (m illim oles/L) = -1.881 -0.124 log K ow where n=2, R2 =1.0, LOGKOW (SRC)<5, MW <1000; The toxicity for inorganic azides has to be determ ined by integrating the toxicity of Na azide with the toxicity of any other inorganic elem ent, e.g., Pb, with a MW adjustm e nt for differences in toxicity between Na azide and th e other inorganic elem ent. Na azide [26628-22-8] has been tested by the USEPA ERL-Duluth: fathead m i nnow 96-h LC 50 = 5.46 m g /L based on a flow-through m e thod with 18 volum e changes per day and nom inal concentrations. The SAR for organic azides is based on (1) this toxicity value, (2) a calculated log K owvalue of -6.3 using SRC's LOGKOW , and (3) no acute effects to fish at saturation assum e d to occur at a log Kow > 5.0 which is equivalent to a log fish 96-h LC 50 = -2.5 in m illim oles per liter. CLOGP has a m i ssing fragm e nt constant for the azide group. Environmental Fate: Na azide is known to hydrolyze in water to form hydrazoic acid (or , [7782-79-8], MW 43, M12,4815). Hydrogen azide is extrem ely explosive according to Merck. In addition, azides can be transform e d to nitrenes by uvlight. hydrogen azide, NH 3

Boundaries: MW <1000. Log K ow 50 ; and log K owassum e d to be <8.0 for chronic toxicity to aquatic organism s, but could be higher. <5.0 for acute toxicity to fish and aquatic invertebrates; log K ow<6.4 for toxicity to green algae as a 96-h EC

36General Testing Strategy : I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity test s for fish (OPPTS 850.1075) and daphnids (OPPTS 850.1010) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (ai) and m ean m easured concentrations; m easured TOC of dilution water in the control; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; and solvent can be used to assist the aldehyde to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the test chem ical significantly above its aqueous solubility lim it. The algal toxicity testing (OPPTS 850.5400), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (ai) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; and solvent can be used to assist the aldehyde to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the test chem ical significantly above its aqueous solubility lim it. If there is no significant risk from the PMN chem ical after the results of the environm ental base set have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Aerobic biodegradabilityaccording to either of the following test guidelines: Ready Biodegradability OPPTS 835.3110 Sealed Vessel CO2 Production Test OPPTS 835.3120 If there is no significant risk from the PMN chem ical after the results of biodegradation testing have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 3. Tier 3. Fish chronic toxicity testing , i.e., f i sh early lif e stage (ELS) toxicity testing (OPPTS 850.1400), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrati ons at days 7, 14, 21, and 28; m e asured TOC of dilution water in the control; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN significantly above its aqueous solubility lim it; and the 7-d ELS

37stage toxicity test cannot be substituted for the 28-d ELS toxicity test because the 7-d ELS toxicity test m ay underestim ate chronic toxicity m easured by the 28-d ELS toxicity test when the Chronic Values are com p ared. Daphnid chronic toxicity testing (O PPTS 850.1300), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; m easured TOC of dilution water in the control; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN significantly above its aqueous solubility lim it; and the 7-d daphnid short-term chronic toxicity test cannot be substituted for the 21-d toxicity test because the daphnid 7-d short-term toxicity test m ay underestim ate chronic toxicity m easured by the 21-d toxicity test when the chronic values are com p ared.
II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test [O PPTS 850.4230], the earthworm toxicity test [OPPTS 850.6200], and the soil m i crobial comm unity bioassay [OPPTS 850.5100]) will be recomm e nded for terrestrial exposures. Chronic toxicity te sting for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay. References: L’Abbe, G. (1969). Decom position and addition r eactions of organic azides. Chem ical Review, 69, 345-363. Scriven, E.F. (Ed.). (1984). Azides and nitrenes: Reactivity and utility. pp.95-204. New York: Academ ic Press.
Septem ber 1996

38Category: Benz otriaz oles Environm ental Toxicity Definition. This category includes all benzotriazoles.
Haz a rd Concerns. The toxicity for benzotriazoles with -NH has been determ ined through SAR Analysis (Clem e nts 1988). Benzotriazoles with -NH are known to be m o re toxic than neutral organic chem icals, and this excess toxicity decreases with increasing K ow . The toxicity of benzotriazoles with aliphatic substitutions on the nitrogen are expected to act like neutral organic com pounds. Other substitutions on the N, e.g., -SH, have to be evaluated on a case-by-case basis.
Mem b ers of this category exhibit toxicity rangi ng from low toxicity (i.e., > 100 m g /L) to high

ow , MW , and m e lting point. toxicity (i.e., < 1 m g /L) depending on their K Boundaries

. There are no known lower boundaries. The upper boundaries will be based on K ow , MW , and m e lting point (m p). These com pounds need to be absorbed to be toxic, therefore, com pounds with MW s < 1000 are expected to cause toxicity, while com pounds with MW s > 1000 are no expected to show toxicity at saturation. Acute toxicity for benzotriazoles which are liquids at room tem p erature is known to be lim ited by the octanol/water partition coef f i cient (Kow ). Above a log K owvalue of 5.0, benzotriazoles show no effects at saturation during 96-h exposur es (Clem e nts 1988). Benzotriazoles which are solids at room tem p erature m ay show no toxicity at saturation at lower K owvalues depending on the m e lting point, i.e., the higher the m e lting point at a given K ow , the greater the likelihood that no acute toxicity will be observed at saturation. For solids, the no-effects-at-saturation point for acute toxicity has to be determ ined on a case-by-case basis. The K owlim it f o r chronic toxicity is set at a log K ow chronic toxicity testing will determ ine the Kowlim it. = 8 for liquid benzotriazoles. For solids,
Therefore, acute toxicity is expected when log K ow=< 5.0 and MW < 1000; no effects at saturation during 96-h exposures are expected when log K ow> 5.0 and MW < 1000. The upper boundary for chronic toxicity is 8.0. Only chronic toxicity is expected when log K ow> 5.0 and < 8.0, and MW < 1000. Wh enever MW > 1000, no effects are expected at saturation because it is assum e d that benzotriazoles have to be absorbed to be toxic. Aerobic biodegradation is expected to be the dom inant route of transform a tion in the environm ent.

39General Testing Strategy . I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity tests for fish (40 CFR §797.1400) and daphnids (40 CFR §797.1300) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested chem ical; and solvent can be used to assist the chem ical to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the chem ical beyond its aqueous solubility lim it. The algal toxicity test (40 CFR §797.1050) should be done with the static m e thod; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the chem ical; and solvent can be used to assist the chem ical to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the chem ical beyond its aqueous solubility lim it. If there is no significant risk from the benzotriazole after the results of the environm ental base set have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Aerobic Aquatic Biodegradation 40 CFR 796.3100 Modified Sturm Test 40 CFR 796.3260 Closed Bottle Test 40 CFR 796.3200 Modified OECD Screening Test 40 CFR 796.3240 Modified MITI Test (I) 40 CFR 796.3220 Modified AFNOR Test 40 CFR 796.3180 If there is no significant risk from the benzotriazole after the results of the aerobic biodegradation testing have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 3. Tier 3. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (40 CFR §797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it for the chem ical; solvent can be used to assist the chem ical to reach its aqueous solubility lim it quicker, but cannot
40be used to artificially enhance the water solubility of the chem ical beyond its aqueous solubility lim it; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test because Van Leeuwen et al (1990) have dem onstrated that the 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen). Both anilines and benzotriazoles with -NH are m o re toxic than predicted based on narcosis alone, i.e., both benzotriazoles and anilines have excess toxicity due to a m o re specific m ode(s) of toxic action. A seven day exposure m ay not allow enough tim e for this excess toxicity to be expressed either because of not enough exposure and/or not enough tim e for m e tabolic activation. Daphnid chronic toxicity testing ( 40 CFR §797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the chem ical; solvent can be used to assist the chem ical to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the chem ical above its aqueous solubility lim it; and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test (Van Leeuwen et al 1990).
II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test, the avian acute oral toxicity test, and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial exposures. Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, the avian reproductive toxicity test, and the soil m i crobial comm unity bioassay. In addition, the fate test for Inherent Biodegradability in soil (40 CFR §796.3400) m ay be recomm e nded. References Clem ents, RG (editor). 1988. Estim ating toxicity of industrial chem icals to aquatic organism s using structure-activity relationships. EP A-560-6-88-001. W a shington, DC: Environm ental Effects Branch, Health and Environm ental Revi ew Division, Office of Toxic Substances (TS796), United States Environm ental Protection Ag ency. Available from the National Technical Inform ation Service, Springfield, VA 22161, PB89-117592. Van Leeuwen CJ, Adem a DMM, and Herm ens J. 1990. Quantitative structure-activity relationships for fish early life stag e toxicity. Aquatic Toxicology 16:321-334. March, 1992

41Category: Benz otriaz ole-hindered phenols Hum an Health Environm ental Toxicity (see “Phenols”) The predom inant use of m e m b ers of this category is as a UV light stabilizer in coating or plastic form ulations. Definition. Any m o lecular structure containing the following substructure is considered to be a m e m b er of the category.

Note : Health ef f ects m ay vary depending on the nature of the ring substituents.
Haz a rd Concerns. I. Health System ic toxicitybased on 8(e) and PMN data on analogues. These data are generally consistent across com pounds, with the following endpoints occurring in subacute, subchronic, and chronic oral feeding studies in rats and dogs in which doses ranged from 5 to 15,000 ppm

. Increased organ weights (liver and kidney, w ith associated histopathology at higher doses); hem a tological effects (decreased hem oglobin, packed cell volum e, and erythrocytes); and imm une system effects (weight changes in thym us, spleen, lym ph nodes; decreased leukocytes). Generally, treatm e nt-related effects are dose-related, increasing in frequency and severity with increasing dose. Males appear to be m o re sensitive than f e m a les, and sim ilar toxic ef f ects occur in both rats and dogs. The lowest available NOAEL for system ic effects, from a 90-day rat study, is 5 ppm , or 0.25 mg/kg/day

. Reproductive toxicity- Concern is also based on 8(e) and PMN data on analogues. As with

42system ic toxiciy, the data are generally consistent, with the following endpoints occurring in subacute and subchronic oral feeding studies in rats and dogs in which doses ranged from 5 to 15,000 ppm

. Atrophy of the sem i nal vesicles, significant reduction in absolute and relative testes weight, significant reduction in absolute and relative prostate weight, and abnorm a l sperm a togenesis. A subchronic dog study also provided evidence of fem a le reproductive toxicity. Generally, treatm e nt-related effects are dose-related, increasing in both frequency and severity with increasing dose. Males appear to be m o re sensitive than f e m a les, and sim ilar toxic ef f ects occur in both rats and dogs. The lowest NOAEL for reproductive toxicity, from a 90-day dog study, is 15 mg/kg/day

. Derm al sensitization- Derm al sensitization test data were positive in 3/4 com pounds tested. Two of the com pounds showed strong sensitizing potential based on results obtained from m a xim i zation tests in which 70% and 90% of the treated guinea pigs becam e sensitized, respectively.
II. Environm ental - The environm ental concerns for the substances described in this category have already been described in the Phenolcategory. Boundaries. Not enough is known about the structure-activity relationships of these com pounds to set any boundaries on m e m b ers of the category; com pounds with both "sm a ll" and "large" substituents have shown effects, and the effects are generally consistent from com pound to com pound. General Testing Strategy I. Health System ic Toxicity and Reproductive Toxicity

. A 90-day gavage study in rats with special attention to hem a tology; weight and histopathology of lym phoid organs (spleen, thym us, and bone m arrow); cellularity of the bone m arrow, thym us, and spleen; and histopathology of the liver, kidney, heart, and all endocrine glands fo r which weight changes are observed. Particular attention should be directed toward achieving satisfactory quality from fixation and em bedding of

the testes, f o llowing the recomm e ndations of Russell et al. [ R ussell LD, Ettlin RA, Sinha Hikim AP, Clegg ED. 1990. Histological and hist opathological evaluation of the testis. Cache River Press, Clearwater, FL]. A gavage study, as opposed to an oral feeding study, is recomm e nded because decreased body weight gain, final body weight, and/or food c onsum ption have been reported in som e oral feeding studies with analogues.

43If imm unopathology and hem a tology in the 90-day study support the concern for adverse effects of the PMN substances on the imm une system , then a further battery of tests, such as the NTP' s Imm unotoxicology Tier II Screening Panel, will be recomm e nded.
Derm al Sensitization

. Because derm al sensitization is not routinely pursued under TSCA 5(e) as a regulatory endpoint, testing is not recomm e nded. A letter be sent to the PMN subm itter expressing the Agency' s belief that the com pounds are likely to be derm al sensitizers and that protective m easures should be taken for workers.
II. Environm ental toxicity If exposure to the aquatic environm ent dem onstrates a potential risk, testing of the substances described in this category is the sam e as that prescribed for the Phenols category. June, 1993.

44Category: Boron Compounds Human Health Environmental Toxicity Definition. This category includes borates, organoborates, borate esters, boron hydrides, boranes, and boroxines.
Hum an Health : Haz a rd Concerns. Reproductive toxicity (m ales/fem ales), blood toxicity, and neurotoxicity

. The Agency has an oral RfD for boron of 0.09 mg/kg/day (uncertainty factor 100) based on the finding of testicular atrophy and sperm a togenic arrest in dogs in a 2-yr feeding study (W eir and Fisher 1972, as summ arized in IRIS 1995). [All toxicity inform ation presented here are summ arized in an assessm ent of various aqueous cleaner chem icals prepared in 1990 for the Office of Air and Radiation. Reproductive toxic ity inform ation also summ arized in IRIS 1995.] Reproductive Toxicity

. The m o st significant effect seen in anim als is reproductive toxicity (i.e., sterility in m a les and fem a les, and testicular atrophy in m a les). A three-generation reproductive study in rats dem onstrated sterility in m a les (lack of sperm a tozoa in atrophied testes) and fem a les (decreased ovulation) after exposure to 58.5 m g boron/kg in the diet. NOAEL (rats) 17.5 m g boron/kg

. Sterility was also found in m a les (at 50 or 100 m g boron/kg) in a serial m a ting adjunct to a 60-day feeding study; NOAEL (rats) 25 m g boron/kg

. Data from oral subchronic and chronic studies in rats, dogs, and m i ce also provide evidence that boron produces adverse effects on the m a le reproductive system

. A statistically significant reduction in relative and absolute testes weights was seen in a 2-yr and a 90-day dietary study in m a le rats at 58.5 and 26.3 m g boron/kg, respectively. NOAEL’s (rats) 17.5 and 8.8 m g boron/kg, respectively

. In m a le rats, dietary exposure to 50 m g /kg boron resulted in a significant reduction in f e rtility, a signif i cant reduction in epididym al weight, and a reduction in sperm num ber after 30 days of treatm e nt. A significan t reduction in testicular weight and further reduction in sperm occurred after 60 days of treatm e nt. NOAEL (rats) 25 m g boron/kg

. Sim ilar results were observed in several drinking water studies in rats.
In m a le dogs, dietary exposure to 29.4 m g boron/kg for 2 yr resulted in severe testicular atrophy and sperm a togenic arrest. NOAEL (dogs) 8.8 m g boron/kg

. In m a le and fem a le dogs, dietary exposure to 43.8 and 4.4 m g boron/kg for 90 days did not affect the fem a le reproductive system , but produced decreased testes weights (statis tically significant at 43.8 m g boron/kg). NOAEL (dogs) 0.4 m g boron/kg

. In m a le m i ce, dietary exposure to 131 m g boron/kg for 103 wk (only dose tested) caused an increased incidence of testicular atrophy and interstitial hyperplasia. Blood Toxicity

. In a chronic feeding study in rats, packed cell volum e and hem oglobin levels were significantly decreased in m a les and fem a les at 58.5 m g boron/kg. NOAEL ( rats) 17.5 m g boron/kg

. In a subchronic drinking water study in rats, a significant decrease in plasm a triglyceride levels was noted at 23.7 and 47.4 m g boron/kg (only doses tested). LOAEL (rats) 23.7 m g boron/kg

. In a subchronic feeding study in dogs, 43.8 m g boron/kg but not 4.4 or 0.4

45m g boron/kg produced decreased packed cell volum es and hem oglobin levels. NOAEL (dogs) 4.4 m g boron/kg

. In a subchronic feeding study in m i ce, m i nim a l to m ild extram edullary hem a topoiesis of the spleen was observed in all dosed groups (ca. 34, 68, 136, 272, or 544 m g boron/kg). LOAEL (m ice) 34 m g boron/kg

. There is also a case study in which an infant exposed to 43 m g boron/kg/day for 12 weeks developed anem ia. Neurotoxicity

. Ingestion of borax by hum an infants results in neurotoxicity. Case reports of nine infants who were exposed by sucking pacifiers dipped in a borax-honey m i xture docum ent sim ilar findings of neurological sym p tom s (not sp ecified) and seizures. For seven infants, doses are reported as 2 to 3 g borax/wk for 4 to 10 wk (ca. 4 to 16 m g boron/kg/day ). Serum boron levels m easured in three of the seven infants were elevated. For the other two infants, doses are reported as 125 m g borax/kg over a 12-wk period and 9 g borax over a 5-wk period (ca. 43 and 8 m g boron/kg/day , respectively). The effects appear to be reversible upon cessation of exposure. Neurotoxicity testing is not recomm e nded because anim al studies do not appear to be predictive of this effect. Boundaries. The boundaries of this category are not yet well defined for health effects. All of the toxicity data are on boric acid or borax. A m o lecular weight cutoff of 1,000 is proposed, but there is no inf o rm ation f r om available toxicity data on a proper water solubility cutof f

. General Testing Strategy. The OECD reproductive toxicity screen (OECD 421) with special additional attention to hem a tology. If this screen is positive for reproductive toxicity, a reproductive fertility study in rats according to the OPPTS Harm onized Test Guidelines (870.3800) is recomm e nded. In cases in which exposures are exclusively derm al, a derm al absorption study (either in vivo or in vitro) could be conducted to refine the derm al risk assessm ent. If the in vivo study is selected, the test m a terial should not be corrosive. If there is som e uncertainty as to the corrosivity of the neat test m a terial, a derm al irritation study should be undertaken first. If the neat m a terial is corrosive, dilution to a noncorrosive concentration with a vehicle such as acetone or the alcohol used to m ake the ester is recomm e nded. Environm ental Toxicity : Most of the toxicity inform ation for boron com pounds is for boric acid and sodium tetraborate. These boron com pounds show low acute toxicity towards fish (250.0 m g /L as a m ean acute toxicity value) and daphnids (70.0 m g /L as a m ean acute toxicity value), but m oderate toxicity towards green algae (20.0 m g /L as a m ean EC50 value). However, these com pounds exhibit large acute to chronic ratios (ACR) towards fish (125 as a m ean ACR) and the difference between the EC50 value and the chronic value (ChV) in green algae is large (about 117 as a m ean difference). The m ean ACR for daphnids is 8. The m ean ChVs for fish, daphnids, and green algae are 2.0 m g /L, 9.0 m g /L, and 0.170 m g /L, respectively. Green algae appears to be the m o st sensitive group of species with respect to boric acid and sodium tetraborate.

46There are only screening data for fish for borate esters, specifically, only for tributylborate. These screening data showed no effects at 10.0 m g /L for 24 hours towards three species of fish. However, the purity of the tributylborate was not given. The only data for boranes are for tert-butylam ine borane. These data are: fish 96-h LC50 = 13.0 m g /L, daphnid 96-h LC50 = 0.700 m g /L, green algal 7-d EC50 = 3.0 m g /L, and algal ChV = 0.300 m g /L. The only toxicity inform ation for boron hydrides are for m amm a ls. These data show that all boron hydrides are highly toxic and m o re toxic than borates. The m a jor environm ental hazard concerns for this category are for chronic toxicity towards fish and toxicity towards green algae. Since there is no SARs for boron com pounds, hazard profiles are developed using the nearest analog(s). Boundaries. Boron com pounds m u st have water solubilities equal to or greater than 1.0 

### : g/L (ppb) and m o lecular weights generally need to be near or below 1000. Environmental Fate. Certain borates are subject to hydrolysis under environm entally realistic conditions. General Testing Strategy. The f o llowing testing strategy will address aquatic toxicity concerns:
I. Release to Aquatic Ecosystem s : Tier 1a. If the hydrolysis rate for any m e m b er of this class is predicted to be slow (t1⁄2 > 2 days) or if hydrolysis will result in one or m o re products which are expected to be just as toxic as the parent borate, then the fish and daphnid acute toxicity tests from the aquatic base set of environm ental toxicity tests will be recomm e nded for aquatic exposures. The acute toxicity tests for fish (40 CFR 797.1400 or OPPTS 850.1075) and daphnids (40 CFR 797.1300 or OPPTS 850.1010) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should equal the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. The algal toxicity testing (40 CFR 797.1050 or OPPTS 850.5400), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective

47concentrations at 24, 48, 72, and 96 hours; test m e di um with at least 0.300 m g /L EDTA as a final concentration; the highest treatm e nt concentration on a nom inal-basis equal to the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. If there is no significant risk from the PMN after the results of the environm ental base set of tests have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 1b. If the hydrolysis rate for any m e m b er of this class is predicted to be rapid (t1⁄2 < 2 days) and if hydrolysis will result in products which are expected to be significantly less toxic as the parent borate, then hydrolysis as a function of pH and tem p erature (40 CFR 796.3510 or OPPTS 835.2130) will be recomm e nded. If the resulting hydrolysis t1⁄2 is < 2 days, then this result will be integrated into a new risk assessm ent for the PMN. However, the resulting hydrolysis t1⁄2 > 2 days, then the environm ental base set will be recomm e nded. If there is no significant risk from the PMN after the results of the environm ental base set of tests have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (40 CFR 797.1600 or OPPTS 850.1400), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; and statistical analysis of effective concentrations at days 7, 14, 21, and 28. Daphnid chronic toxicity testing ( 40 CFR 797.1330 or OPPTS 850.1300), with flowthrough m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21. Aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Aerobic Aquatic Biodegradation 40 CFR 796.3100 or OPPT 835.3100 Modified Sturm Test 40 CFR 796.3260 or OPPT 835.3110 Closed Bottle Test 40 CFR 796.3200 or OPPT 835.3110 Modified OECD Screening Test 40 CFR 796.3240 or OPPT 835.3110 Modified MITI Test (I) 40 CFR 796.3220 or OPPT 835.3110 Modified AFNOR Test 40 CFR 796.3180 or OPPT 835.3110
II. Release to terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests (i.e.,

48the early seeding growth test, the earthworm toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial expos ures. Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay. July, 1991; revised August, 1997.

49Category: Cationic Dyes Environm ental Toxicity Definition. Any dye bearing one or m o re net perm anent positive charges is considered to be a m e m b er of this class. A "typical" new chem ical of concern is a water soluble m onoazo dye with a single delocalized positive charge.
Haz a rd Concerns. W a ter soluble cationic dyes are toxic to fish, daphnids, and algae. Poorly soluble dyes tend to be toxic only to algae. No relationship between structure and activity has been found for cationic dyes. Boundaries. There are no established boundaries for cationic dyes. There is no m o lecular weight threshold - one dye with a m o lecular weight of 3,000 and bearing four positive charges was found to be toxic to fish. Testing. To adress environm ental concerns, the base set of environm ental toxicity tests (i.e., the fish acute toxicity test, the daphnid acute toxicity test and the green algal toxicity test) is recomm e nded. These tests are to be done using the static m e thod and nom inal concentrations. In addition, two additional fish acute toxicity tests are recomm e nded with known am ounts of hum ic acid added to the dilution water. This testing is necessary to m easure the m itigating effects of dissolved organic carbon (DOC) on the t oxicity of the cationic dye. One test will be done with 20 m g hum ic acid/L in dilution and the second test will have 10 m g hum ic acid/L in dilution water. Total organic carbon (TOC) will be m easured three tim es in each test and the TOC of the clean dilution water will also be m easured at the beginning of the test. If

DOC m itigates the environm ental risk to water columnorganism s, then testing using natural sedim e nts and organism s known to ingest sedim e nt m ay be recomm e nded. March, 1990

50Category: Cationic (quaternary ammonium) surfactants Environm ental Toxicity Definition. Any cationic surfactant is a m e m b er of this category, for exam ple: CH 3 | CH 3 -N + -(CH 2 -) 15 -CH 3 Br ­ | CH 3 Haz a rd Concerns. Cationic surfactants are biocidal to a wide array of species in the environm ent. Toxicity increases exponentially with increasing carbon chain length up to 16 carbons and then decreases with increasing chain length. QSAR (quantitative structure-activity relationships) have been developed to predict toxicity. Boundaries. Little toxicity is observed when the carbon chain length exceeds 22. Tight ion pairs will not be signif i cantly toxic. General Testing Strategy . To address ecotoxicity concerns, base set aquatic toxicity testing in algae, daphnids, and fish, plus hum ic acid testing in fish (20 m g /L hum ic acid in dilution water and 10 m g /L) is recomm e nded. All testing uses static m e thod, nom inal concentrations. March, 1990

51Category: Cobalt Environm ental Toxicity This category includes but is not lim ited to inorganic and organic com pounds of cobalt (Co), e.g., soluble cobalt cations, cobalt esters, and organocobalt com pounds.
Haz a rd Concerns

. Most of the toxicity inform ation for inorganic cobalt com pounds is for the soluble salts: chloride, nitrate, and sulfate. There are no known data for organoCo com pounds, organic acid chelates with Co, or for Co esters. The best toxicity data for Co are listed below with predicted (P) and m easured (M) toxicity values indicated and effective concentrations in m g Co/L (ppm ): fish (FHM) 96-h LC 50 daphnid 48-h LC 50 daphnid 48-h LC 50 W Q 2° acute value green algal 7-d EC 50 RTELS 28-d LC 50 fi sh ChEC 20 fish Chronic Value (ChV) fish ACR daphnid ChEC 20 daphnid ChV daphnid ChV daphnid ACR W Q 2° chronic value algal ChV Abbreviations: = = = = = = = = = = = = = = = 48.0 M S,? , H130,Cl E86 1.32 M Cl B&S74 1.11 M S,N,H45,Cl B&C72 0.195 P S96 0.160 M S,? NO3,Cl,SO4 St81 0.470 M SR12h,? ,H104,NO3 B78 0.810 M S96 0.290 M S96 170.0 P 0.004 M S96 0.010 M S,N,H45,Cl B&C72 0.005 M S96 110.0 P B&C72 0.003 P S96 0.040 P EC50÷ChV=4

ACR = acute-to-chronic ratio; Ch = chronic; ChV = chronic value; CC = concern concentration; ELS = early life stage; FHM = fathead m i nnow; H = hardness as CaCO 3 ; N = nom inal concentrations; ? = m e thod unknown; RT = rainbow trout; S = static m e thod; SR12h = static renewal m e thod with renewals every 12 hours; and

52W Q = water quality value.
Predictions were based on SARs for inorganic Co com pounds; pH 7; hardness <180.0 m g /L as CaCO 3 ; 100% active ingredients; m ean m easured concentrations of Co; and total organic carbon (TOC) <2.0 m g /L; Environm ental concerns for Co com pounds:

m oderate concern for acute toxicity to fish; m oderate concern for acute toxicity to daphnids; high concern for toxicity to green algae; m oderate concern for chronic toxicity to fish; high concern for chronic toxicity to daphnids; high concern for toxicity to green algae;

assessm ent factor (AsF) = 10.0 COC for fish = 0.030 COC for daphnids = 0.001 COC for green algae = 0.004 The aquatic toxicity for the chelates and esters of Co are expected to be less than predicted using toxicity data based on the soluble ion and with a MW adjustm e nt. The aquatic toxicity for organoCo could be higher than predicted using toxicity data based on the soluble ion and with a MW adjustm e nt because the organic portion could enhance uptake in aquatic organism s. It is assum e d that (1) the MW s of organoCo com pounds would have to be less than 1000 daltons and (2) the log K owof any organoCo would have to be less than 8.0 for toxicity to aquatic organism s to occur. It is also assum e d that the aquatic toxicity of organoCo com pounds will increase with increasing K owwhen the log K ow<8.0. The m a jor environm ental toxicity concerns for this category are for chronic toxicity towards daphnids and toxicity towards green algae. The acute-to-chronic ratios of Co towards fish and daphnids are both greater than 100. The SAR m e thod for Co com pounds is the nearest analog(s) m e thod. Boundaries. Cobalt com pounds m u st have water solubilities equal to or greater than 1.0 

### : g/L (ppb) and m o lecular weights generally need to be near or below 1000. The log K com pounds has to be <8.0. owof organoCo

53Fate: General Testing Strategy : I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity test s for fish (OPPTS 850.1075) and daphnids (OPPTS 850.1010) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; m easured TOC of dilution water in the control; ideally, m e an m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; and solvent can be used to assist the test chem ical to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the test chem ical beyond its aqueous solubility lim it. Stock solutions in water should be adjusted to pH near 7.0. The algal toxicity testing (OPPTS 850.5400), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; and solvent can be used to assist the test chem ical to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the test chem ical beyond its aqueous solubility lim it. Stock solutions in water should be adjusted to pH near 7.0. If there is no significant risk from the test chem ical after the results of the environm ental base set have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. For organoCo com pounds, or chelates or esters of Co, aerobic biodegradability according to either of the following test guidelines: Ready Biodegradability OPPTS 835.3110 Sealed Vessel CO2 Production Test OPPTS 835.3120 If there is no significant risk from the test chem ical after the results of biodegradation testing have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 3. Tier 3. Fish chronic toxicity testing, i.e., fish early life stage (ELS) toxicity testing (OPPTS 850.1400), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrati ons at days 7, 14, 21, and 28; m e asured TOC of dilution water in the control; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; solvent can be used to assist the test chem ical to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance

54the water solubility of the test chem ical beyond its aqueous solubility lim it; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test because the 7-d ELS toxicity test m ay underestim ate chronic toxicity m easured by the 28-d ELS toxicity test when the Chronic Values are com p ared. Stock solutions in water should be adjusted to pH near 7.0. Daphnid chronic toxicity testing (O PPTS 850.1300), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; m easured TOC of dilution water in the control; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; solvent can be used to assist the test chem ical to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the test chem ical beyond its aqueous solubility lim it; and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test because the fish 7-d ELS toxicity test m ay underestim at e chronic toxicity m easured by the fish 28-d ELS toxicity test when the chronic values are com p ared.
II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial exposures. Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay. References. B78 = Birge. 1978. USDOE Tech Info B&C72 = Biesinger&Christensen. 1972. J Fish Res Brd Can 29:1691 B&S74 = Baudouin&Scoppa. 1974. Bull Environ Contam Toxicol 12:745 E86 = Ewell etal. 1986. Environ Toxicol Chem 5:831 S96 = Suter, GW . 1996. Environ Toxicol Chem 15:1234 St81 = Stokes. 1981. J Plant Nutrit 3:667 September, 1996.

55Category: Diaz oniums Environm ental Toxicity This category includes only arom atic diazonium s (N 

### / N + ). Aliphatic diazonium s are very explosive and are only used as synthesizing agents

. Arom atic diazonium s are used as fungicides on seeds and in soil, e.g., Fenam i nosulf. It is known that diazonium s need to be absorbed to be toxic. Test data subm itted to the Agency in the New Chem icals Program showed that diazo com pounds with a num ber average m o lecular weight (MWn ) > 1000 have no effects at saturation to fish in the acute toxicity test. Therefore, com pounds with MWn' s > 1000 will be excluded f r om this category regardless of water solubility. In the NEAT state, diazonium s are solids, i.e., salts, and are m o st are soluble in aqueous m e dia. Acute toxicity for diazonium s is assum e d to be lim ited by the octanol/water partition coef f i cient (K ow ; with a m i ssing fragm e nt for N 

### / N + ) and MW

. Above a log K owvalue of => 8.0 and/or MW = 1000, diazonium s are not expected to be toxic at saturation during 96-h e xposures. Diazonium s are expected to be sim ilar to anilines with respect to their K owcutof f value f o r acute toxicity. This assum e s that the m i ssing fragmentfor N 

### / N + = -1.0. The acute toxicity of anilines dim i nishes at about a log K ow= 7.0 (Veith and Broderius 1987). There are no known K owlim its f o r chronic toxicity at this tim e, but it m ay not be m u ch above a log K ow= 10.0 for diazonium s. Future testing will determ ine this K owlim it.
Haz a rd Concerns. The acute toxicity for diazonium s has been determ ined through SAR Analysis: log fish 96-h LC 50 (m illim oles/L) = -2.456 - 0.331 log K ow based on a log K owfor the diazonium using CLOGP Ver. 3.3 with a m i ssing fragm e nt for N 

### / N + , N = 3, and R2 = 0.98. Acute toxicity values are assum e d to be valid through log K ow= 8.0. Daphnids are assum e d to have sim ilar sensitivity to fish, but green algae are expected to be m o re sensitive based on the use of diazonium s as fungicides.
Fate : Diazonium fungicides, e.g., Fenam i nosulf, ar e sensitive to light but is stabilized with Na sulphite. Fenam i nosulf is stable in alkaline m e dia. Diazonium s are also used in photography, e.g., diazo reproduction paper and film

. Therefor e, diazonium s are expected to be subject to rapid direct and indirect photolysis under envir onm entally realistic conditions. Diazonium s are also expected to slowly hydrolyze to phenols. Boundaries. : There are no known lower boundaries. The upper boundaries will be based on K owand MW

. Acute toxicity expected with log K ow< 8.0 with a m i ssing fragm e nt for N 

### / N + .; no effects at saturation during 96-h exposures when log K ow>= 8.0. Chronic toxicity has no known upper bound for log K ow , but it is probably near 10.0 with a m i ssing fragm e nt for N 

### / N +

. MWm u st be < 1000. The environm ental base set of tests will be requested for aquatic releases and the terrestrial base set of tests will be recomm e nded for terrestrial exposures. Wh en the log K owis >= 10.0, chronic toxicity testing with fish and daphnids will be recomm e nded.

56General Testing Strategy . I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. Since diazonium s com pounds are photosensitive to uvlight, it is best to avoid the use of fluorescent lights in toxicity tests. If they are used, then glass filters should be used. The acute toxicity tests for fish (CFR §797.1400) and daphnids (CFR §797.1300) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. The algal toxicity testing (CFR §797.1050), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; the highest treatm e nt concentration on a nom inal-basis cannot exceed the PMN' s aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. If there is no significant risk from the PMN after the results of the environm ental base set have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Direct and Indirect Photolysis Screening Test (40 CFR 796.3765). If t1⁄2 < 2 days, go to Tier 3; if t1⁄2 > 2 days, go to Tier 4. Tier 3a. If t1⁄2 < 2 days and photolysis products are known and/or identified, then assess photolysis products for environm ental hazards. Tier 3b. If t1⁄2 < 2 days and photolysis products are not known and/or identifiable, then prepare a stock solution of PMN using the standard hum ic-containing solution described in the direct and indirect photolysis screening te st [40.796.3765 (b)(2) and (c)(2)], expose to sunlight for at least 6 half-lives (t1⁄2), and test photolysis products for toxicity with m o st sensitive species f r om environm ental base set. For exam ple, the m o st sensitive species f r om the environm ental base set has an EC50 value = 2.0 m g PMN/L (based on 100% active ingredients [AI]), therefore, prepare a 5.0 m g PMN per liter stock solution based on 100% AI using the standard hum iccontaining solution. This stock solution is exposed to sunlight for at least 6 half-lives to ensure that all of the PMN has been photolyzed, and then this stock solution is used to retest the m o st sensitive aquatic species to determ ine if the photolysis products of the PMN are m o re or less

57toxic that the PMN. Tier 4. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (CFR §797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the PMN; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test because Van Leeuwen et al (1990) have dem onstrated that the 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the 28- d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen); Daphnid chronic toxicity testing (C FR §797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; m easured TOC of dilution water in the control; the highest treatm ent concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test because Van Leeuwen et al (1990) have dem ons trated that the fish 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the fish 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen). Aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Aerobic Aquatic Biodegradation 40 CFR 796.3100 Modified Sturm Test 40 CFR 796.3260 Closed Bottle Test 40 CFR 796.3200 Modified OECD Screening Test 40 CFR 796.3240 Modified MITI Test (I) 40 CFR 796.3220 Modified AFNOR Test 40 CFR 796.3180
II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test, the avian acute oral toxicity test, and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial exposures. Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, avian reproduction toxicity test, and the soil m i crobial comm unity bioassay.

58References. Van Leeuwen CJ, Adem a DMM, and Herm ens J. 1990. Quantitative structure-activity relationships for fish early life stag e toxicity. Aquatic Toxicology 16:321-334. Veith GD and Broderius SJ. 1987. Structure-toxi city relationships for industrial chem icals causing type (II) narcosis syndrom e. In Kaiser KLE (ed), QSAR In Environm ental Toxicology - II, p 385-391. Reidel Publishing Com p any.

Novem b er, 1991

59Category: Dichlorobenz idine-based Pigments Hum an Health Environm ental Toxicity Other Names: Diarylide Pigm ents, DCB Pigm ents, Pigm ent Yellows Definition: Any diazo pigm ent containing the substructure, dichlorobenzidine, and coupled with acetoacetanilide.

Haz a rd Concerns. There are oncogenicity/m utagenicity concerns for dichlorobenzidine-based pigm ents based on the potential release of 3,3' -dichlorobenzidine and on the presence of residual (unbound) dichlorobenzidine. DCB is a known anim al carcinogen and a suspect hum an carcinogen. In addition, DCB is known to bioconcentrate in the tissues of aquatic organism s. Boundaries. Concern for the intact pigm ent is restricted to uses at tem p eratures exceeding 200 o C. Data subm itted to the Agency under TSCA section 8(e) show that DCB pigm ents break down to release DCB as a vapor from colored polym ers when heated to extrusion tem p eratures (> 200 o C) and, from sheetm e tal coatings during curing. Though little inform ation exists on the biodegradation of pigm ents in sedim e nts, data on other low water soluble colorants, indicate that biodegradation m ay occur over a period of m onths, possibly resulting in the release of DCB. General Testing Strategy. EPA' s New Chem icals Program considers the follo wing tests to be appropriate to address the potential for DCB pigm ents to pose a significant risk to health or the environm ent: 1. Monitoring data to detect the presence of DCB under actual conditions of use; tem p erature, dwell tim e, % pigm ent in polym er or coating, and type of polym er or coating. 2. If there are releases to water, an anaerobic biodegradation assay.

60References: Appleton HT, Sikka HC. 1980. Accum u lation, elim ination, and m e tabolism of 3,3'

- dichlorobenzidine in the bluegill sunfish. Environ Sci Technol 14:50-54. Pliss GB. 1963. On som e regular relationship between carcinogenicity of am inobiphenyl derivatives and the structure of the substance. Acta Unio Int Cancrum 19:499-501. Stula EF, Sherm an H, Reinhardt CF. 1975. Expe rim e ntal neoplasia in rats from oral adm i nistration of 3,3' -dichlorobenzidine, 4,4' -m ethylene-bis(2-chloroaniline), and 4,4'

- m e thylene-bis(2-m e thylaniline). Toxicol Appl Pharm acol 32:159-176. Stula EF, Barnes JR, Sherm an H. 1978. Liver and urinary bladder tum o rs in dogs from 3,3- dichlorobenzidine. 1978. J Environ Pathol Toxicol 1:475-490. TSCA Section 8(e) Docum e nts: 8EHQ-0490-0962 INIT 8EHQ-0590-0962 SUPPL 8EHQ-0690-0962 FLW P 8EHQ-0790-0962 SUPPL 8EHQ-0890-0962 SUPPL

March, 1994

61Category: Dithiocarbamates Environm ental Toxicity This category includes N,N-dialkyldithiocarbam a tes (DDC); ethylenebisdithiocarbam a tes (BDC); and their m e tal salts which include but are not lim ited to zinc, sodium , iron, m anganese, copper, lead, m e rcury, silver and selenium

. The alkyl groups of the DDCs generally include, m e thyl through butyl, but m ay be larger. This cat egory also includes the degradation products of DDC and BDC which m ay include disulf ide m o ieties, sulf ide m o ieties, thiourea m o ieties, urea m o ieties, polym eric sulf ide m o ieties, dithizaole-3-thiones, cyclic thioureas and cyclic ureas as indicated in the generic environm ental hazard assessm ent.

Haz a rd Concerns: Many m e m b ers of this category are comm e rcial insecticides, fungicides, disinfectants, rodenticides, antioxidants, slim icides, algalicides, bactericides and heavy m e tal chelators. Their m ode of toxic action apparently results from interference with m e tallo-enzym es in living cells; the toxicity has been attributed to either DDCs and BDCs or their degradation products. All of the known dithiocarbam a tes are acutely toxic to fish, algae and bacteria at < 10 m g /L, and to aquatic invertebrates at < 1 m g /L. Chronic toxicity to fish and aquatic invertebrates ranges from 0.001 to 2 m g /L and 0.011 to 0.111 m g /L, respectively. In general, the SARs for the dithiocarbam a tes and their degradation products are sigm oidal with acute and chronic toxicity increasing with increasing Kow. The sigm oidal relationship between Kow and toxicity of the dithiocarbam a tes is very poor statistically. Consequently, toxicity predictions will be m a de using either the closest analog or averaging data for the two closest analogs which bracket the dithiocarbam a te under question. The SAR for the degradation products is m u ch m o re robust and a series of SARs will be used to predict acute toxicity of degradation products toward fish, daphnids and Photobacterim phosphoreum ; and chr onic toxicity toward fish, daphnids and green algae. Boundaries: There are no known lower boundaries. The upper boundaries are based on K and MW

. Wh en the log Kow value is < 5 m g /L, the environm ental base set of tests will be requested f o r aquatic releases and the terrestrial base set of tests will be recomm e nded f o r ow

62terrestrial exposures. Wh en the log K owis between 5 and < 19, only chronic toxicity testing will be recomm e nded. Wh en the log K owis > 19 (CLOGP), no testing will be requested because no toxic ef f ects at saturation will be expected. Generally, m e m b ers of this category will have MW s of less than 1000. General Testing Strategy Tier 1. The acute aquatic base set of environm ental toxicity tests will be recomm e nded for aquatic exposures and the terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm acute toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial exposures. Tier 2. If acute toxicity testing indicates a si gnificant risk, then chronic toxicity with fish and aquatic invertebrates will be recomm e nded as well as aerobic biodegradation testing. Aerobic biodegradability can be determ ined using one of the following test guidelines, listed in approxim a te order of preference: Aerobic aquatic biodegradation Modified Sturm test Closed bottle test Modified OECD screening test Modified MITI test (I) Modified AFNOR test
August 1989; revised Novem b er 1995 40 CFR 796.3100 40 CFR 796.3260 40 CFR 796.3200 40 CFR 796.3240 40 CFR 796.3220 40 CFR 796.3180

63Category: Epoxides Hum an Health Environm ental Toxicity Definition. Any m o lecular structure containing one or m o re epoxy groups is considered to be a m e m b er of the category:
O / \ -C---C­

| |
Haz a rd Concerns. Health concerns for epoxides are for cancer and reproductive effects based on data for several analogous chem icals. There is greater concern for prim ary epoxides,
O / \ -C---CH

H H than for epoxides with substitutions on both of the epoxy carbons. Environm ental toxicity is a function of the octanol-water partition coefficient. Com pounds with log P' s >5 act as neutral organics producing sim p le narcosis, but at lower log P' s, epoxides display toxicity greater than that predicted f o r sim p le narcotics. A QSAR (quantitative structure-activity relationship) to predict the environm ental toxicity of epoxides is under developm ent. Boundaries. Structures with epoxy equivalent weights of > 1,000 are presum ed not to pose a hazard under any conditions. Concerns are confined to those species with m o lecular weights <1,000. Health concerns are restricted to species with m o lecular weights <500 if exposure is lim ited to the derm al route. Testing. To address health concerns the following tests are usually recomm e nded for m e m b ers of this class: (1) a lifetim e cancer bioassay by the expected route of exposure, and (2) a 90-day subchronic with attention to pathology of the reproductive organs. To address ecotoxicity concerns, base set acute aquatic toxicity testing (algae: static m e thod, daphnid and fish: flowthrough m e thod, all m easured concentrations). Septem ber, 1988

64Category: Esters Environm ental Toxicity This category includes all esters, polyesters, vinyl esters, allylic esters, propargylic esters, aliphatic esters, arom atic esters, carboxylic acid esters, and sulfonate esters. These com pounds need to be absorbed to be toxic, therefore, com pounds with MW s > 1000 will be excluded from this category. Acute toxicity for esters which are liquids at room tem p erature is known to be lim ited by the octanol/water partition coef f i cient (Kow ). Above a log K owvalue of => 5.0, esters show no effects at saturation during 96-h exposures (Veith et al 1984). Esters which are solids at room tem p erature m ay show no toxicity at saturation at lower K owvalues depending on the m e lting point, i.e., the higher the m e lting point at a given K ow , the greater the likelihood that no acute toxicity will be observed at saturation. Fo r solids, the no-effects-at-saturation point has to be determ ined on a case-by-case basis. The K owlim it f o r chronic toxicity is set at a log K ow= 8 f o r liquid esters. For solid esters, chronic toxicity testing will determ ine this K owlim it.
Haz a rd Concerns. The toxicity for sim p le esters has been determ ined through SAR Analysis (Clem e nts 1988). Esters are known to be m o re toxic than neutral organic chem icals, and this excess toxicity decreases with increasing K ow . The toxicity for vinyl esters, allylic esters, and propargylic esters is expected to be greater than for sim p le esters. Again, the additional excess toxicity of these vinyl esters, allylic esters, and propargylic esters is expected to decrease with increasing K ow . Mem b ers of this category exhibit toxicity rangi ng from low toxicity (i.e., > 100 m g /L) to high

ow , MW , and m e lting point. toxicity (i.e., < 1 m g /L) depending on their K Boundaries

. There are no known lower boundaries. The upper boundaries will be based on Kow and MW

. Acute toxicity is expected when log K ow< 5.0; no effects at saturation during 96-h exposures when log K ow> 5.0. The upper boundary for chronic toxicity is 8.0. MW will be < 1000. The environm ental base set of tests will be requested for aquatic releases and the terrestrial base set of tests will be recomm e nded for terrestrial exposures. Wh en the log K owis > 5.0, chronic toxicity testing with fish and daphnids will be recomm e nded.
Fate : Esters are subject to both abiotic and biotic hydrolysis, i.e., ester hydrolysis, and aerobic biodegradation. Aerobic biodegradation is expected to be the dom inant route of transform a tion in the environm ent. General Testing Strategy

. I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity tests for fish (40 CFR §797.1400) and daphnids (40 CFR §797.1300) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it; and solvent can be used to assist the ester to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the ester
65beyond its aqueous solubility lim it. The algal toxicity test (40 CFR §797.1050) should be done with the static m e thod; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the ester; and solvent can be used to assist the ester to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the ester beyond its aqueous solubility lim it. If there is no significant risk from the ester after the results of the environm ental base set have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Aerobic Aquatic Biodegradation 40 CFR 796.3100 Modified Sturm Test 40 CFR 796.3260 Closed Bottle Test 40 CFR 796.3200 Modified OECD Screening Test 40 CFR 796.3240 Modified MITI Test (I) 40 CFR 796.3220 Modified AFNOR Test 40 CFR 796.3180 If there is no significant risk from the ester after the results of the aerobic biodegradation testing have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 3. Tier 3. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (40 CFR §797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested chem ical; solvent can be used to assist the ester to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the ester beyond its aqueous solubility lim it; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test because Van Leeuwen et al (1990) have dem onstrated that the 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen). Both anilines and esters are m o re toxic than predicted based on narcosis alone, i.e., both esters and anilines have excess toxicity due to a m o re specific m ode(s) of toxic action. A seven day exposure m ay not allow enough tim e for this excess toxicity to be expressed either because of not enough exposure and/or not enough tim e for m e tabolic activation. Daphnid chronic toxicity testing ( 40 CFR §797.1330), with flow-through m e thods;
66m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the ester; solvent can be used to assist the ester to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the ester above its aqueous solubility lim it; and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test (Van Leeuwen et al 1990).
II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test, the avian acute oral toxicity test, and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial exposures. Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, the avian reproductive toxicity test, and the soil m i crobial comm unity bioassay. References. Clem ents, RG (editor). 1988. Estim ating toxicity of industrial chem icals to aquatic organism s using structure-activity relationships. EP A-560-6-88-001. W a shington, DC: Environm ental Effects Branch, Health and Environm ental Revi ew Division, Office of Toxic Substances (TS796), United States Environm ental Protection Ag ency. Available from the National Technical Inform ation Service, Springfield, VA 22161, PB89-117592. Van Leeuwen CJ, Adem a DMM, and Herm ens J. 1990. Quantitative structure-activity relationships for fish early life stag e toxicity. Aquatic Toxicology 16:321-334. Veith GD, DeFoe D, and Knuth M. 1984. Structur e-activity relationships for screening organic chem icals for potential ecotoxicity effect s. Drug Metabolism Reviews 15(7):1295-1003.
Novem b er, 1991

67Category: Ethylene Glycol Ethers Hum an Health Definition. The ethylene glycol ether category is defined as follows: R-(OCH 2 CH 2 ) n -OR' n = 1, 2, or 3 R = alkyl C 7 or less or phenyl or alkyl substituted phenyl
R' = H or alkyl C 7 or less or any group that can be chem ically or m e tabolically rem oved to yield a glycol ether
Haz a rd Concerns. Short-chain ethylene glycol ethers are absorbed by all routes of exposure and have caused irritation of skin, eyes, and m u cous m e m b ranes; hem o lysis, bone-m arrow dam a ge, and leukopenia of both lym phocytes and granulocytes; direct and indirect kidney dam a ge; liver dam a ge, imm unotoxicity, and central nervous system (CNS) depression. Shortchain ethylene glycol ethers are also developm ental and reproductive toxicants. 2- Phenoxyethanol is known to cause hem o lysis and eye irritation. Boundaries. There is evidence that developm ental toxicity is reduced going from the m e thyl to the butyl ether, and that it is reduced going from the ethylene glycol to the triethylene glycol. However, there is still a concern f o r m a ternal toxicity as ref l ected in developm ental and subchronic toxicity studies. The system ic toxic ity of longer-chain glycol ethers and alkylphenyl glycol ethers is uncertain because data are not available. The alkyl chain length of C 7 or less was chosen as a boundary for short-chain ethylene glycol ethers based on the available data. General Testing Strategy The New Chem icals Program considers the following tests to be the m o st appropriate for ethylene glycol ethers with sufficient exposure to potentially pose an unreasonable risk: Tier 1 - Com b ined Repeated Dose Toxicity Study with the Reproduction/Developm ental Toxicity Screen Test (OECD Guideline 422). If signs of hem a turia are seen red and white blood cell counts should be taken 2 days later except for fem a le anim als during pregnancy and lactation. Tier 2 - The need for further testing would be determ ined by the results of Tier 1. This could include any of the following tests. Prenatal Developm ental Toxicity via the m o st appropriate route (40 CFR 799.9370) 2-Generation Reproduction Study via the m o st appropriate route (40 CFR 799.9380) 90-Day Subchronic Study via the m o st appropr iate route (40 CFR 799.9346 - inhalation route; 870.3250 - derm al route; 870.3150 - oral route)

68Imm unotoxicity Study via the m o st appropriate route (OPPTS 870.7800) June 1992, revised Decem ber 1997

69Category: Hydraz ines and Related Compounds Hum an Health Environm ental Toxicity

Definition: Any structure containing one or m o re of the following groups is considered to be a m e m b er of the category:

Haz a rd Concerns: Concerns for carcinogenicity and chronic effects to liver, kidney, and blood are based on data for a num ber of hydrazines and related chem icals. In hum ans, hydrazine, itself, m ay affect the central nervous system , liver, and kidneys. The toxic effects of hydrazine exposure to hum ans m ay range from m ild skin and eye irritation, and skin sensitization, to severe irritation and burns, pulm onary edem a, CNS depression, as well as liver and kidney dam a ge, which can lead to death. Hydrazine m ay also present a serious hazard to plant life and aquatic organism s. Ecotoxicity concerns are based on structure activity relationships (SAR) using data for a num ber of hydrazines and hydrazides. Hydrazine itself (N 2 H 4 ), is known to be acutely toxic to aquatic organism s at low levels, algae at < 100 ppb, 200 ppb for fish, and 30 ppb for daphnids.

Boundaries: There are no established boundaries for this category. The "typical" new chem ical m e m b er of the category is a discrete (class I) chem ical with a m o lecular weight <500. There is a greater concern for chem icals with few substitutions on the functional group than for

70those with m u ltiple substitutions.
General Testing Strategy EPA considers the following tests to be appropriate to address health and ecotoxicity concerns: 1. Lifetim e cancer bioassay by the expected route of exposure in two species of rodents (40 CFR 798.3300). 2. 90-Day subchronic in one species of rodent by the expected route of exposure to assess effects to the liver, kidney, and blood (40 CFR 798.2650). 3. Base-set ecotoxicity testing to include fish (40 CFR 797.1400) using the flow-through m e thod, daphnids (40 CFR 797.1300) using the flow-through m e thod and algae (40 CFR 797.1050) using the static m e thod, all m easured concentrations. Results of the acute ecotoxicity testing m ay trigger chronic fish (40 CFR 797.1600) and daphnid (40 CFR 797.1350) testing. 4. Environm ental fate testing including, as appropriate, m e lting point (40 CFR 796.1300) or boiling point (40 CFR 796.1220), water solubility (40 CFR 796.1840 or 796.1860), log K ow(40 CFR 796.1550, 796.1570 or 796.1720), vapor pressure (40 CFR 796.1950) and hydrolysis (40 CFR 796.3500). For arom atic hydrazines and related com pounds, the following additional testing is recomm e nded; direct and indi rect photolysis (40 CFR 796.3765), and aerobic biodegradation. Aerobic biodegradability can be determ ined using one of the following test guidelines, listed in approxim a te order of preference: Aerobic aquatic biodegradation Modified Sturm test Closed bottle test Modified OECD screening test Modified MITI test (I) Modified AFNOR test

References. 40 CFR 796.3100 40 CFR 796.3260 40 CFR 796.3200 40 CFR 796.3240 40 CFR 796.3220 40 CFR 796.3180

1. Bhide, S.V., R.A. D' Souza, M.M. Sawai, & K.J. Ranadive (1976). International Journal of Cancer 18: 530. 2. Biancifiori, C (1970). Journal of the National Cancer Institute 44: 943.

713. Biancifiori, C (1971). Lav. 1 st Anat. Istol. Pat., Univ. Studi Perugia 31: 5. 4. Biancifiori, C., E. Bucciarelli, D.B. Clayson, & F.E. Santilli (1964). British Journal of Cancer 18: 543.
5. Hydrazine-RM2 Exit Docum e nt. OPPT Office Director' s Meeting, Monday, Decem b er 13, 1993. 6. Juhasz, J., J. Balo, & B. Szende (1966). Nature (London) 210: 1377.
7. Juhasz, J., J. Balo, & B. Szende (1967). Z. Krebsforsch 70: 150. 8. Toth, B. (1969). Journal of the National Cancer Institute 42: 469. 9. Toth, B. (1972). International Journal of Cancer 9: 109. Septem ber, 1988; revised June, 1994, revised October 1995.

72Category: Hindered Amines Hum an Health Definition. The category is at present not well de fined. A "typical" new chem ical hindered am ine of concern has two or m o re hindered am ine functional groups, usually the 2,2,6,6- tetram ethyl-4-piperidinyl group, and is used as a UV light stabilizer.

Haz a rd Concerns. Health concerns f o r the category are based on data subm itted to the Agency under §8(e) of TSCA for Tinuvin 144 and Chim assorb 944. The data indicate that these hindered am ines, and presum ably hindered am ines sim ilar in structure, are toxic to the imm une system , liver, blood, the m a le reproductive system , and the G.I. tract. Boundaries. The boundaries of the category are not well defined. Tinuvin 144 has a m o lecular weight of 685, whereas Chim assorb 944 is a polym er with a num ber average m o lecular weight well in excess of 1,000. As a consequence, there is at present no m o lecular weight cutoff for hindered am ines of concern to the new chem ical program

. It is assum e d that there is little or no derm al absorption of problem atic hindered am ines because of their high m o lecular weights. Consequently, hindered am ines in the new chem ical program are only of concern if there is signifi cant inhalation exposure associated with their m anufacture, processing, or use. General Testing Strategy For hindered am ines found to pose a potentially unreasonable risk, a 90-day oral subchronic test in rats is the recomm e nded test. W e have requested that em phasis be placed on hem a tology, the imm une system , and on the m a le reproductive system

. June 1990; revised March 1995

73Category: Imides Environm ental Toxicity This category includes all im ides and m a leim ides. Substitutions m ay be aliphatic, arom atic, and/or halogens. The m ode of toxic action of im ides is unknown, but halogenated im ides are used as m i crobial pesticides, specifically, fungicides, bactericides, slim icides, and algicides. It is assum e d that these com pounds need to be absorbed to be toxic, therefore, com pounds with MW s > 1000 will be excluded from this category. Acute toxicity for im ides which are liquids at room tem p erature is assum e d to be lim ited by the octanol/water partition coef f i cient (Kow ), and the lim iting K owvalue for acute toxicity is assum e d to be about 5.0. The lim iting value for chronic toxicity is assum e d to be about 8.0. Im ides which are solids at room tem p erature m ay show no toxicity at saturation at log K owvalues < 5.0 depending on the m e lting point, i.e., the higher the m e lting point at a given K ow , the greater the likelihood that no toxicity will be observed at saturation. For solids, the no-effects-at-saturation determ ination has to be m a de on a case-bycase basis.
Haz a rd Concerns. The acute toxicity f o r im ides and m a leim ides towards f i sh has been determ ined through SAR Analysis by EPA. The SAR for acute toxicity to fish is defined by the following regression equation: log 96-h LC50 = 1.256 - 0.76 log K ow where the LC50 is in m illiMoles per Liter (m M/L), N = 4, and R 2 = 0.98. The acute toxicity of im ides towards daphnids is expected to be sim ilar to that of fish, but their toxicity towards green algae is expected to be greater because of their use as m i crobial pesticides. Polyim ides m ay be m o re toxic than predicted using this SAR. The toxicity of im ides towards aquatic organism s can range from low (i.e., > 100.0 m g /L) to

owand MW

. The higher the K lower the MW , the higher the toxicity (or the lower the EC50 value). high toxicity (i.e., < 1.0 m g /L) depending on their K owand the

Boundaries. : There are no known lower log K owand MW boundaries. The upper boundaries f o r acute toxicity will be set at a log K ow< 5.0; chronic toxicity lim its will be set at a log K ow< 8.0. MW will be < 1000 for stable com pounds. The environm ental base set of tests will be requested f o r aquatic releases and the terrestrial base set of tests will be recomm e nded f o r terrestrial exposures. General Testing Strategy. The aquatic base set of environm ental toxicity tests will be recomm e nded for aquatic exposures. The acute toxicity tests for fish (40 CFR 797.1400) and daphnids (40 CFR 797.1300) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis

74should equal the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility. The algal toxicity testing (40 CFR 797.1050), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; the highest treatm e nt concentration on a nom inal-basis equal to the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artif icially enhance the water solubility. Fish chronic toxicity testing, i.e., fish ear ly life stage (ELS) toxicity testing (40 CFR 797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test because Van Leeuwen et al (1990) have dem onstrated that the 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen); Daphnid chronic toxicity testing ( 40 CFR 797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; m easured TOC of dilution water in the control; the highest treatm ent concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility; and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test because Van Leeuwen et al (1990) have dem onstrated that the fish 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the fish 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen). The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test [40 CFR 797.2800], the earthworm acute toxicity test [40 CFR 795.150], and the soil m i crobial comm unity bioassay [40 CFR 795.3700]) will be recomm e nded for terrestrial exposures. Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test [40 CFR 797.2830], the plant uptake test [40 CFR 797.2850], and the soil m i crobial comm unity bioassay.

75References. Van Leeuwen CJ, Adem a DMM, and Herm ens J. 1990. Quantitative structure-activity relationships for fish early life stag e toxicity. Aquatic Toxicology 16:321-334. April, 1991

76Category: Diisocyanates Hum an Health Definition. Any m o lecular structure containing two or m o re isocyanate groups is considered to be a m e m b er of the category for new chem ical purposes:
R-(N=C=O) > 2

Mem b ers of the class include new isocyanate m onom ers as well as new oligom ers, polym ers, prepolym ers, or reaction products of existing isocyanate m onom ers. Most new chem ical diisocyanates of concern are polym ers or oligom ers containing well-known diisocyanate m onom ers such as toluene diisocyanate (TDI) or 4,4' -m ethylenediphenyl diisocyanate (MDI).
Haz a rd Concerns. Diisocyanates are of concern for potential derm al and respiratory sensitization, and for pulm onary toxicity. Ba sed on conflicting anim al and hum an data for respiratory sensitization, the Agency has determ ined that there is presently not a reliable anim al m odel for testing diisocyanates for potential respiratory sensitization. At this tim e, it is assum e d that all diisocyanates m ay be potential hum an respiratory sensitizers.
Most m e m b ers of the diisocyanate category have not been tested for carcinogenic potential. Though the arom atic diisocyanates [MDI, TDI, dianisidine diisocyanate (DADI)] tested positive and one aliphatic diisocyanate [ h exam ethylene diisocyanate (HDI)] tested negative in one species, it is prem ature to m ake any generalizations about the carcinogenic potential of arom atic versus aliphatic diisocyanates. Boundaries. Structures with an isocyanate equivalent weight of > 5,000 are presum ed not to pose a hazard under any conditions. Typically, concerns are confined to those species with m o lecular weights <1,000. Frequently, new chem ical isocyanates are m anufactured with a significant excess of isocyanate m onom er. Under these circum stances, the excess m onom er is usually regarded as m o re hazardous than the "new" chem ical com ponent, and these PMN substances are ordinarily not regulated under §5 of TSCA. For the purposes of risk assessm ent within the New Chem icals Program , a PMN substance is considered "existing" if m o re than 50% of the free isocyanate groups in the PMN substance (new chem ical com ponent + existing chem ical m onom er) reside on unreacted m onom er(s). This does not relieve a Com p any, however, of any obligations to subm it a PMN for the new chem ical isocyanate if indeed it is not listed on the TSCA Inventory. General Testing Strategy

. T he following testing is recomm e nded to address the potential for pulm onary toxicity and derm al sensitization.

1. Derm al sensitization (OPPTS 870.2600).

772. 90-day Subchronic inhalation toxi city test in rodents (OPPTS 870.3465). In addition, appropriate hazard comm unication needs to be developed and im plem ented. Health and Safety Information. The following inform ation provides guidance in developing hazard comm unication and protective m easures language to accom p any new diisocyanate chem icals and form ulations. It is based on the Agency’s current understanding of the hazards associated with diisocyanates and the m o st effective m eans to lim it exposure. Warnings

. Exposure to diisocyanates m ay cause the following hum an health effects: skin irritation and allergic reactions, respiratory irritation, respiratory sensitization, and lung toxicity; som e diisocyanates also m ay cause cancer. The likelihood that these effects will occur depends on a num ber of factors; am ong them , the level of exposure, frequency of exposure, part of the body exposed, and sensitivity of the exposed individual. Sym p tom s of allergic reaction and respiratory sensitization include rashes, cough, shortness of breath, asthm a , chest tightness and other breathing dif f i culties. There is uncertainty as to the m echanism by which sensitization occurs. In sensitized individuals, exposure to even sm all am ounts of diisocyanates (below governm e nt-recomm e nded workplace exposure levels) m ay cause allergic respiratory reactions like asthm aand severe breathing dif f i culties. It is especially im portant to note that contact with skin m ay lead to respiratory sensitization or cause other allergic reactions

. In som e cases, the effects of diisocyanate exposure m ay be imm e diate and life-threatening; in others, the effects m ay be delayed and occur hours after the exposure has ended. Repeat or prolonged exposure to diisocyanates m ay also cause irritation to eyes, skin, respiratory tract and lungs, as well as adverse chronic lung effects, like decreased lung capacity and function. Individuals experiencing shortne ss of breath, tightness in the chest or other problem s breathing should seek imm e diate m e dical attention. Protective Measures. In workplaces where individuals handle diisocyanates or coatings or other form ulations that contain them , an industrial hygiene and safety program should be operative. Im portant com ponents of this program include: hazard comm unication and training on safe handling practices; use of efficient and well-m a intained application equipm ent, engineering controls and personal protective equipm ent; housekeeping procedures including spill prevention and cleanup practices; and, if feasible, m eans to m easure airborne levels of polyisocyanates and diisocyanates. During spray applications, workers should take precautions to avoid breathing vapors, m i sts or aerosols. Inhalation exposures should be lim ited to <0.05 m g /m 3 as an 8-hour tim e-weighted

78average (TWA ) for com b ined polyisocyanates and diisocyanates. 1 Engineering controls should serve as the first, m o st effective m eans of reducing airborne polyisocyanate and diisocyanate concentrations; an appropriate NIOSH/MSHA-a pproved respirator should be used as a secondary tool to lower exposures. Curren tly, downdraft spray booths and high-volum e lowpressure (HVLP) spray guns appear to offer th e m o st efficient technology to reduce inhalation exposures; a m a intenance program should always be used to ensure optim al operating efficiencies. To lim it derm al contact, indivi duals should wear im perm eable gloves, protective clothing and goggles or glasses with side shields.

May 1990, revised July 1993, February 1995, and February, 1997

1 0.05 m g /m 3 or 0.005 ppm TWA is the Am erican Conference of Governm e nt Industrial Hygienists (ACGIH) Threshold Lim it Value (TLV ) for each hexam e thylene diisocyanate (HMDI), toluene-2,4-diisocyanate (TDI), and m e thylene bi sphenyl isocyanate (MDI). Also, OSHA has set 0.02 ppm as exposure ceilings for both TDI and MDI. 79Category: 

### $ -Naphthylamines, Sulfonated Hum an Health Definition: Any new chem ical whose structure is consistent with the following general structure is considered to be a m e m b er of the category of m onosulfonated 

### $ -naphthylam ines:

Included in the category are azo dyes which release a sulfonated $ -naphthylam ine upon reduction of azo bonds. Also included in the category are N-acetylated sulfonated $

­ naphthylam ines.
Haz a rd Concerns. Based on analogy to $ -naphthylam ine per se , m e m b ers of the class are considered potential carcinogens. A num ber of m ono- and disulfonated $ -naphthylam ines are positive in the Am es assay, and som e are active in the m ouse lung adenom a assay. The presence of one or two sulfonate groups or the sulfatoethylsulfone group is likely to slow system ic uptake and enhance excretion, however, the extent of these m itigating effects is unknown. Boundaries. Concern is restricted to sulfonated $ -naphthylam ines where not m o re than two sulfonate or sulfatoethylsulfone group(s) are on the ring distal to the $ -am i no group. The Agency has sufficient data to indicate that $ -naphthylam ines where a sulfonate group is on the proxim a l ring are unlikely to be carcinogenic. General Testing Strategy . The New Chem icals Program considers the following tests to be the m o st appropriate for m onoand di-sulfonated $ -naphthylam ines found to pose an unreasonable risk:

- An Am es test or, for azo dyes, an Am es test with the Prival m odification, and - An unscheduled DNA synthesis test in rat hepatocytes. For azo dyes, it is necessary that the specific sulfonated $ -naphthylam ine in question be isolated prior to testing. For both tests, $ -naphthylam ine is to serve as an additional positive control. If the results of the genotoxicity testing indicate that the new chem ical is genotoxic, a

80two- year, two-species cancer bioassay would be required. April 1991, updated June 1999

81------------------------------------

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------------------------------------ Category: Lanthanides or Rare Earth Metals Environm ental Toxicity Definition. This category includes inorganic salts, com p lexes organic acids, and organom etallic com pounds or lanthanides or rare earth m e tals. There are 14 naturally-occurring lanthanides or rare earth m e tals:

Nam e Sym bol MW CASRN Lanthanum La 139 [7439-91-0] Cerium Ce 140 [7440-45-1] Praseodym ium Pr 141 [7440-10-0] Neodym ium Nd 144 [7440-00-8] Sam arium Sm 150 [7440-19-9] Europium Eu 152 [7440-53-1] Gadolinium Gd 157 [7440-54-2] Terbium Tb 159 [7440-27-9] Dysprosium Dy 163 [7429-91-6] Holm ium Ho 165 [7440-60-0] Erbium Er 167 [7440-52-0] Thulium Tm 169 [7440-30-4] Ytterbium Yb 173 [7440-64-4] Lutetium Lu 175 [7439-94-3]

The lanthanide rare earth m e tals are very sim ilar to each other. Their m o st im portant oxidation state is +3 for all. None are known to be essential to biological species. Across the lanthanide series from La to Lu, there is a general or steady decrease in their (1) atom ic radii, (2) covalent radii, and (3) radii of their tripositve ions due to the addition of electrons at the 4f electron shell. This decrease in radii leads to a corresponding increase in the polarizing power of their ions and in the stability of com p lexes of their ions. They all have sim ilar chem istry behaviors.
Haz a rd Concerns. The only toxicity data available for the lanthanide series are for La [7439- 91-0]. Soluble salts of La are known to have high chronic toxicity towards fish, m oderate chronic toxicity towards green algae, and low acute toxicity towards daphnids based on exposures in m oderately hard water and in term s of m g La/L. Toxicity inform ation are only available for La trichloride [10099-58-8] and La triacetate [917-70-4]. The toxicity profile for La based on available toxicity data, i.e., m easured (M) and predicted (P), m g La/L (ppm La), and m oderate hardness (i.e., <180.0 m g /L as CaCO 3 ) is:

82fish 96-h LC50 < 1.0 P ACR10 daphnid 48-h LC50 = 160.0 M S,N BK59 green algal 96-h EC50 < 1.0 P EC50/ChV=4 fish chronic value(ChV) = 0.020 M SR12,M H104 B78 daphnid ChV = 20.0 P ACR10 algal ChV = 0.150 M S,N BK59

The lanthanides are assum e d to be m o re toxic in soft water than hard water based on data for other heavy m e tals. Boundaries. The toxicity of the lanthanide rare earth m e tals depends on the their physical/chem i cal properties and the hardness of receiving waters. The toxicity of their salts and their com p lexes with organic acids are expected to be related to their water solubility, their MW s, and the stability of their com p lexes. The toxicity of lanthanide organom etallic com pounds, if they exist, are expected to be related to their octanol/water partition coefficient (Kow). The m o st im portant property determ ining the toxic ity of chem icals derived from lanthanide rare earth m e tals is their water solubility. W a ter solubility cannot be estim ated accurately and has to be m easured. The water solubility of organom etallic com pounds is expected to decrease as Kow increases. There is no lower bound for Kow and the upper bound cannot be determ ined at this tim e since the Kow fragm e nt-constant for any of the lanthanide rare earth m e tals are not known. In addition to solubility, MW is also an im portant boundary. Highly stable com p lexes and organom etallics with MW s > 1000 are not expected to be absorbed by aquatic organism s even if they are water soluble. Therefore, only lanthanide rare earth m e tal-com pounds with MW s < 1000 are expected to be toxic. General Testing Strategy.
I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity tests for fish (40 CFR §797.1400 or OPPTS 850.1075) and daphnids (40 CFR §797.1300 or OPPTS 850.1010) will be done using the flow-through m e thod; effective concentrations will be based on 100% active ingredients (ai) and m ean m e asured concentrations; the total organic carbon (TOC) concentration of dilution water in the control m

u st be less than 2.0 m g TOC/L; TOC m u st be m easured in the control just prior to the start of the test; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested com pound; solvent can be used to assist the com pound to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the com pound beyond its aqueous solubility lim it; and hardness of dilution water has to be less than 180.0 m g /L as CaCO 3 . If toxicity m itigation testing is done with hum ic acid, then the static m e thod with nom inal concentrations will be recomm e nded.

83The algal toxicity testing (40 CFR §797.1050 or OPPTS 850.5400), should be done with the static m e thod; effective concentrations based on 100% ai and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with no m o re than 0.300 m g /L EDTA as a final concentration; the TOC of the test/growth m e dium should be less than 2.0 m g TOC/L; TOC should be m easured just prior to the start of the test; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested com pound; and solvent can be used to assist the com pound to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the com pound beyond its aqueous solubility lim it. If toxicity m itigation testing is done with hum ic acid, then nom inal concentrations will be recomm e nded. If there is no significant risk from the chem ical after the results of the environm ental base set have been integrated into the risk assessm ent, then no f urther testing will be recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (40 CFR §797.1600 or OPPTS 850.1400), with the flow-thr ough m e thod; effective concentrations based on 100% ai and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; the TOC of dilution water in the control should be less than 2.0 m g TOC/L; TOC should be m easured in the controls just prior to and during the test; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested com pound; solvent can be used to assist the com pound to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the com pound beyond its aqueous solubility lim it; and hardness of dilution water has to be less than 180.0 m g /L as CaCO 3 . Daphnid chronic toxicity testing (40 CFR §797.1330 or OPPTS 850.1300), with the flowthrough m e thod; effective concentrations based on 100% AI and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; the TOC of dilution water in the control should not exceed 2.0 m g TOC/L; TOC m u st be m easured in the controls just prior to and during the test; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested com pound; solvent can be used to assist the com pound to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the com pound beyond its aqueous solubility lim it; and hardness of dilution water has to be less than 180.0 m g /L as CaCO 3 . II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests will be recomm e nded for terrestrial exposures. The terrestrial base set includes: the early seeding growth test (OPPTS 850.4230), the earthworm toxicity test (OPPTS 850.6200), the soil m i crobial comm unity bioassay (OPPTS 850.5100), and th e avian acute oral toxicity test (OPPTS 850.2100). Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test (OPPTS 850.4150), the plant uptake test (OPPTS 850.4800), and the avian reproduction test (OPPTS 850.2300 for bobwhite quail or for m a llard duck).

84February, 1998

85Category: Neutral Organics Environm ental Toxicity

Definition. This broad category includes non-reactive non-ionizable organic chem icals such as alcohols, ketones, ethers, alkyl halides, aryl halides, and arom atic hydrocarbons.

Haz a rd Concerns. Neutral organics are environm entally toxic because of their ability to produce sim p le narcosis in aquatic species. Toxicity is a f unction of the octanol-water partition coefficient. Com pounds with log P' s of <5 e xhibit toxicity within 96 hours. At log P 5-8, toxicity is apparent only after extended exposure. Com pounds with a log P >8 are not toxic at water saturation even after prolonged exposure. There are a num ber of QSARs (quantitative structure-activity relationships) to predict the toxicity of neutral organics. Boundaries. The m o lecular weights of neutral organics of concern are generally less than 1,000. Log P is <8. QSAR predictions of toxic ity are constrained by water solubility. If a predicted toxicity level exceeds water saturation, then a longer test is needed to observe toxicity. General Testing Strategy To address ecotoxicity concerns, base set acute aquatic toxicity testing (algae (40 CFR 797.1050): static m e thod, daphnid (40 CFR 797.1300) and fish (40 CFR 797.1400): flowthrough m e thod, all m easured concentrations).

86To properly assess any hum an and environm ental toxicity or exposure, certain environm ental fate properties, such as aerobic biodegradation, need to be m easured. Aerobic biodegradability can be determ ined using one of the following test guidelines, listed in approxim a te order of preference: Aerobic aquatic biodegradation Modified Sturm test Closed bottle test Modified OECD screening test Modified MITI test (I) Modified AFNOR test 40 CFR 796.3100 40 CFR 796.3260 40 CFR 796.3200 40 CFR 796.3240 40 CFR 796.3220 40 CFR 796.3180 The physical state and electronic charge of the PMN substance should also be reported.
For som e neutral organics (e.g. ketones) dir ect/indirect photolysis (40 CFR 796.3765 and 796.3700) AND possibly biodegradation testing is recomm e nded.
For som e neutral organics (e.g. alkyl halides) hydrolysis testing (40 CFR 796.3510) AND possibly biodegradation testing is recomm e nded.
Septem ber 1988; revised October 1995

87Category: Nickel Compounds Hum an Health Environm ental Toxicity Definition. Inorganic and organic com pounds of nickel in which there is the potential for uptake of either Ni 2+ or organonickel.
Haz a rd Concerns. Nickel com pounds e.g., nickel refinery dust, and its m a jor com ponent nickel subsulfide, have been shown to be carcinogenic in hum ans. Som e nickel com pounds are known to be genotoxic. IRIS has established an oral RfD for the soluble salts of nickel of 2 x 10 -2 m g /kg/d (1992) based on effects on organ weights ina two-year feeding study in rats. In the study, there was a statistically significant reduction in total body weight, higher heart-to-body weight ratios and lower liver-to-body weight ratios than controls. In addition to the effects on organ weights found in the critical two-year study, two other sensitive endpoints exist, neonatal m o rtality and derm atotoxicity. Wh ile no reproductive effects have been associated with nickel exposure to hum ans, several studies in laboratory anim als have dem onstrated fetotoxicity. Soluble inorganic nickel com pounds produce acute and chronic toxicity in freshwater and saltwater aquatic organism s over a wide range of concentrations but bioconcentrates only to a sm all degree. There are no known toxicity data for organonickel com pounds. Boundaries. Any nickel com pound that will release Ni 2+ is considered hazardous. Conversely, there are no available data to suggest that nickel com pounds in which the Ni 2+ is not released m ay pose a health hazard. The boundaries for ecotoxicity of Ni 2+ com pounds depend on whether they are Ni 2+ salts, Ni 2+ chelates, or organonickel com pounds. There is also a m o lecular weight boundary for strong ion pairs/com p lexes which is 1000. The boundaries for organonickel com pounds (e.g., K ow undefined but the m o lecular weight boundary is expected to be 1000. of the organic portion) are
Occupational Exposure Controls. Because nickel com pounds have been shown to be toxic by the inhalation/ingestion route, exposure controls that m a intain airborne exposures at 0.1 m g /m 3 or below are needed (consistent with OSHA PEL TWA). In addition, since nickel com pounds are also toxic by the derm al route, NIOSH approved protective gloves are also recomm e nded. Testing

. D epending upon estim ated workplace exposures and releases to water, which will be assessed on a case-by-case basis, the following testing m ay be recomm e nded: To address health effects concerns due to the toxicity of nickel and its com pounds, the following tests m ay be recomm e nded: A 90-day subchronic study in rats by an appropriate route to assess system ic toxicity (OPPTS 870.3100 or 870.3250 or 870.3465)

88Results of the 90-day study m ay trigger a lifetim e bioassay in rats and m i ce by the inhalation route to assess potential carcinogenicity (OPPTS 870.4200) To address ecotoxicity concerns due to toxicity of nickel and its com pounds, the following base set tests m ay be recomm e nded: Acute fish toxicity test OPPTS 850.1075 Acute daphnid toxicity test OPPTS 850.1010 Green algae toxicity test OPPTS 850.5400 All tests utilize m easured concentrations. To properly assess hum an and environm ental toxicity or exposure, certain physicochem ical or environm ental fate properties need to be m easured:
W a ter solubility Octanol/water partition coef f i cient (Kow) Vapor pressure Melting point-m elting range Boiling point OPPTS 830.7840 or 830.7860 O PPTS 830.7550 or 830.7560 or 830.7570 OPPTS 830.7950 or 830.8000 OPPTS 830.7200 OPPTS 830.7220 In addition, aerobic biodegradation by one of the following m e thods: CO 2 evolution Closed bottle Modified OECD screening Modified MITI (I) DOC die-away Manom etric respirom etry

References. O PPTS 835.3110 OPPTS 835.3110 O PPTS 835.3110 OPPTS 835.3110 OPPTS 835.3110 OPPTS 835.3110

IRIS access. # 1271 (09/30/87) Nickel, soluble salts. OSHA PELs (1995-1996)
September 1996

89Category: Nonionic Surfactants Environm ental Toxicity Definition. Any neutral structure having surfactant activity is considered a m e m b er of this category. Many of these surfactants have the following types of structure:

Ethoxylate groups m ay be m i xed with or be re placed by alcohol groups. Other neutral groups e.g. propoxylates, esters, halogens, m ay also be present.
Haz a rd Concerns. Acute aquatic toxicity increases exponentially with increases in the hydrophobic chain length when the num ber of ethoxy groups or the hydrophilic com ponent is held constant. In addition, when the num ber of carbons in the hydrophobe are constant, toxicity decreases with an increasing num ber of ethoxylate groups. The aquatic toxicity of m e m b ers of the category can be predicted by structure-activity relationship (SAR). Boundaries. There are no established m o lecular weight boundaries for this category. Lim its on chain length are inherent in the SARs. Testing. To address ecotoxicity concerns, base set acute aquatic toxicity testing (algae: static m e thod, daphnid and fish: flow-through m e thod, all m easured concentrations). Tier 1. The acute aquatic base setof environm ental toxicity tests will be recomm e nded for aquatic exposures and the terrestrial base setof environm ental toxicity tests (i.e., the early seeding growth test, the earthworm acute toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded for any terrestrial exposures. Acute fish toxicity test Acute daphnid toxicity test Green algae toxicity test Early seedling growth test Earthworm acute toxicity test Soil m i crobial comm unity bioassay OPPTS 850.1075 OPPTS 850.1010 OPPTS 850.5400 OPPTS 850.4230 OPPTS 850.6200 O PPTS 850.5100 Tier 2. If acute toxicity testing indicates a significant risk, then environm ental fate testing in the form of aerobic biodegradation testing is recomm e nded. Aerobic biodegradability can be determ ined using one of the following test guidelines:

90CO 2 evolution O PPTS 835.3110 Closed bottle OPPTS 835.3110 Modified OECD screening O PPTS 835.3110 Modified MITI (I) OPPTS 835.3110 DOC die-away OPPTS 835.3110 Manom etric respirom etry OPPTS 835.3110 Tier 3. In addition, if acute toxicity testi ng indicates a significant risk, then chronic aquatic toxicity testing with f i sh and aquatic invertebrates will be recomm e nded. Fish early life stage test OPPTS 850.1400 Daphnid chronic toxicity testing O PPTS 850.1300
September 1988, revised September 1996

91Category: Organotins Hum an Health Environm ental Toxicity Definition. This category includes all m ono-, di-, tri- and tetra-alkyl or phenyl organotin com pounds, including organotin esters/oxides. The m ode of toxic action of organotins in hum ans is unknown, but they are known to affect carbonate m e tabolism and other m e tabolic processes in the brain, liver, and m u scle, as well as several enzym e s and the oxidative activity of m itochondria. It has been suggested that general sulfhydryl binding m ay be responsible for the effects seen in m amm a ls. It is assum e d that these com pounds need to be absorbed to be toxic, therefore, com pounds with MW s > 1000 and which do not transform to organotin com pounds with MW s < 1000 will be excluded from this category. Hum an health and aquatic toxicity for liquid organotins is assum e d to be lim ited by the relative octanol/water partition coef f i cient (a relative K ow is based on the com puter program , CLOGP, with C substituted f o r Sn). However, the lim iting relative K owvalue is unknown. The Agency has data for an organotin with a relative log K ow= 13.7 which still showed high toxicity towards fish and daphnids. Organotins which are solids at room tem p erature m ay show no toxicity at saturation at log K owvalues < 13.7 depending on the m e lting point, i.e., the higher the m e lting point at a given K ow , the greater the likelihood that no toxicity will be observed at saturation. For solids, the no-effects-at-saturation determ ination has to be m a de on a case-by-case basis. There are no known K owlim its f o r acute and chronic environm ental toxicity at this tim e, but it is higher than a log K ow= 13.7. Future testing w ill determ ine the K ow lim its.

Haz a rd Concerns. Tested organotins have been shown to be from m oderately irritating to corrosive to the skin and eyes. Acute oral and derm al exposures can result in system ic effects, prim arily neurotoxicity. Concerns for imm unotoxicity are based on data on dialkyltins and trialkyltins. During a 13-week oral study in rats using dioctyltin bis (2-ethylhexylthioglycolate), effects to the thym us, spleen, lym ph nodes, and bone m arrow were observed. A No-ObservableAdverse-Effect-Level (NOAEL) of 1.6 m g /kg/d wa s derived from this study. During a 17-m onth feeding study in rats using bis (tri-n-butyltin)-oxide, imm unoglobulin E (IgE) titers were reduced, as well as the activity of natural killer cells in the spleen, and there was reduced m acrophage function. A NOAEL of 0.025 m g /kg/d wa s derived from this study. Based on the imm unotoxicity end-point, an oral reference dose for chronic exposure (RfD) for tributyltin

92oxide of 3 x 10 -5 m g /kg/d (IRIS 01/01/89) has been derived. Organotins are well known neurotoxins, with the tri- and tetra-substituted tins being m o re toxic than the m ono- and disubstituted com pounds. Although m any tri-alkyltins show clear neurotoxic effects (eg. lesions in the hippocam pus), the neurotoxic potential of di-alkyltins has not been as well def i ned. Available data indicate that dibutyltin and di octyltin can produce neurotoxic effects such as reductions in brain neurotransm itter levels, alterations in spontaneous m o tor activity and hindlim b weakness. An effect level for neuroc hem i cal and behavioral changes following 3 days of oral adm i nistration of dibutyltin dilaurate was reported as 20 m g /kg. There are oncogenicity concerns for som e of the organotins based on analogy to triphenyltin hydroxide, a group B probable hum an carcinogen. Therefore, the hum an health concerns for phenyltins will be dealt with on a case-by-case basis. The acute aquatic toxicity for several subclasses of organotins has been determ ined through toxicity testing (Vighi & Calam ari, 1985; W ong et al. 1982) and structure activity relationships (SAR). Organotins have been shown to be highly toxic to green algae (W ong et al. 1982). The acute toxicity of organotins is m oderate to high towards daphnids (Vighi & Calam ari, 1985). One datum for fish has indicated high toxic ity (USEPA 1996). Organotins exhibit toxicity ranging from m oderate toxicity (i.e., > 10 m g /L) to high toxicity (i.e., < 1 m g /L) depending on their K ow, m e lting point, and MW

. The higher the K ow , the higher the toxicity (or the lower the EC 50 value). Boundaries. There are no known boundaries. The upper boundaries will be based on Kowand MW

. Acute toxicity expected with log K ow< 13.7; chronic toxicity has no known upper bound for log K ow , but it is > 13.7. MW will be < 1000 for stable com pounds. The environm ental base set of tests will be requested f o r aquatic releases and the terrestrial base set of tests will be recomm e nded for terrestrial exposures. General Testing Strategy. TIER 1. A 90-day subchronic test in rodents by the oral route with special attention to the lym phoid organs (thym us, spleen, peripheral lym ph nodes) and bone m arrow (OPPTS 870.3100). Neurotoxicitytesting to include m o tor activity, a functional observational battery and neuropathology with special attention to lesions in the hippocam pus (OPPTS 870.6200). This testing can be com b ined with the 90-day subchronic protocol cited above. The acute aquatic base setof environm ental toxicity tests will be recomm e nded f o r aquatic exposures and the terrestrial base setof environm ental toxicity tests will be recomm e nded f o r terrestrial exposures. The acute toxicity tests for fish and daphnids should be done using the flow-through m e thod with m easured concentrations. Effective concentrations should be based on 100% active ingredients (AI) and m ean m easured concentrations. Measure TOC of dilution water in the control just prior to testing. Ideally, the highest treatm e nt concentration on a m ean

93m easured concentration-basis should equal the aqueous solubility lim it. Solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to significantly enhance the water solubility. The algal toxicity testing should be done with the static m e thod and m easured concentrations. Statistical analysis of effective concentrations at 24, 48, 72, and 96 hours should be perform e d. The test m e dium should be used with at least 0.300 m g /L EDTA as a final concentration. Aquatic Base Set Acute fish toxicity test Acute daphnid toxicity test Green algae toxicity test Terrestrial Base Set Early seedling growth test Earthworm acute toxicity test Soil m i crobial comm unity bioassay TIER 2. OPPTS 850.1075 OPPTS 850.1010 OPPTS 850.5400

OPPTS 850.4230 OPPTS 850.6200 O PPTS 850.5100

If acute aquatic toxicity testing indicates a significant risk, the following environm ental fate testing is recomm e nded to determ ine aerobic biodegradabilityusing one of the following test guidelines: CO 2 evolution Closed bottle Modified OECD screening Modified MITI (I) DOC die-away Manom etric respirom etry TIER 3. O PPTS 835.3110 OPPTS 835.3110 O PPTS 835.3110 OPPTS 835.3110 OPPTS 835.3110 OPPTS 835.3110

If acute toxicity testing and environm ental fate te sting continue to indicate a significant risk, then chronic aquatic toxicitytesting with fish and aquatic invertebrates is recomm e nded with flowthrough m e thods and m easured concentrations. Effective concentrations should be based on 100% active ingredients (AI) and m ean m easured concentrations. Conduct statistical analysis of effective concentrations at days 7, 14, 21, and 28. Measure TOC of dilution water in the control just prior to testing. Ideally, the highest treatm e nt concentration on a m ean m easured concentration-basis should be equal at the aqueous solubility lim it. Solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to significantly enhance the water solubility. The 7-d fish early life stage (ELS) toxicity test cannot be substituted for the 28-d ELS toxicity test and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test because Van Leeuwen et al. (1990) have dem onstrated that the 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the 28-d

94ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen). Chronic Aquatic Toxicity Fish early life stage test OPPTS 850.1400 Daphnid chronic toxicity testing O PPTS 850.1300 References. 8E-594. Tributyltin oxide. Bis-(tri-n-butyltin)-oxide

. (1988, Decem ber). Gesellschaft Deutscher Chem iker - Advisory Comm ittee on Existing Chem ical of Environm ental Relevance (BUA). BUA Report 36. USEPA 1996. United States Environm ental Protec tion Agency (1996). The Office of Pollution Prevention andToxics (OPPT) prem anufacture notification (PMN) ECOTOX database: a confidential business inform ation (CBI) collection of environm ental toxicity data from new chem ical subm issions under Sec. 5 of the Toxic Substances Control Act (TSCA). W a shington, DC: Environm ental Effects Branch, Health and Environm ental Review Division (7403), OPPT, USEPA. OPPT contact is V. Nabholz, 202-260-1271. Van Leeuwen, C.J., Adem a, D.M.M., & Herm ens, J. (1990). Quantitative structure-activity relationships for fish early life stage toxicity. Aquatic Toxicology, 16, 321-334. Vighi, M., & Calam ari, D. (1985). QSARs for organotin com pounds on Daphniam a gna . Chem osphere, 14, 1925-1932.
W HO 1980. W o rld Health Organization Environm ental Health Criteria 15. Tin and organotin com pounds: A prelim inary review. Geneva: W HO pp. 59-62.
W ong, P.T.S., Chau, Y.K., Kram ar, O., & Bengert, G. A. (1982). Structure-toxicity relationship of tin com pounds on algae. Canadian Journal of Fisheries and Aquatic Sciences, 39, 483-488.
April 1991, revised September 1996

95Category: Peroxides Hum an Health (case-by-case) Environm ental Toxicity Definition. Any m o lecular structure containing one or m o re of the following functional groups is considered to be a m e m b er of the class: | | -C-O-O-C­

| | | -C-O-O-H

|
O 5 | -C-O-O-C­ |
O O 5 5 -C-O-O-C-

O 5 -C-O-O-H dialkyl peroxide

alkyl hydroperoxide

peroxy ester

diacyl peroxide

peroxy acid The category of peroxides is a sm all one (<0.2% of all new chem icals). The "typical" peroxide in the new chem ical program is a discrete (class I) chem ical with a m o lecular weight of <500.
Haz a rd Concerns. EPA had previously identified hazard concerns for the possible carcinogenicity of new chem icals falling into the peroxide category based on available evidence. EPA has reviewed test data developed by the Soci ety of the Plastics Industry (SPI) and others on the peroxide category of chem icals and concl udes that available inform ation does not support continued identification of peroxides as a ne w chem ical category presenting concerns for possible carcinogenicity. EPA will continue to evaluate the potential health concerns for new chem ical peroxides that are notif ied but, with this change, will not apply a category understanding as regards potential carcinogenicity. Wh ile the assessm ent of potential hum an health hazard concerns will be conducted on a case-by-case basis, this m odification does not affect EPA’s approach to environm ental hazard concerns which will continue to be identified for m e m b ers of this category. Boundaries. There are no established boundary conditions for review of peroxides. General Testing Strategy .

96The New Chem icals Program considers the following tests to be appropriate to address ecotoxicity concerns for peroxides: 1. Base-set ecotoxicity testing to incl ude fish (40 CFR 797.1400), daphnids (40 CFR 797.1300), and algae (40 CFR 797.1050). 2. Environm ental fate testing including, as appropriate, m e lting point (40 CFR 796.1300) or boiling point (40 CFR 796.1220), water solubility (40 CFR 796.1840 or 796.1860), log K ow(40 CFR 796.1550, 796.1570, or 796.1720), vapor pressure (40 CFR 796.1950), hydrolysis (40 CFR 796.3500), direct and indirect photolysis (40 CFR 796.3765), and aerobic biodegradation by anyof the following test guidelinesModified Sturm Test 40 CFR 796.3260 Closed Bottle Test 40 CFR 796.3200 Modified OECD Screening Test 40 CFR 796.3240 Modified MITI Test (I) 40 CFR 796.3220 Modified AFNOR Test 40 CFR 796.3180 Septem ber, 1988; revised June, 1994; revised April, 2001

97Category: Persistent, Bioaccumulative, Environm ental Fate and Toxic (PBT) Chemicals Health/Environm ental Toxicity

Definition. PBT chem ical substances possess characteristics of persistence (P) in the environm ent, accum u lation in biological organism s (bioaccum u lation (B)), and toxicity (T) that m ake them priority pollutants and potential risks to hum ans and ecosystem s. Prom inent exam ples of PBT chem ical substances include the insecticide, DDT and polychlorinated biphenyls (PCBs). This PBT category includes the boundary conditions, such as fish bioconcentration/bioaccum u lation factors and environm ental persistence values, that would determ ine inclusion in (or exclusion from ) the category, and standard hazard and fate tests to address P, B, and T concerns for the chem ical substances fitting the category description. For background inform ation and discussion of this PBT category, see http://www.epa.gov/oppt/newchem s/pbtpolcy.htm , and the proposed (October 5, 1998; 63 FR 53417) and final (Novem b er 4, 1999; 64 FR 60194) “C ategory for Persistent, Bioaccum ulative, and Toxic Chem ical Substances” published in the US Federal Register.
Haz a rd Concerns. Generally, PBT chem ical substances are chem ical substances that partition to water, sedim e nt or soil and are not rem oved at rates adequate to prevent their bioaccum ulation in aquatic or terrestrial species, with the potential to pose a risk via food chain toxicity. Boundaries. The following table summ arizes specific identification criteria and associated process for use in evaluating new chem ical PBT substances.
NEW CHEMICALS PROGRAM PBT CATEGORY CRITERIA AND PROCESS TSCA Section 5 Action 5(e) Order/Significant New Use Rule (SNUR) Exposure/release controls included in order; testing required Ban Pending Testing Deny comm e rcialization; testing results m ay justify rem oving chem ical from “high risk concern”
Persistence (transform ation half-life) >2 m onths >6 m onths

Bioaccum u lation* (Fish BCF or BAF) > 1000 > 5000 Toxicity Develop toxicity data where necessary, based upon various factors, including concerns for P, B, other physical/chem i cal factors, and toxicity.
* Chem icals m u st also m eet criteria for MW(<1000) and cross-sectional diam eter (<20 D , or <20 x 10 -8 cm ). BCF of 1000 and 5000 are equivalent to log Kow of 4.2 and 5, respectively.

98General Testing Strategy Tier 1 . Based upon SAR and professional judgme nt, the Agency identifies a newchemical substance as a possible PBT chemical substance.

Log Kow L iquid chrom a tography (OPPTS 830.7570/OECD 117) or generator column(OPPTS 830.7560) m e thod Ready biodegradability Ready biodegradability (OPPTS 835.3110/OECD 301) 6 m e thods (choose one, or an equivalent test): DOC DieAway, CO 2 Evolution, Modified MITI (I), Closed Bottle, Modified OECD Screening, Manom etric Respirom etry or Sealed-vessel CO 2 production test (OPPTS 835.3120) Hydrolysis in water (if, based upon SAR, susceptibility to hydrolysis is suspected) OPPTS 835.2110 (OECD 111)

Results If the m easured log Kow is <4.2 or if the test chem ical passes the ready biodegradability test (i.e., not persistent in the environm ent), no further PBT-related testing is required. If the m easured log Kow is greater than or equal to 4.2 and the chem ical does not pass the ready biodegradability test, the chem ical would require tier 2 testing. If hydrolysis testing is conducted and results in a half-life of <60 days, further testing m ay not be needed, but the need for testing m

u st be determ ined after consideration of factors specific to the case, such as physical/chem i cal properties, persistence and bioaccum u lative qualities of hydrolysis products, and the nature of the expected releases.

99Tier 2. Biodegradability and Bioaccumulation Biodegradability Shake-flask die-away test (OPPTS 835.3170) or an equivalent test. Bioaccum u lation F ish bioconcentration test (OPPTS 850.1730/OECD 305), or an equivalent test). Measured BCF should be based on 100 percent active ingredient and m easured concentration(s). Results If the m easured biodegradation half-life is >60 days and m easured BCF is >1000, tier 3 testing will be required. If only one condition is m e t, releases and exposure are further considered to determ ine if additional testing is required.
Tier 3 . Toxicity/advanced environmental fate testing.

Hum an health toxicity Com b ined repeated dose oral toxicity with the reproductive/developm ental toxicity screening test (OECD No. 422) in rats. Other health testing will be considered where appropriate. Environm ental fate Sedim e nt/water m i crocosm biodegradation test (OPPTS 835.3180). Chronic environm ental toxicity Fish (rainbow trout) and daphnids should be tested according to 40 CFR 797.1600 (sam e as OPPTS 850.1400/OECD 210) and 40 CFR 797.1330 (sam e as OPPTS 850.1300/OECD 202), respectively. Additional testing to evaluate other biota (e.g., avian, sedim e nt dwelling organism s) or other effects (e.g., endocrine disrupting potential) will be considered where appropriate.
Novem b er, 1999

100Category: Phenolphthaleins Hum an Health Definition. Any chem ical containing the phenolphthalein structure is considered to be a m e m b er of the category.

Haz a rd Concerns. The health concern for phenolphthalein and derivatives of phenolphthalein is for cancer based on the NTP cancer study (NTP report TR-465, Novem b er 1996) for phenolphthalein with adm i nistration via the di et. There was clear evidence of carcinogenic activity in m a le F344/N rats based on m arkedly increased incidences of benign pheochrom ocytom as of the adrenal m e dulla and of renal tubule adenom as and adenom as or carcinom as (com bined). There was som e evidence of carcinogenic activity in fem a le F344/N rats based on the increased incidences of benign pheochrom ocytom as of the adrenal m e dulla in the 12,000 ppm group and of benign or m a lignant pheochrom ocytom as (com bined) in the 12,000 and 25,000 ppm groups. There was clear evidence of carcinogenic activity in m a le B6C3F 1 mi c e based on increased incidences of histiocytic sarcom as and of m a lignant lym phom as of thym ic origin. There was clear evidence of carcinogenic activity in fem a le B6C3F 1 m i ce based on increased incidences of histiocytic sarcom as, m a lignant lym phom as of all types, lym phom as of thym ic origin, and benign sex-cord strom a l tum o rs of the ovary. Boundaries. No boundaries currently defined.

101Testing. EPA considers the following tests to be the m o st appropriate for phenolphthalein derivatives found to pose an unreasonable risk to hum an health: Tier 1: Com p arative derm al and oral absorption study in rats (OPPT Harm onized Test Guidelines, 870.7485) in vitro Chrom o som e aberrations study in Chinese ham s ter ovary cells with phenolphthalein as an additional positive control (OPPT Harm onized Test Guidelines, 870.5375) Log Kow test (OPPTS 830.7550)
W a ter solublility for both lactone and acid form (OECD 105) Ready biodegradability (OPPT 835.3110) Tier 2: 2-year carcinogenicity study in m i ce (OPPT Harm onized Test Guidelines, 870.4200) April, 1998

102Category: Phenols Environm ental Toxicity Definition. This category includes phenols (i.e., m onophenols), polyhydroxy phenols, and polyphenols. It is assum e d that these com pounds need to be absorbed to be toxic, therefore, com pounds with MW s > 1000 will be excluded from this category. Acute toxicity for phenols which are liquids at room tem p erature is known to be lim ited by the octanol/water partition coefficient (Kow ). Above a log K owvalue of about 7.38, phenols are expected to show no effects at saturation during 96-h exposures (Veith and Broderius 1987). Phenols which are solids at room tem p erature m ay show no toxicity at saturation at lower K owvalues depending on the m e lting point, i.e., the higher the m e lting point at a given K ow , the greater the likelihood that no toxicity will be observed at saturation. For solids, no effects at saturation has to be dem onstrated on a case-by-case basis. There are no known K owlim its f o r chronic toxicity at this tim e, but it m ay not be m u ch above a log K ow= 9.0 for liquid phenols. Future testing will determ ine this K ow lim it.
Haz a rd Concerns

. T he acute and chronic toxicity for phenols can be predicted through SAR Analysis. SARs are available for fish 96-h LC50, daphnid 48-h LC50, green algal 96-h EC50, fish chronic value (ChV), daphnid ChV, and al gal ChV. Mem b ers of this category exhibit toxicity ranging from low toxicity (i.e., > 100 m g /L) to high toxicity (i.e., < 1 m g /L) depending on their K ow , MW , and substitutions (e.g., dinitrophenols). Dinitrophenols are known to be m o re toxic than predicted by these SARs (see the category for polynitroarom a tics).
Fate : Phenols are subject to indirect photolysis under environm entally realistic conditions. Boundaries. There are no known lower boundaries. The upper boundaries will be based on Kow and MW

. Acute toxicity expected with log K ow< 7.38; no effects at saturation during 96-h exposures when log K ow>= 7.38. Chronic toxicity has no known upper bound for log K ow , but it is probably near 9. MW will be < 1000. The environm ental base set of tests will be requested f o r aquatic releases and the terrestrial base set of tests will be recomm e nded f o r terrestrial exposures. Wh en the log K owis >= 7.38, chronic toxicity testing with fish, daphnids, and green algae will be recomm e nded. General Testing Strategy
I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity tests for fish (40 CFR 797.1400) and daphnids (40 CFR 797.1300) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should equal the aqueous solubility lim it; and solvent can be

103used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. The algal toxicity testing (40 CFR 797.1050), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% AI and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; the highest treatm ent concentration on a nom inal-basis equal to the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. If there is no significant risk from the PMN after the results of the environm ental base set have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Direct and Indirect Photolysis Screening Test (40 CFR 796.3765). If t1⁄2 < 2 days, go to Tier 3; if t1⁄2 > 2 days, go to Tier 4. Tier 3a. If t1⁄2 < 2 days and photolysis products are known and/or identified, then assess photolysis products for environm ental hazards. Tier 3b. If t1⁄2 < 2 days and photolysis products are not known and/or identifiable, then prepare a stock solution of PMN using the standard hum ic-containing solution described in the direct and indirect photolysis screening te st [40.796.3765 (b)(2) and (c)(2)], expose to sunlight for at least 6 half-lives (t1⁄2), and test photolysis products for toxicity with m o st sensitive species f r om environm ental base set. For exam ple, the m o st sensitive species f r om the environm ental base set has an EC50 value = 2.0 m g PMN/L (based on 100% AI, therefore, prepare a 5.0 m g PMN per liter stock solution based on 100% AI using the standard hum ic-containing solution. This stock solution is exposed to sunlight for at least 6 half-lives to ensure that all of the PMN has been photolyzed, and then this stock solution is used to retest the m o st sensitive aquatic species to determ ine if the photolysis products of the PMN are m o re or less toxic that the PMN. Tier 4. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (40 CFR 797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% AI and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test because Van Leeuwen et al (1990) have dem onstrated that the 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen);

104Daphnid chronic toxicity testing ( 40 CFR 797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% AI and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; m e asured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d daphnid chronic toxicity test cannot be substituted f o r the 21-d toxicity test because Van Leeuwen et al (1990) have dem ons trated that the fish 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the fish 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen). Aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Aerobic Aquatic Biodegradation 40 CFR 796.3100 Modified Sturm Test 40 CFR 796.3260 Closed Bottle Test 40 CFR 796.3200 Modified OECD Screening Test 40 CFR 796.3240 Modified MITI Test (I) 40 CFR 796.3220 Modified AFNOR Test 40 CFR 796.3180
II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial exposures. Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay. References. Van Leeuwen CJ, Adem a DMM, and Herm ens J. 1990. Quantitative structure-activity relationships for fish early life stag e toxicity. Aquatic Toxicology 16:321-334. Veith GD and Broderius SJ. 1987. Structure-toxi city relationships for industrial chem icals causing type (II) narcosis syndrom e. In Kaiser KLE (ed), QSAR In Environm ental Toxicology - II, p 385-391. Reidel Publishing Com p any. July, 1991

105Category: Phosphates, Inorganic Definition. This category includes all inorganic soluble form s of phosphates, such as, phosphoric acid [PO4H3 or OP(=O)(O)O] and its salts or phosphate salts, pyrophosphates, polyphosphates, and organic and inorganic fo rm s of phosphorous that can be oxidized to phosphates rapidly. Inorganic form s of phosphonic acid (H2PO3 or OP(=O)O) are not included in this category because m onopotassium phosphonic acid [13977-65-6] has been shown not to be an algal nutrient, not to be a replacem ent for phosphate in algal growth m e dium , and not to cause exponential growth of green algae.
Haz a rd Concerns. The standard environm ental toxicity profile for inorganic phosphates as P in mgP / L i s : ECOTOX: Predicted (P) and m easured (M) toxicity values in m g /L (ppm ) are: fish 96-h LC50 > 100.0 P daphnid 48-h LC50 > 100.0 P green algal 96-h EC50 b < 0.030 P algal 96-h EC290 b = 0.030 M S,N fish chronic value > 10.0 P daphnid ChV > 10.0 P algal ChV = 0.010 M Hutchinson 1957 Predictions are based on SAR-nearest analog analysis for soluble form s of inorganic phosphates (PO4); SAR chem ical class = P-O4; MW of P = 31, PO4 = 95, PO4H3 = 98; pH7; effective concentrations based on 100% active ingredients and nom inal concentrations of P; hardness <24.0 m g /L as CaCO3; and TOC <2.0 m g /L; high concern for eutrophication; assessm ent factor = 10.0 concern concentration = 0.001 Phosphate phosphorus has the potential to stim ulate the growth of green algae and cause algal bloom s and eutrophication in freshwater and m arine environm ents. The phosphate anion is a plant nutrient and is the m a jor lim iting nutrient in m any freshwater environm ents. The USEPA OW W Q C (USEPA 1976) states: [W ]hen such concentrations [of total phosphate phosphorus]exceed 25 

### : g/l at the tim e of the spring turnover on a volum e-weighted basis in lakes or reservoirs, they m ay occasionally stim ulate excessive or nuisance growths of algae and other aquatic plants. Algal growths im part undesirable tastes and odors to water, interfere with water treatm e nt, becom e aesthetically unpleasant and alter the chem istry of the water supply. The contribute to the phenom enon of cultural eutrophication. To prevent the developm ent of biological nuisances and to control accelerated or cultural eutrophication, total phosphates as phosphorus (P) should not exceed 50 

### : g/l in any stream at the point where it enters any lake or reservoir, nor 25 

### : g/l within the lake or reservoir. A desired goal for the prevention of plant nuisances in stream s or other flowing waters not discharging
106directly to lakes or im poundm ents is 100 

### : g/l total P (Mackenthum , 1973). Most relatively uncontam inated lake districts are known to have surface waters that contain from 10 m i crogram s P/L (or 31 m i crogram s PO4H3/L) to 30 m i crog P/L (or 92 m i crog PO4H3/L) (Hutchinson, 1957). Odum (1971) called phosphorus a m a jor factor in the process of cultural eutrophication. Eutrophication results when nutrients, especially phosphates, are im ported into aquatic ecosystem s. This process is analogous to fertilizing an agricultural field. Phosphates act like a fertilizer and produce increases in the abundance of algae and other aquatic plants; eventually, they can result in excessive growths of algae (i.e., algal bloom s). Excessive algae can block sunlight from reaching subm erged m acrophytes and other m i croalgae below the surface. Wh en the algae die and fall to the bottom of slow-m oving water or lakes, they are consum ed by bacteria which use up available oxygen producing anaerobic conditions. Decaying algae cause odors and oxygen depletion of the water to occur with concom itant detrim ent to fish and aquatic invertebrates and, in turn, fishing and water-based recreational activities. Phosphate concentrations as low as 0.050 m g P/L (ppm ) or 50.0 m i crogram s P/L (ppb) will produce exponential growth of green algae in 96 hours (Miller et al. 1978), and phosphate concentrations from 10 to 60 m i crogram s P/L (ppb) were correlated to algal bloom s and oxygen depletion (i.e., eutrophication) in Lake W a shi ngton, Seattle (Odum 1971). Phosphates have been severely lim ited or banned from detergents in 13 states, the District of Colum b ia, and several counties and m unicipalities (USEPA 1992). Sodium tripolyphosphate was one of the principal com ponents of synthetic detergents. Freshwater green algae are clearly the m o st sensitive group of aquatic organism s to phosphate additions to water. Boundaries. The boundaries for inorganic phosphate com pounds depend on the release of inorganic phosphates which can be used by algae as a m i neral nutrient. Any inorganic phosphate which can be used by algae as a nutrient or which transform s to release or becom e s an algal nutrient is included in this category. Testing. Based on a consideration of available data on inorganic phosphates and the physicochem i cal properties of these com pounds, there is no need for further testing. However, if there is doubt about the availability of phosphate in a nutrient form from a PMN substance, then the algal toxicity test can be done with the PMN substance substituted for phosphate nutrient in the algal growth m e dium

. If a PMN substance is capable of stim ulating exponential growth green algae, it m u st be em phasized that inorganic phosphates should not be released to water because of their potential to cause eutrophication and their persistence. Regulatory Actions. (1) Phosphates have been severely lim ited or banned from detergents in 13 states, the District of Colum b ia, and several counties and m unicipalities (USEPA 1992). (2) W a ter Quality Criterion, 1976 (USEPA 1976): total phosphates as phosphorus (P) should not exceed 50 m i crogram s p/L in any stream at the point where it enters any lake or reservoir, nor 25 m i crogram s P/L within the lake or reservoir.

107(3) Hazard assessm ents of selected aqueous cleaner chem icals, USEPA (1990): OPPT recomm e ndation to the Office of Air and Radiation: "...phosphates should never be released to water." (4) Agency recomm e ndation for a "nationwide elim ination of the use of household laundry detergents containing phosphates." (USEPA 1992). References. Hutchinson, G. E., 1957, A treatise on limnology. New York, NY: John W iley & Sons. Mackenthun, K. M., 1973, Toward a cleaner aquatic environm ent. W a shington, DC: USEPA. Miller, W

. E., Greene, J. C., and Shiroyam aT. 1978. The Selenastrumcapricornutum Printz algal assay bottle test. Corvallis, OR: USEPA, Of f i ce of Research and Developm ental, Environm ental Research Laboratory-Corvallis, EPA-600-9-78-018. United States Environm ental Protection Agency (USEPA). 1976. Quality Criteria for W a ter. EPA-440/9-76-023. W a shington, DC: Office of W ater Planning and Standards, OW , USEPA. Available from NTIS, Springfield, VA 22161, PB-263-943. United States Environm ental Protection Agency (USEPA). 1990. (Health and environm ental effects of selected aqueous cleaner chem icals. W a shington, DC: Health and Environm ental Review Division, Office of Polluti on Prevention and Toxics, USEPA. United States Environm ental Protection Agency (USEPA). 1992. ( DRAFT Phosphate detergents: An evaluation of the benefits and co sts of elim inating their use in the United States. W a shington, DC: Environm ental Results Bran ch, Office of Policy, Planning, & Evaluation, USEPA. March, 2000

108Category: Phosphinate Esters Environm ental Toxicty Phosphinate esters are a class of organic com pounds characterized by the functional group R-O-P(=O)(R)R. Phosphinate esters are m e tabolically active and exhibit excess aquatic toxicity in addition to narcosis. Phosphinate esters exhibit excess toxicity relative to sim p le esters (-C(=O)O-) and phosphate esters (-OP(=O)(O-)O-. It is assum e d that phosphinate esters need to be absorbed to be toxic, therefore, phosphinate esters with MW >1000 will be excluded from this category. Acute toxicity for phosphinate esters is known to be correlated and lim ited by the octanol/water partition coefficient (Kow ). Above a log K owvalue of >5.0, phosphinate esters show no effects at saturation during 96-hr exposures to fish. Phosphinate esters which are solids at room tem p erature m ay show no toxicity at saturation at lower K owvalues depending on the m e lting point, i.e., the higher the m e lting point at a given K ow , the greater the likelihood that no toxicity will be observed at saturation. For solids, the no effects at saturation has to be determ ined on a case-by-case basis. There are no m easured upper K owlim its f o r chronic toxicity at this tim e, but it m ay not be m u ch above a log K ow= 8. Future testing will determ ine K owlim its.
Haz a rd Concerns. The toxicity for phosphinate esters has been determ ined through SAR analysis. SARs for freshwater species: log fish 96-h LC 50 (m illim oles/L) = -1.201 -0.260 log K ow where n=2, R2 =1.0, LOGKOW (SRC)<5, MW <1000; log daphnid 48-h LC 50 (m illim oles/L) = -1.501 -0.260 log K ow (SRC)<5, MW <1000; where n=2, R2 =1.0, LOGKOW
log green algal 96-h EC 50 (mm o l/L) = -1.974 -0.332 log K ow where n=2, R2 =1.0, LOGKOW (SRC)<6.4, MW <1000, based on test data for Pseudomonas putida ; log fish ChV (m illim oles/L) = -2.210 -0.499 log K ow where n=2, R2 =1.0, LOGKOW (SRC)<8, MW <1000, based on F96÷ACR10; log daphnid ChV (m illim oles/L) = -2.445 -0.407 log K ow where n=2, R2 =1.0, LOGKOW (SRC)<8, MW <1000, based on D48÷ACR10; and log algal ChV (mm o l/L) = -3.315 -0.223 log K ow where n=2, R2 =1.0, LOGKOW (SRC)<8, MW <1000, based on test data for Pseudomonas putida

. SARs for saltwater species: log fish 96-h LC 50 (m illim oles/L) = -1.160 -0.368 log K ow
109where n=2, R2 =1.0, LOGKOW (SRC)<5, MW <1000; and log my sid 96-h LC 50 (m illim oles/L) = -2,019 -0.476 log K ow where n=2, R2 =1.0, LOGKOW (SRC)<5, MW <1000.
Fate. Phosphinate esters hydrolyze in water and their rate of hydrolysis is correlated with pH: the m o re alkaline the faster the hydrolysis. The SAR for hydrolysis is: log hydrolysis t1⁄2 (d) = 4.325 -0.497 pH where n=2, R2 =1.0. At pH 7.1, the hydrolysis t1⁄2 = 6.3 days or 150 hours. Boundaries. MW <1000. Log K ow<5.0 for acute toxicity to fish and aquatic invertebrates; log K ow <6.4 for toxicity to green algae as a 96-h EC 50 ; and log K owassum e d to be <8.0 for chronic toxicity to aquatic organism s, but could be higher. General Testing Strategy. I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity test s for fish (OPPTS 850.1075) and daphnids (OPPTS 850.1010) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (ai) and m ean m easured concentrations; m easured TOC of dilution water in the control; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; and solvent can be used to assist the aldehyde to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the test chem ical significantly above its aqueous solubility lim it. The algal toxicity testing (OPPTS 850.5400), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (ai) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; and solvent can be used to assist the aldehyde to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the test chem ical significantly above its aqueous solubility lim it. If there is no significant risk from the PMN chem ical after the results of the environm ental base set have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2.

110Tier 2. Aerobic biodegradability according to either of the following test guidelines: Ready Biodegradability OPPTS 835.3110 Sealed Vessel CO2 Production Test OPPTS 835.3120 If there is no significant risk from the PMN chem ical after the results of biodegradation testing have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 3. Tier 3. Fish chronic toxicity testing, i.e., fish early life stage (ELS) toxicity testing (OPPTS 850.1400), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrati ons at days 7, 14, 21, and 28; m e asured TOC of dilution water in the control; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN significantly above its aqueous solubility lim it; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test because the 7-d ELS toxicity test m ay underestim ate chronic toxicity m easured by the 28-d ELS toxicity test when the Chronic Values are com p ared. Daphnid chronic toxicity testing (O PPTS 850.1300), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; m easured TOC of dilution water in the control; ideally, m ean m easured concentrations in the highest treatm e nt concentration should be equal the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN significantly above its aqueous solubility lim it; and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test because the daphnid 7-d short-term chronic toxicity test m ay underestim ate chronic toxicity m easured by the daphnid 21-d chronic toxicity test when the chronic values are com p ared.
II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test [O PPTS 850.4230], the earthworm toxicity test [OPPTS 850.6200], and the soil m i crobial comm unity bioassay [OPPTS 850.5100]) will be recomm e nded for terrestrial exposures. Chronic toxicity te sting for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay. September 1996.

111Category: Polyanionic Polymers (& Monomers) Environm ental Toxicity Definition. There are two subcategories of polym ers that are of concern: - polyarom a tic sulfonates - condensation products of sulfonated arom atics with form aldehyde, and - polyacrylates with free carboxyl groups. This category includes m onom ers with two or m o re acid groups and which act like organic acid chelators. The acid groups m ay include carboxylic acids, silicic acids, phosphoric acid, and sulfuric acids. These acids m ay also contain thiol substitutions. The acid groups on a m onom er m ay be all of the sam e type of acid or m ay be a m i xture of acids. The m onom eric nucleus m ay include carbon, silica, oxygen, sulfur, and nitrogen, or m i xtures of these elem ents. Mem b ers of this category m u st be water soluble or water self-dispersing. Molecular weights can be > 1000.
Haz a rd Concerns. Polyarom atic sulfonates are m oderately toxic to fish, daphnids, and algae. The polyacrylates are toxic to algae only. Th ere is no apparent relationship between charge density and toxicity, and no QSAR (quantitative structure-activity relationship) has been developed for these polym ers, or the polyanionic m onom ers. Polyanionic m onom ers (as salts, e.g., Na or K salts) are m oderately toxic to green algae, but show low toxicity to fish and daphnids. It is assum e d that the toxicity of these com pounds is due to over-chelation of nutrient elem ents needed f o r algal growth and that this toxicity will be m itigated in the presence of Ca ++ either added to the com pound before testing or present in the growth/test m e dium at a hardness of about 150 m g /L as CaCO 3 . Boundaries. Polym e rs m u st be water-soluble. Molecular weights are generally >1,000. General Testing Strategy To address ecotoxicity concerns, for polycar boxylic acids (polyacrylates) and polyanionic m onom ers, algal testing (static m e thods, nom inal concentrations) is recomm e nded, in 3 separate tests: 1) the test substance as is, 2) with equiva lent of calcium ion, and 3) with growth m e dium at 150 m g /L hardness, as CaCO 3 ). For polyarom a tic sulfonates, base set aquatic toxicity testing (flow through m e thods, m easured concentrations) in algae, daphnids, and fish is recomm e nded. Septem ber, 1988; revised April, 1991

112Category: Polycationic Polymers Environm ental Toxicity Definition. Any polym er that exists in the environm ent with m u ltiple positive charges is a m e m b er of this class. Such structures include polyam i nes, polyquaternary amm onium , polysulfonium , and polyphosphonium com pounds.
Haz a rd Concerns. Mem b ers of the category are toxic to fish, invertebrates, and algae. Algae are six-fold m o re sensitive than fish and daphnids. It is presum ed that these com pounds act on the surface of organism s and need not be absorbed. Toxicity increases exponentially with increasing charge density at cationic equivalent weights of >400. At lower charge equivalent weights, toxicity does not increase. A num ber of QSARs (quantitative structure-activity relationships) have been developed to predict the toxicity of polycationic polym ers. Boundaries. Polym e rs m u st be water-soluble or water-dispersible. Molecular weights are >300 although the typical new chem ical polycationic polym er has a m o lecular weight in excess of 1000. EPA has been engaged in discussions with the Cationic Flocculant Producers Association in an attem p t to identify and develop a set of tests on representative polycationic polym ers which could better define the lim its of the category. General Testing Strategy To address ecotoxicity concerns, for polym ers with % am ine nitrogen <0.7% and >0.1%: base set acute aquatic toxicity testing in algae, daphnids, and fish, plus hum ic acid testing in fish (20 m g /L hum ic acid in dilution water and 10 m g /L) is recomm e nded. All testing uses static m e thod, nom inal concentrations. For som e m e m b ers of the class, a test of sorbed chem ical using a benthic organism m ay also be recomm e nded. Septem ber, 1988; periodically revised.

113Category: Polynitroaromatics Environm ental Toxicity This category includes all dinitroarom a tics and trinitroarom a tic com pounds, for exam ple, dinitrobenzenes, dinitroanilines, dinitrophenols, and dinitropyridines. Polynitroarom a tics probably act by uncoupling of oxidative phosphorylati on (Doull et al 1980). It is assum e d that these com pounds need to be absorbed to be toxic, therefore, com pounds with MW s > 1000 will be excluded from this category. Acute toxicityfor polynitroarom a tics which are liquids at room tem p erature is assum e d to be lim ited by the octanol/water partition coef f i cient (K ow ). Above a log K owvalue of => 7.00 (based on test data for anilines reported Veith and Broderius 1987), polynitroarom a tics are not expected to show t oxic effects at saturation during 96-h exposures. Polynitroarom a tics which are solids at room tem p erature m ay show no toxicity at saturation at lower K owvalues depending on the m e lting point, i.e., the higher the m e lting point at a given K ow , the greater the likelihood that no toxicity will be observed at saturation. For solids, the no effects at saturation has to be determ ined on a case-by-case basis. There are no known K owlim its for chronic toxicity at this tim e, but it m ay not be m u ch above a log K ow= 10 for liquid polynitroarom a tics. Future testing will determ ine this K owlim it.
Haz a rd Concerns. The acute and chronic toxicity for several classes of polynitroarom a tics have been determ ined through SAR Analysis: dinitroanilines : fish 96-h LC50 (m M/L)= -0.027 -0.596 log K ow ; N=2; R2 =1.0 daphnid 48-h LC50 (m M/L)= -0.296 -0.558 log K ow

ow ; N=2; R2 =1.0 ; N=2; R2 =1.0 fish ChV (m M/L)= -0.91 -0.661 log K dinitrobenzenes : fish 96-h LC50 (m M/L)= -1.867 -0.333 log K ow ; N=2; R2 =1.0 daphnid 48-h LC50 (m M/L)= -0.325 -0.634 log K ow ; N=3; R2 =0.86 fish ChV (m M/L)= -3.0 -0.40 log K ow ; N=2; R2 =1.0 daphnid ChV (m M/L) = -0.7 -0.625 log K ow ; N=2; R2 =1.0 dinitrophenols : fish 96-h LC50 (m M/L)= -0.285 -0.559 log K ow ; N=4; R2 =0.96 daphnid 48-h LC50 (m M/L)= 0.083 -0.632 log K ow ; N=7; R2 =0.85 fish ChV (m M/L)= -1.78 -0.552 log K ow ; N=4; R2 =1.0 daphnid ChV (m M/L) = -0.465 -0.654 log K ow ; N=2; R2 =1.0 Polynitroarom a tics are known to be m o re toxic than predicted by the neutral organic SARs.
Mem b ers of this category exhibit toxicity rangi ng from low toxicity (i.e., > 100 m g /L) to high

owand MW

. toxicity (i.e., < 1 m g /L) depending on their K

114Fate : Polynitroarom a tics are expected to be subject to rapid direct and indirect photolysis under environm entally realistic conditions. Boundaries. : There are no known lower boundaries. The upper boundaries will be based on K owand MW

. Acute toxicity expected with log K ow< 7.0; no effects at saturation during 96-h exposures when log K ow>= 7.0. Chronic toxicity has no known upper bound for log K ow , but it is probably near 10. MW will be < 1000. The environm ental base set of tests will be requested f o r aquatic releases and the terrestrial base set of tests will be recomm e nded f o r terrestrial exposures. Wh en the log K owis >= 7.0, chronic toxicity testing with fish and daphnids will be recomm e nded. General Testing Strategy . I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity tests for fish (40 CFR 797.1400) and daphnids (40 CFR 797.1300) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should equal the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. The algal toxicity testing (40 CFR 797.1050), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; the highest treatm e nt concentration on a nom inal-basis equal to the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. If there is no significant risk from the PMN after the results of the environm ental base set have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Direct and Indirect Photolysis Screening Test (40 CFR 796.3765). If t1⁄2 < 2 days, go to Tier 3; if t1⁄2 > 2 days, go to Tier 4.

115Tier 3a. If t1⁄2 < 2 days and photolysis products are known and/or identified, then assess photolysis products for environm ental hazards. Tier 3b. If t1⁄2 < 2 days and photolysis products are not known and/or identifiable, then prepare a stock solution of PMN using the standard hum ic-containing solution described in the direct and indirect photolysis screening te st [40.796.3765 (b)(2) and (c)(2)], expose to sunlight for at least 6 half-lives (t1⁄2), and test photolysis products for toxicity with m o st sensitive species f r om environm ental base set. For exam ple, if the m o st sensitive species f r om the environm ental base set has an EC50 value = 2.0 m g PMN/L (based on 100% active ingredients [AI]), then prepare a 5.0 m g PMN per liter stock solution based on 100% AI using the standard hum iccontaining solution. This stock solution is exposed to sunlight for at least 6 half-lives to ensure that all of the PMN has been photolyzed, and then this stock solution is used to retest the m o st sensitive aquatic species to determ ine if the photolysis products of the PMN are m o re or less toxic that the PMN. Tier 4. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (40 CFR 797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test because Van Leeuwen et al ( 1990) have dem onstrated that the 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen); Daphnid chronic toxicity testing ( 40 CFR 797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; m easured TOC of dilution water in the control; the highest treatm ent concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test because Van Leeuwen et al (1990) have dem onstrated that the fish 7-d ELS toxicity test underestim ated the chronic toxicity of anilines

116m easured by the fish 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen). Aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Aerobic Aquatic Biodegradation 40 CFR 796.3100 Modified Sturm Test 40 CFR 796.3260 Closed Bottle Test 40 CFR 796.3200 Modified OECD Screening Test 40 CFR 796.3240 Modified MITI Test (I) 40 CFR 796.3220 Modified AFNOR Test 40 CFR 796.3180
II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial exposur es. Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay. References. Doull J, Klaassen CD, and Am dur MO. 1980. Casarett and Doull' s Toxicology. The Basic Science of Poisons. Second Ed. New York, NY: Macm illan Pub. Co., Inc. Van Leeuwen CJ, Adem a DMM, and Herm ens J. 1990. Quantitative structure-activity relationships for fish early life stag e toxicity. Aquatic Toxicology 16:321-334. Veith GD and Broderius SJ. 1987. Structure-toxi city relationships for industrial chem icals causing type (II) narcosis syndrom e. In Kaiser KLE (ed), QSAR In Environm ental Toxicology - II, p 385-391. Boston, MA: D. Reidel Publishing Com p any. May, 1991

117Category: Respirable, Poorly Soluble Particulates Health Only Definition. This category includes a variety of inor ganic, poorly soluble (as designated in ILSI 2000) particulates. Typically, they are oxides of various m e tals or nonm etals (i.e., silicon) Boundaries. Potential f o r respirability--that is, if there are any particles 

### # 10 

### : in diam eter in the m a terial being handled by workers. Summ arized below are currently available test data on five different poorly soluble particulates–s ilica, talc, titanium dioxide, PMN 96-175 (lithium m anganese oxide), and carbon black. The suitability of one or m o re of these analogues for a particular PMN particulate m u st be determ ined on a case-by-case basis. Risk is to be assessed by the m argin of exposure m e thod for the reason stated in the next paragraph.
Haz a rd Concerns The category concerns discussed here are lim ited to effects on the lung as a result of inhaling the particles. Broadly, as shown in rat inhalation studies, these effects range from inflamm a tion to fibrosis to, potentially, cancer. Because it is still not known with certainty whether high lung burdens of poorly soluble particulates can lead to lung cancer in hum ans via m e chanism s sim ilar to those of the rat, in the absence of m echanistic data to the contrary, it m u st be assum e d that the rat m odel can identify potential carcinogenic hazards to hum ans. Since the apparent responsiveness of the rat m odel at overload is dependent on coexistent chronic active inflamm a tion and cell proliferation, at lower lung doses in which chronic active inflamm a tion and cell proliferation are not present, no lung cancer hazard is anticipated (ILSI 2000). Som e of the particulates m ay contain m e tals, for exam ple, chrom i um , that m ay present other and m o re imm i nent toxicities, depending on the bioavailability of the m e tal ions. Thus, the toxicities of the m e tal com ponents of the particulates m u st also be assessed, and on a case-by-case basis. Lung Toxicity of Five Poorly Soluble Particulates 1. Fibrogenicity/Carcinogenicity of Crystalline Silica
Hum ans . Based on “sufficient evidence” for the carcinogenicity of inhaled crystalline silica from occupational sources, the International Agency for Research on Cancer (IARC, 1997) has classified crystalline silica as a Group 1 carcinogen in hum ans. According to IARC, studies of the following populations provided the least-confounde d exam inations of an association between silica exposure and cancer risk: gold m i ners, stone industry workers, granite shed and quarry workers, crushed stone industry workers, diatom aceous earth industry workers, refractory brick workers, pottery workers, and cohorts of registered silicotics. Not all of these studies dem onstrated excess cancer risks; but, in view of the large num ber of epidem iological studies undertaken and the wide range of populations and exposure circum stances, som e nonuniform ity of results would be expected. In som e studies, increasing risk gradients have been observed in relation to dose surrogates--cum ulative exposure, duration of exposure, or the presence of radiographically defined silicosis--and, in one instance, to peak intensity exposure. For these reasons, the IARC W o rking Group

118concluded that overall, the epidem iological findings support increased lung cancer risks from inhaled crystalline silica resulting from occupational exposure. Anim als

. IARC (1997) has concluded that there is “sufficient evidence” in experim e ntal anim als for the carcinogenicity of crystalline silica. A num ber of studies ( rev.in : IARC, 1997; W oo et al. , 1988; Holland, 1990) have dem onstrated that, in addition to silicosis/fibrosis, pulm onary tum o rs are induced in rats exposed to quartz (Mini-U-Sil; a comm on form of crystalline silica) by inhalation, single intrapleural, intraperitoneal, and/or intratracheal adm i nistration. In a 24- m onth inhalation study in rats, the lowest tested dose of respirable crystalline silica particles to induce lung tum o rs (Low est Observed Adverse Effect Level, or LOAEL) w a s 0.74 mg/m 3

. 

### K Note for Cases Using Silica as Primary Analogue: For LOAEL doses, the Agency considers a Margin of Exposure (MOE) of 1,000 or greater as representing a low order of toxicity for health endpoints for which a threshold dose is deem ed appropriate. In this instance, exposures would have to be reduced to 0.0007 m g /m 3 to achieve a MOE of 1,000, a concentration that is practically im possible to achieve. Because the LOAEL of 0.74 m g /m 3 is based on test data on pure silica and because silica is considered to be a m u ch m o re potent lung toxicant than crystalline particulates containing silica, the Agency has adopted the NIOSH REL (Recommended Exposure Level) for silica, 0.05 mg/m 3 , as a NCEL1/ (NewChemical Exposure Li m i t- see ht t p : //www.ep a.gov/ oppt / n ewchem s/ ncel m a i n .ht m f o r m o re information) concentration that w o uld adequately protect w o rkers from crystalline particulates containing silica. This NCEL replaces the former one based on OSHA Permissable Exposure Limit (PEL) for silica 0.1 mg/m 3

.

2. Fibrogenicity/Carcinogenicity of Talc
Hum ans . Epidem iology studies suggest an association between nonfibrous talc (a finely powdered hydrous m a gnesium silicate) and lung cancer risk. The OSHA PEL for talc is 20 m

ppcf (m illion particles per cubic f oot), and the NIOSH-recomm e nded 8-hr TWA concentration is 2 m g /m 3

. Anim als

. In an NTP inhalation study (NTP TR 421, 1993), m a le and fem a le F344 rats were exposed to 0, 6, or 18 m g /m 3 talc for 113 and 122 weeks, respectively. There was clear evidence of the carcinogenicity of talc in fem a le rats based on the increased incidence of alveolar/bronchiolar adenom as and carcinom as of the lung in the 18-mg/m 3 group (LOAEL). The No Observed Adverse Effect Level (NOAEL) from this study is 6 mg/m 3

. The NCEL for analogous particulates would be 0.06 m g /m 3

.

3. Fibrogenicity/Carcinogenicity of Titanium Dioxide (TI)
Hum ans . According to IARC (ACGIH 1993), there is inadequate evidence for th e carcinogenicity of TI in hum ans. [There is lim ited evidence for the carcinogenicity of TI in experim e ntal anim als.] Overall evaluation: TI is not classifiable as to its carcinogenicity to

119hum ans. The OSHA PEL’s for nuisance dust (in which TI is included) are 15 m g /m 3 total particulates and 5 m g /m 3 respirable fraction. The ACGIH TLV-TWAis 10 m g /m 3 of total dust containing no asbestos and <1% crystalline silica. The various literature case reports of lung toxicity in TI-exposed workers are generally inconclusive because of several factors--sm oking; other conditions, such as tuberculosis; or incom p lete data on TI exposure histories. In one fairly com p rehensive cohort study, Chen and Fayerweather (1988, as summ arized in ACGIH 1993) analyzed lung cancer m o rtality and inci dence of chronic respiratory disease, chest x-ray abnorm a lity, and pulm onary fibrosis am ong 1,576 workers exposed to TI for at least 1 yr at two production facilities between 1935 and 1984. Of the 211 deaths, 14 were attributed to respiratory cancer and 11 to nonm alignant respiratory disease com p ared with 18.3 and 13.8 cases expected, respectively, based on US m o rtality data. Expected deaths based on com p any m o rtality data since 1957 were 16.6 from respiratory cancer and 8.3 from nonm alignant respiratory disease. The authors concluded that the increase in deaths from nonm alignant respiratory disease com p ared with com p any data was not statistically significant. They further concluded that there was no significant associati on between exposure to TI and the respiratory diseases studied. Anim als

. There is lim ited evidence for the carcinogenicity of TI in experim e ntal anim als. Twoyear inhalation studies in anim als are summ arized below. (1) Lee et al. (1986, as cited in ACGIH 1993 and in USEPA 1983) exposed Charles River-CD rats to 0, 10, 50, or 250 m g /m 3 for 6 hr/day 5 days/wk for 2 yr. The m ean respirable fractions were 93.7% or greater. Squam ous cell carcinom as developed after exposure to the highest dose for the full 2 yr. The authors considered the carcinom as to be unique, experim e ntally induced tum o rs in rats and to be of questionable relevance to hum ans. Given the am ount of dust in the rat lungs, the tum o rs were postulated to be the result of saturation of norm a l pulm onary clearance m echanism s . There were m i nute areas of lung fibrosis at the two higher exposure levels.

At 50 m g /m 3 , m a ssive accum u lations of dust-laden m acrophages, foamy dust cells, and free particles were considered indicative of such an overload. At 10 m g /m 3 , the architecture of the air spaces rem a ined intact and there was no significant form ation of scar tissue. The NOAEL is 10 mg/m 3

. (2) Heinrich et al. (1995, abstract only) exposedW i star rats to diesel engine exhaust, carbon black, or ultrafine TI for 2 yr. Particle exposur e concentrations were increased during the course of the experim e nt. The average particle concentration of TI was 10 m g /m 3

. Lung tum o r rates (not given) increased with increasing cum u lative particle exposure tim es, independent of the type of particle. (3) Trochim o wicz et al. (1988, abstract only) exposed Charles River-CD rats to 10, 50, or 250

120mg /m 3 TI for 6 hr/day, 5 days/wk for 104 wk. Microscopic findings included alveolar cell hyperplasia, dust deposition, and m i nute areas of collagen fiber deposition associated with cholesterol giant cell granulom as. The earliest indicator of TI exposure was alveolar cell hyperplasia. Lung tum o rs were only found in rats exposed to 250 m g /m 3

. The NOAEL from this study is 10 mg/m 3

. On the basis of the NOAEL’s from the above studies, the NCEL for analogous particulates would be 0.1 m g /m 3

. However, this would m ake the NCEL for TI analogues identical to that for silica, a highly potent lung toxicant that IARC (1997) has classified as a Group 1 carcinogen in hum ans (see section 1, above). Therefore, it was decided to use the OSHA PEL for TI, which is 5 m g /m 3 (respirable fraction), as the NCEL for TI analogues. 4. Lung Toxicity of PMN 96-175, Lithium Manganese Oxide Anim al Data Only

. Rats were exposed by inhalation to 0 (air controls), 1, 4, or 16 m g /m 3 of the PMN substance 6 hr/day, 5 days/wk for 90 days. At the highest concentration, rats developed inflamm a tion of the lung and larynx. Sim ilar results were seen in the 14-day range-finding study at concentrations ranging from 10 to 100 m g /m 3

. The NOAEL from this study is 4 mg/m 3

. The NCEL for analogous particulates would be 0.04 m g /m 3

.

5. Fibrogenicity/Carcinogenicity of Carbon Black Chronic inhalation of carbon black produced pulm onary inflamm a tion, fibrosis, and lung tum o rs in rats (Nikula et al. 1995, Heinrich et al . 1995). These effects were observed at dust concentrations of 2.5, 6.6, and approxim a tely 10 m g /m 3 carbon black. The LOAEL from these studies is 2.5 mg/m 3

. The NCEL for analogous particulates would be 0.0025 m g /m 3 . [The OSHA PEL for carbon black is 3.5 m g /m 3 .]
General Testing Strategy A 90-day inhalation toxicity test (CFR 870.3465) in rats with special attention to histopathology (inflamm a tion and cell proliferation) of the lung tissues and to various param e ters of the bronchoalveolar lavage fluid (BALF), e.g., m arker enzym e activities, total protein content, total cell count, cell differential, and cell viability. It is not necessary to look at internal organs. It is recomm e nded that a recovery period of 60 days be included to assess the progression or regression of any lesions. If the results of th e subchronic study indicate that the PMN particles have carcinogenic potential, a 2-year inhalation bioassay in rats m ay be warranted.

REFERENCES ACGIH. 1993. Am erican Conference of Governm e ntal Industrial Hygienists: Threshold Lim i t Values for Chem ical Substances and Physi cal Agents and Biological Exposure Indices, 1993-

1211994. Cincinnati, Ohio: ACGIH. Heinrich U, Fuhst R, Rittinghausen S, Creutzenberg O, Bellm ann B et al. 1995. Chronic inhalation exposure of W i star rats and two differe nt strains of m i ce to diesel engine exhaust, carbon black, and titanium dioxide. I nhal Toxicol 7:533-556. Abstract only. Holland LM. Reg. Toxicol. Pharm acol. 12 :224-237, 1990 IARC. 1997. International Agency for Research on Cancer. Monographs. Vol. 68, pp.41-336. ILSI. 2000. International Life Sciences Institute. W o rkshop: The relevance of the rat lung response to particle overload for hum an risk assessm ent. Inhal Toxicol 12: 1-17. Nikula KJ, Snipes MB, Griffith W C , Henderson RF, Mauderly JL. 1995. Com p arative pulm onary toxicities and carcinogenicities of chronically inhaled diesel exhaust and carbon black in F344 rats. Fund Appl Toxicol 25:80-94. NTP. Toxicology and Carcinogenesis Studies of Ta lc in F334/N rats and B6C3F1 m i ce. National Toxicology Program , Technical Report No. 421, 1993. Trochim o wicz HJ et al. 1988. J A ppl Toxicol 8:383-435. Abstract only. USEPA. 1983. Status report for 8EHQ-1083-0497. Prelim inary findings from a 2-yr rat inhalation study of titanium dioxide conducted by E. I. Du Pont de Nem ours & Co., Inc. From Justine W e lch, Chief, Chem ical Risk Indentifi cation Section, to Frank Kover, Chief, Chem ical Screening Branch, Existing Chem icals Assessm ent Div., OPPT, USEPA.
W oo, Y.-T., Lai, D.Y., Arcos, J.C., and Argus, M.F.: Chem ical Induction of Cancer, Vol. III C, Academ ic Press, p. 508, 1988. October, 2002

122Category: Rosin Environm ental Toxicity This category includes rosin, abietic acid, abietinic acid, sylvic acid, their salts, and polym eric form s whose MW < 1000. The m ode of toxic action of rosin is unknown. It is assum e d that these com pounds need to be absorbed to be toxic, therefore, com pounds with MW s > 1000 will be excluded from this category. It is also assum e d that these com pounds need to be water soluble to be toxic, therefore, com pounds with water solubilities (W S) < 0.001 m g /L (ppm ) will also be excluded f r om this category.
Haz a rd Concerns. The acute toxicity for rosin towards fish has been m easured to be between 0.410 to 0.700 m g /L by Leach and Thakore (1976 and 1978). Toxicity to aquatic invertebrates and green algae have no been reported. Based on these data, rosin presents a high concern for toxicity towards the aquatic environm ent. Boundaries. The only lower boundary is water solubility which is set at 0.001 m g /L (ppm ). The only upper boundary will be set at a MW > 1000 for stable com pounds. The environm ental base set of tests will be requested f o r aquatic releases and the terrestrial base set of tests will be recomm e nded for terrestrial exposures. General Testing Strategy . The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity tests for fish (CFR §797.1400) and daphnids (CFR §797.1300) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should equal the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility. The algal toxicity testing (CFR §797.1050), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; the highest treatm e nt concentration on a nom inal-basis equal to the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artif icially enhance the water solubility. Fish chronic toxicity testing, i.e., fish early life stage (ELS) toxicity testing (CFR §797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test because Van Leeuwen et al

123(1990) have dem onstrated that the 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen); Daphnid chronic toxicity testing (C FR §797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; m easured TOC of dilution water in the control; the highest treatm ent concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility; and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test because Van Leeuwen et al (1990) have dem onstrated that the fish 7-d ELS toxicity test underestim ated the chronic toxicity of anilines m easured by the fish 28-d ELS toxicity test by >5.3 tim es when the NOECs were com p ared (see Table VII in Van Leeuwen). The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test [§797.2800], the earthworm acute toxicity test [§795.150], and the soil m i crobial comm unity bioassay [795.3700]) will be recomm e nded for terrestrial exposures. Chronic toxicity testing for terrestrial organism s include: the plant whol e life cycle test [§797.2830], the plant uptake test [§797.2850], and the soil m i crobial comm unity bioassay. References. Leach , J. M. and Thakore, A. N., 1976. Toxic constituent in m echanical pulping effluents. Tappi, 59:129. Leach , J. M. and Thakore, A. N., 1978. Com pounds toxic to fish in pulp m ill waste stream s. Progr. W a ter Technol. (G.B.) 9:787. Van Leeuwen CJ, Adem a DMM, and Herm ens J. 1990. Quantitative structure-activity relationships for fish early life stag e toxicity. Aquatic Toxicology 16:321-334. January, 1994

124Category: Stilbene, derivatives of 4,4-bis(triaz i n-2-ylamino)
- Hum an Health (case-by-case) Definition. Any water soluble (sulfonated) derivativ e of 4,4-bis(triazin-2-ylam ino) stilbene is considered to be a m e m b er of this category.

Haz a rd Concerns. EPA previously identified hazard concerns for the possible developm ental/reproductive toxicity of new chem icals falling into this stilbene, derivatives of 4,4-bis(triazin-2-ylam ino)- category based on published test results for a m e m b er of the category. EPA has reviewed test data developed by the Stilbene Wh itening Agent Task Force of the Ecological and Toxicological Association of Dyes and Organic Pigm ents Manufacturers (ETAD). These studies consisted of a range-f inding and def i nitive 2-generation rat study (gavage) and range-f inding and def i nitive developm ental toxicity studies in rats and rabbits (gavage) for C.I. Fluorescent Brightener 220 (CAS No. 16470-24-9). Based on the results from these studies, EPA has concluded that available inform ation does not support continued identif ication of stilbenes as a new chem ical category presenting concerns f o r possible developm ental/reproductive toxicity. EPA will continue to evaluate the potential health concerns f o r new chem ical stilbenes that are notif ied but, with this change, will not apply a category understanding as regards potential developm ental/reproductive toxicity. References. Breslin, W J

. A pilot prenatal developm ental t oxicity study of C.I. Fluorescent Brightener 220 and C.I. Fluorescent Brightener 339 adm i nistered via oral gavage to rats. MPI Research Laboratory Study Identification 795-001, August 18, 1998a. Breslin, W J

. A pilot prenatal developm ental t oxicity study of C.I. Fluorescent Brightener 220 and C.I. Fluorescent Brightener 339 adm i nistered via oral gavage to New Zealand Wh ite rabbits. MPI Research Laboratory Study Identification 795-002, August 18, 1998b. Turck, PA. Prenatal developm ental toxic ity study of C.I. Fluorescent Brightener 220 adm i nistered via oral gavage to rats. MP I Research Laboratory Study Identification 795-003, Decem ber 2, 1999.

125Turck, PA. Range-finding reproduction study of C.I. Fluorescent Brightener 220 adm i nistered via oral gavage to rats. MPI Research Laboratory Study Identification 795-005, June 8, 2000a. Turck, PA. Prenatal developm ental toxic ity study of C.I. Fluorescent Brightener 220 adm i nistered via oral gavage to New Zealand Wh ite rabbits. MPI Research Laboratory Study Identification 795-004, January 27, 2000b. Turck, PA. Two generation reproduction and fertility study of C.I. Fluorescent Brightener 220 adm i nistered via oral gavage in rats. MP I Research Laboratory Study Identification No. 795- 006, August 24, 2001. Previous Category Statement References. Burg, AW , Rohovsky, MW & Kensler, CJ (1977). Current status of hum an safety and environm ental aspects of fluorescent whitening agents used in detergents in the United States. Critical Review & Environm ental Control, 7, 91-120. Gloxhuber, CH & Bloching, H (1978). Toxicologi c properties of fluorescent whitening agents. Clinical Toxicology, 13, (2), 171-203. Keplinger, ML, Fancher, OE, Lym an, FL & Calandra, JC (1974). Toxicologic studies of four fluorescent whitening agents. Toxicology & Applied Pharm acology, 27, 494-506. Keplinger, ML, Lym an, FL & Calandra, JC ( 1975). Three-generation reproduction studies with FWAs. In: Flourescent Wh itening Agents. R. Anliker & G. Muller (Eds.). Stuttgart, Germ any: Georg Thiem e Publishers.
Kram er, JB (1992). Fluorescent whitening agents. In: The Handbook of Environm ental Chem istry, 3, (part F), 351-366. O. Hutzinger (Ed.). Heidelberg, Germ any: Springer-Verlag Berlin. Lorke, D (1975). Studies of em bryo toxicity in rats & rabbits. In: Flourescent Wh itening Agents. R. Anliker & G. Muller (Eds.). Stuttgart, Germ any: Georg Thiem e Publishers. Lym an, FL, Schulze, J, Ganz, CR, Stensby, PS, Keplinger, ML & Calandra, JC (1975). Longterm toxicity of four fluorescent whitening agents. Food, Cosm etics & Toxicology, 13, 521-527. Poiger, T & Giger, W (1991). Determ ination of fluorescent whitening agents in sewage and sewage sludge by high perform ance liquid chrom a tography. Reprint of a poster presented at the m eeting of the Swiss Chem ical Society in Bern, Switzerland. October 18, 1991. EAWAG/ETH, CH-8600 Dubendorf, Switzerland. Zinkernagel, R (1975). Fluorescent whitening agents in the environm ent. In: Flourescent Wh itening Agents. R. Anliker & G. Muller (Eds.). Stuttgart, Germ any: Georg Thiem e Publishers. 8E-CAP - 0024. Report to Ciba-Geigy Corporati on. Teratogenic study with FA-15 in Albino

126rabbits, May 23, 1972. Subm itted to USEPA on October 9, 1992. PB81-148819. Inform ation Hazard Profiles on Potential Occupational Hazards. Vol. 2. Chem ical Classes Fluorescent Wh itening Agent (FWA ' s ). National Institute f o r Occupational Safety and Hazard, Rockville, Maryland. Decem ber 1979. January, 1992; revised Decem ber, 1994; revised August, 2002.

127Category: Thiols Environm ental Toxicity This category includes all thiols or m e rcaptans. It is assum e d that thiols have to be absorbed to be toxic, therefore, com pounds with MW s > 1000 will be excluded from this category. Acute toxicity for thiols which are liquids at room tem p erature is known to be lim ited by the octanol/water partition coef f i cient (Kow ). Above a log K owvalue of <6.5 (CLOGP), thiols show no effects at saturation during 96-h exposures. Th iols which are solids at room tem p erature m ay show no toxicity at saturation at lower K owvalues depending on the m e lting point, i.e., the higher the m e lting point at a given K ow , the greater the likelihood that no toxicity will be observed at saturation. For solids, the no effects at saturation has to be determ ined on a case-by-case basis. There are no known K owlim its for chronic toxicity at this tim e, but it m ay not be m u ch above a log K owof 8 to 9 f o r liquid thiols. Future testing will determ ine this K owlim it.
Haz a rd Concerns. The acute toxicity for thiols has been determ ined through SAR Analysis: fish 96-h LC50 (m illim oles/L) = -1.022 - 0.447 log K ow daphnid 48-h LC50 (m illim oles/L) = -3.2 - 0.097 log K ow Effective concentrations for green algal 96-h EC50 should be assum e d to be sim ilar to fish and daphnids. Thiols which are substituted with a carboxylic acid will be about 10 tim es less toxic than predicted by this SAR at pH 7. Use the K owfor the free acid.
Mem b ers of this category exhibit toxicity rangi ng from low toxicity (i.e., > 100 m g /L) to high

ow , MW and substitutions. toxicity (i.e., < 1 m g /L) depending on their K Boundaries. There are no known lower boundaries. The upper boundaries will be based on Kow and MW

. Acute toxicity expected with log K ow<6.5; no effects at saturation during 96-h exposures when log K ow> 6.5. Chronic toxicity has no known upper bound for log K ow , but it is probably near 9. MW will be <1000. The environm ental base set of tests will be requested for aquatic releases and the terrestrial base set of tests will be recomm e nded f o r terrestrial exposures. Wh en the log K owis > 6.5, chronic toxicity testing will be recomm e nded. General Testing Strategy

I. Release to Aquatic Ecosystem s : Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity tests for fish (CFR §797.1400) and daphnids (CFR §797.1300) will be done using the flow-through m e thod with m easured concentrations; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should equal the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it.

128The algal toxicity testing (CFR §797.1050), should be done with static m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with at least 0.300 m g /L EDTA as a final concentration; the highest treatm e nt concentration on a nom inal-basis equal to the aqueous solubility lim it; and solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it. If there is no significant risk from the PMN after the results of the environm ental base set have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Aerobic Aquatic Biodegradation 40 CFR 796.3100 Modified Sturm Test 40 CFR 796.3260 Closed Bottle Test 40 CFR 796.3200 Modified OECD Screening Test 40 CFR 796.3240 Modified MITI Test (I) 40 CFR 796.3220 Modified AFNOR Test 40 CFR 796.3180 If there is no significant risk from the PMN after the results of the aerobic biodegradation have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 3. Tier 3. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (CFR §797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; m easured TOC of dilution water in the control; the highest treatm e nt concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d ELS stage toxicity test cannot be substituted for the 28-d ELS toxicity test. Daphnid chronic toxicity testing (C FR §797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; m easured TOC of dilution water in the control; the highest treatm ent concentration on a nom inal-basis should be set at the aqueous solubility lim it; solvent can be used to assist the PMN to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the PMN beyond its aqueous solubility lim it; and the 7-d daphnid chronic toxicity test cannot be substituted for the 21-d toxicity test.
II. Release to Terrestrial Ecosystem s

129The terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded f o r terrestrial exposures. Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay.
Decem ber, 1994

130Category: Substituted Triaz i nes Environm ental Toxicity This category includes substituted triazines which can be arom atic, partially arom atic (or partially saturated) and unsaturated. The nitrogens in the triazine ring m ay be symm e trical or asymm e trical. Substitutions on the carbons m ay include but not be lim ited to: aliphatic alcohols; ketones; benzene and substituted benzenes; aliphatic hydrocarbons, alkyenes and alkynes; free am ines and substituted am ines; cyclic aliphatic hydrocarbons; halogens; am ides; cyanides; ethers; m e thoxy groups; sulfides; azido groups; and carboxylic acid esters. Substitutions on the nitrogens m ay include but not be lim ited to: f r ee am ines and substituted am ines; -N=CH; aliphatic hydrocarbons, alkyenes and alkynes; and benzene and substituted benzenes.
Haz a rd Concerns. Many m e m b ers of this category are comm e rcial herbicides which are used to control both aquatic plants and terrestrial plants. Their m ode of toxic action is generally considered to be inhibition of photosynthesis. Ma ny m e m b ers of this class are toxic to algae at < 1 m g /L and toxic to terrestrial vascular plants at < 1 m g /kg. Mem b ers of this group can also be highly toxic to fish and aquatic invertebrates. Toxicity is expected to be related to the octanol/water partition coef f i cient with respect to f i sh and aquatic invertebrates, but toxicity to plants m ay not be related to Kow when log Kow < 5. Wh en the log Kow is < 5, algae and terrestrial plants are expected to be the m o st sensitive species. As log Kow increases, species differences are expected to dim i nish. At this tim e there is no form alized SAR for this category f o r any species. Toxicity predictions will be m a de using either the closest analog or averaging data for the two closest analogs which bracket the chem ical under question.

Boundaries. There are no known lower boundaries. The upper boundaries are based on Kow and MW

. Wh en the log Kow value is < 5 m g /L, the environm ental base set of tests will be requested f o r aquatic releases and the terrestrial base set of tests will be recomm e nded f o r terrestrial exposures. Wh en the log Kow is between 5 and 8, only chronic toxicity testing will be recomm e nded. Wh en the log Kow is > 8, no testing will be requested because no toxic effects at saturation will be expected. Generally, m e m b ers of this category will have MW s of less than 1000. General Testing Strategy The aquatic base set of environm ental toxicity tests will be recomm e nded for aquatic exposures and the terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm acute toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded for terrestrial exposures. August, 1989

131Category: Triarylmethane Pigments/Dyes w i th Non-solubiliz ing Groups Hum an Health Environm ental Toxicity Other Names: Triphenylm e thane Pigm ents/Dyes, Diphenylnaphthylm e thane Pigm ents/Dyes Definition. Structurally, triarylm ethane pigm ents/dyes are derivatives of triphenylm e thane or diphenylnaphthylm e thane. In order to achieve the required spectral absorption properties that characterize these dyes, am ine groups (prim ary, secondary, or tertiary) or hydroxyl groups m u st be present on the arom atic ring positions para to the m e thane carbon. Am ine substitutions are m o re prevalent than hydroxy substitutions (C.I. 42000-42175 for diam ino derivatives, C.I. 42500-42800 for triam i no derivatives, C.I. 43800-43875 for hydroxy derivatives, and 43500- 43570 for am inohydroxy derivatives).

Haz a rd Concerns. There are oncogenicity concerns for the triarylm ethane pigm ents/dyes based on analogy to Gentian Violet and C.I. Basic Red 9. In addition, there are developm ental and reproductive toxicity concerns for these com pounds based on analogy to Gentian Violet and Malachite Green. Ecotoxicity concerns are base d on QSAR predictions for delocalized cationic

132dyes. Cationic dyes are an established ecotoxicity category. Boundaries. Pigm ents/Dyes included in this category are the di- and tri-am ino substituted triphenylm e thane and diphenylnaphthyl-m ethane derivatives. Dyes substituted with solubilizing groups such as carboxylic acid, sulfonic acid, or ha logens, are not included. Pigm ents that have essentially negligible water solubility (<1ppb) and therefore, little or no bioavailability, are also not included. General Testing Strategy The New Chem icals Program considers the following tests to be appropriate to address the potential for triarylm ethane pigm ents/dyes to pose a significant risk. For Health - 1. Developm ental toxicity study in two species of rodents by the oral route (40 CFR 798.4900). 2. Two-generation reproductive toxicity study in rodents by the oral route (40 CFR 798.4700). 3. Salm onellatyphim urium /Am e s assay with the Chinese ham s ter liver S9 activation system (40 CFR 798.5265). 4. Invivo m ouse m i cronucleus assay i.p. (40 CFR 798.5395). Tests 3. and 4. would trigger a lif etim e rodent bioassay if positive. For Environm ental toxicity and fate - Prior to Tier 1, the following physical-chem i cal properties need to be established: Melting point (40 CFR 796.1300) or boiling point (40 CFR 796.1220), water solubility (40 CFR 796.1840), log K ow(40 CFR 796.1550, 796.1570 or 796.1720), and vapor pressure (40 CFR 796.1950). Tier 1. The fish, daphnid, and algal (40 CFR 797.1050) acute toxicity tests from the aquatic base set of environm ental toxicity tests will be recomm e nded for aquatic exposures. The acute toxicity tests for fish (CFR §797.1400) and daphnids (CFR §797.1300) will be done using the flow-through m e thod with m easured concentrations, and effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations. If there is no significant risk from the PMN after the results of the fish and daphnid acute toxicity tests have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then go to Tier 2.

133Tier 2. Direct and Indirect Photolysis Screening Test (40 CFR 796.3765). If t1⁄2 < 2 days, go to Tier 3; if t1⁄2 > 2 days, then integrate the results into the risk assessm ent. If there is no significant risk from the PMN, then no further testing is recomm e nded. However, if there rem a ins a significant risk, then go to Tier 4. Tier 3a. If t1⁄2 < 2 days and photolysis products are known and/or identified, then assess photolysis products for environm ental risk. If there is no significant risk from the photolysis products from the PMN, then no further testing is recomm e nded. However, if there rem a ins a significant risk, then go to Tier 4. Tier 3b. If t1⁄2 < 2 days and photolysis products are not known and/or identifiable, then prepare a stock solution of PMN using the standard hum ic-containing solution described in the direct and indirect photolysis screening te st [40.796.3765 (b)(2) and (c)(2)], expose to sunlight for at least 6 half-lives (t1⁄2), and test photolysis products for toxicity with m o st sensitive species f r om environm ental base set. For exam ple, if the m o st sensitive species f r om the environm ental base set has an EC50 value = 2.0 m g PMN/L (based on 100% active ingredients [AI]), then prepare a 5.0 m g PMN per liter stock solution based on 100% AI using the standard hum iccontaining solution. This stock solution is exposed to sunlight for at least 6 half-lives to ensure that all of the PMN has been photolyzed, and then this stock solution is used to retest the m o st sensitive aquatic species to determ ine if the photolysis products of the PMN are m o re or less toxic that the PMN. Integrate these toxicity results into the risk assessm ent. If there is no significant risk from the PMN, then no further testing is recomm e nded. However, if there rem a ins a significant risk, then go to Tier 4. Tier 4. Test for aerobic biodegradability according to any one of the following test guidelines (listed in order of preference): Modified Sturm Test 40 CFR 796.3260 Closed Bottle Test 40 CFR 796.3200 Modified OECD Screening Test 40 CFR 796.3240 Modified MITI Test (I) 40 CFR 796.3220 Modified AFNOR Test 40 CFR 796.3180 Integrate the results f o r aerobic biodegradability into the risk assessm ent. If there is no significant risk from the PMN, then no further testing is recomm e nded. However, if there rem a ins a significant risk, then do chronic toxicity testing for fish and daphnids: Fish chronic toxicity testing, i.e., fish early life stage (ELS) toxicity testing (CFR §797.1600), with flow-through m e thods; m easured concentrations; effective concentrations

134based on 100% active ingredients (AI) and m ean m easured concentrations; and statistical analysis of effective concentrations at days 7, 14, 21, and 28. Daphnid chronic toxicity testing (C FR §797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% active ingredients (AI) and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21. For Terrestrial ecosystem sThe terrestrial base set of environm ental toxicity tests (i.e., the early seeding growth test, the earthworm toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded f o r terrestrial exposures. Chronic toxicity testing fo r terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay. References: Am erican Dye Manufacturers Institute, Inc. (ADMI), 1974, "Dyes and the Environm ent: Reports on Selected Dyes and Their Effects," Vol. II, ADMI, New York. Au W , Pathak S, Colie CJ, Hsu TC. 1978. Cytogene tic toxicity of Gentian Violet and Crystal Violet on m amm a lian cells invitro

. Mutation Research58:269-276. Au W , Butler MA, Bloom SE, Matney TS. 1979. Further study of the genetic toxicity of Gentian Violet. Mutation Research66:103-112. Auer, C. M., Nabholz, J. V. and Baetcke, K. P., 1990, "Mode of Action and the Assessm ent of Chem ical Hazards in the Presence of Lim ited Data: Use of Structure-Activity Relationships (SAR) under TSCA, Section 5," Environm ental Health Perspectives , Vol. 87, pp. 183-197. Kier LD, Brusick DJ, Auletta AE. 1986. The Salm onella typhim urium /m amm a lian m i crosom al assay. A report of the U.S. Environm ental Protection Agency Gene-Tox Program

. Mutation Research168:69-240. Kirk-Othm e r, Encyclopedia of Chem ical Technology , 3rd edition, vol. 23, pages 399-412. Little, L. W

. and Lam b J. C., III, 1972. Acute Toxicity of 46 Selected Dyes to the Fathead Minnow, Pim e phales prom elas ,Final Report to the Am erican Dye Manuf acturers Institute,Inc., UNC W a stewater Research Center, Departm e nt of Environm ental Sciences and Engineering, School of Public Health, University of North Carolina, Chapel Hill, NC. Littlefield NA, Blackwell B-N, Hewitt CC, Gaylor DW . 1985. Chronic toxicity and carcinogenicity studies of Gentian Violet inm i ce. Fundam e ntal and Applied Toxicology5:902- 912.

135Mortelm ans K, Haworth S, Lawlor T. 1986. Salm onellam utagenicity tests: II. Results from the testing of 270 chem icals. Environm ental Mutagenesis 8(7):1-119. Nabholz, J. V., 1990. The OTS PMN ECOTOX Data Base: a Confidential Business Inform ation (CBI) Collection of Environm ental Toxicity Data from New Chem ical Subm issions Under Sec. 5

. Unpublished, Environm ental Effects Branch, Health and Environm ental Review Division (TS-796), U. S. Environm ental Protection Agency, W a shington, DC. NCTR Technical Report for Experim e nt Num b ers 305,354,355. Three-generation reproduction and toxicity studies of Gentian Violet in Fi scher 344 Rats. National Center for Toxicological Research. Jefferson, Arkansas. Sept. 1988. NCTR Technical Report for Experim e nt Num b er 355. Developm ental toxicity study of Gentian Violet in Fischer 344 Rats. National Center fo r Toxicological Research. Jefferson, Arkansas. Sept. 1988. NTP. 1985. National Toxicology Program . C.I. Basic Red 9 m onochloride. NTP TR-285. Sigm an, C. C., Helm s, C. T., Papa, P. A., Atki nson, D. L., Doeltz, M. K. and W i nship-Ball, A., Jan. 1983. Anthraquinone Dyes and Related Chem icals: Review and Assessm ent of Potential Environm ental and Health Aspects . Final Report to the Dyes Environm ental and Toxicology Organization, Inc., SRI International, Menlo Park, CA. Sokolowska-Pituchowa J, Kowalczkowa J, Kus J, Piotrowski J, Sawicki B. 1965. Teratogenic effect of Malachite Green in experi m e ntal anim als. Folia Biologica 13:311-315. Tonogai, Y., Ito, Y., Iwaida, M., Tati, M., Ose, Y. and Sato, T., 1979. Studies on the Toxicity of Coal-Tar Dyes. II. Exam ination of the Biological Reaction of Coal-Tar Dyes to Vital Body. The Journal of Toxicological Sciences , Vol. 4, pp. 211-219. July, 1994.

136Category: Vinyl Esters Hum an Health Environm ental Toxicity (See “Esters”) Definition: A carboxylic acid ester with at least one vinyl group (CH 2 =CH-) attached to an organic acid radical (RCOO-). A sim p le vinyl ester is vinyl acetate:

CH 3 COOCH=CH 2

Note: Certain carboxylic acid m o ieties them selves m ay exert toxicity such that there are concerns for effects in addition to those identified for the vinyl ester class.
Haz a rd Concerns: Oncogenicitybased on data from two bioassays: a 2-yr drinking water study of vinyl acetate in rats [significantly increas ed incidences of adenocarcinom as of the uterus and com b ined C-cell adenom as and carcinom as in high-dose (2,500 m g /l) fem a le rats] and a chronic inhalation study of vinyl acetate in rats and m i ce (laryngeal/nasal tum o rs in rats, bronchiolar/alveolar adenom as in lungs of m i ce)

. There are supporting data from 27-m o and 81- wk inhalation studies of acetaldehyde in rats and ham s ters, respectively. Furtherm ore, m

utagenicity data on these analogues support a cancer concern. Neurotoxicitybased on data on an analogous PMN com pound in rats plus suggestive data on vinyl acetate. Reproductive toxicitybased on a two-generation study of vinyl acetate in rats. Environm ental toxicity(see "Esters" category). Boundaries: Other than a requirem e nt for a vinyl group and an acid group, none. General Testing Strategy (1) Up-front glove perm eation study if there is a risk of any toxicity through derm al exposure. (2) Hydrolysis test (both acid- and enzym e -catalyzed) to determ ine the rate at which acetaldehyde, a potentially toxic m e tabolite, is released. [The requirem e nt to m easure acetaldehyde release is based on the assum p tionthat acetaldehyde is the presum ed "ultim ate carcinogen" of vinyl esters. This requirem e nt will be revisited as m o re decisive data becom e available. The hydrolysis data should be helpful in determ ining the m o st appropriate route of adm i nistration in the f o llowing tests.] (3) A 90-day neurotoxicity test in rats that includes a functional observational battery, m o tor activity tests, and neuropathology exam inations. Also to be included, to address reproductive toxicity concerns, are the following adjuncts to the subchronic test:

137(a) W e ights of the testes, epididym ides (total and cauda), pituitary, sem i nal vesicles (with coagulating glands), prostate, ovary, and uterus are to be recorded at the tim e of sacrifice. (b) Histopathology of the testes is to be done on the m a les at the tim e of sacrifice. Particular attention is to be directed toward achieving satisf actory quality f r om f

i xation and em bedding, and preparations are to follow the recomm e ndations of Russell et al. (1990). Histologic analyses are to include evaluations of the sperm a togenic cycle; i.e., the presence and integrity of the 14 cell states. These evaluations would follow the guidance provided by Russell et al. (4) A standard developm ental toxicity study in rabbits and rats. (5) A 2-yr bioassay in rats and m i ce to address cancer concerns. (6) Environm ental toxicity testing: see "Esters" category.

References.

Russell LD, Ettlin RA, Sinha Hikim AP, Cl egg ED. 1990. Histological and histopathological evaluation of the testis. Cache River Press, Clearwater, FL. August, 1992

138Category: Vinyl Sulfones Hum an Health Environm ental Toxicity Definition. Any m o lecular structure with a vinyl sulfone group is considered to be a m e m b er of the class:
O 5 -S-CH=CH 2

5 O In addition the following is an exam ple of a functional group that generates a vinyl sulfone under certain conditions. 

### $ -Sulfatoethyl-sulfonyl groups are typical vinyl sulfone precursors. Structures bearing this group are also m e m b ers of the class:
O 5

- -S-CH 2 -CH 2 -OSO 3

5 O Typically, the new chem ical of concern is a fiber-reactive dye bearing one or m o re vinyl sulfones, or vinyl sulfone precursors. Occupati onal exposure to workers via inhalation/ingestion of the powdered dye m ay pose a potential health risk. Under the conditions in a dye bath a vinyl sulfone precursor-containing dye generates the vinyl sulfone. In this scenario the only potential risk is to those who drink contam inated drinki ng water. However, following covalent binding of vinyl sulfone to textile fibers, survival of unbound vinyl sulfone substituents in the dye bath is low (< 10%). The m a jority of unbound vinyl sulfone is hydrolyzed to 

### $ -hydroxyethylsulfone. Unless releases to water are extrem ely high, vinyl sulfones are not expected to pose a significant risk to hum an health. Presently, there are no hum an health concerns for 

### $ -hydroxyethylsulfone.
Haz a rd Concerns. For those who inhale or ingest a vinyl sulfone, there is an oncogenicity concern and m utagenicity concerns based on the potent m utagenicity of vinyl sulfone (VS) and m e thylvinyl sulfone (MVS). VS and MVS are m utagenic in the L5178Y TK+/

- m ouse lym phom a gene m utation assay invitro

. Evaluation of sm all colonies indicates that genotoxicity is due to a clastogenic m echanism

. This is confirm e d by evaluating the lym phom a cells for chrom o som e aberrations and m i cronuclei. MVS also induces effects upon the spindle apparatus in Chinese ham s ter lung cells invitro , indicating an aneugenic (aneuploi dy-inducing) activity. A directacting Michael addition-type reaction m ay be the m echanism of action. Although MVS and divinyl sulfone (DVS) are both reported as negative invivo in the m ouse m i cronucleus assay (inconclusive, only m a les tested), and in the dom inant lethal assay, the Agency has determ ined that these negatives are not sufficient to re m ove concern for vinyl sulfone-containing new chem icals as potential m utagens and carcinogens. There are ecotoxicity concerns for VS only, based on concerns for electrophiles.

139Boundaries. There are at present no boundary conditions, related to health effects, for structures containing vinyl sulfones. Nearly all new chem icals in the category have been water-soluble, fiber-reactive dyes with m o lecular weights <1,000. The boundary conditions for ecotoxicity effects are m o lecular weights < 1000 and log Kow< 8. General Testing Strategy The New Chem icals Program considers the following tests to be appropriate to address health and ecotoxicity concerns: 1. Mouse lym phom a assay with evaluation of sm all colonies (40 CFR 798.5300). The test would trigger a lif etim e rodent bioassay if positive. 2. Base-set ecotoxicity testing to incl ude fish (40 CFR 797.1400), daphnids (40 CFR 797.1300) and algae (40 CFR 797.1050). The acute toxicity tests for fish and daphnids will be done using the flow-through m e thod with m easured concentrations, and effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations. If there is insufficient knowledge about the water solubility of the dye, then it should also be m e asured (40 CFR 796.1840 or 796.1860). If there is no significant risk from the PMN after the results of the fish and daphnid acute toxicity and algal toxicity tests have been integrated into the risk assessm ent, then no further testing is recomm e nded. However, if there is a significant risk, then do environm ental fate testing as outlined in 3. 3. Environm ental f a te testing f o r releases to water. Tier 1. Direct and Indirect Photolysis Screening Test (40 CFR 796.3765). If t1⁄2 < 2 days, go to Tier 2; if t1⁄2 > 2 days, go to Tier 3. Tier 2a. If t1⁄2 < 2 days and photolysis products are known and/or identified, then assess photolysis products for environm ental hazards. Tier 2b. If t1⁄2 < 2 days and photolysis products are not known and/or identifiable, then prepare a stock solution of the PMN chem ical using the standard hum ic-containing solution described in the direct and indirect photol ysis screening test [40 CFR 796.3765 (b)(2) and (c)(2)], expose to sunlight for at least 6 half-liv es (t1⁄2), and test photolysis products for toxicity with the m o st sensitive species f r om the environm ental base set.

140Tier 3. Test for aerobic biodegradability according to any one of the following test guidelines: Modified Sturm Test 40 CFR 796.3260 Closed Bottle Test 40 CFR 796.3200 Modified OECD Screening Test 40 CFR 796.3240 Modified MITI Test (I) 40 CFR 796.3220 Modified AFNOR Test 40 CFR 796.3180 In addition, if appropriate, physical/chem i cal properties including: Log K ow(40 CFR 796.1550, 796.1570, or 796.1720) Vapor pressure (40 CFR 796.1950) Boiling point (40 CFR 796.1220) Melting point range (40 CFR 796.1300) If the PMN chem ical passes the aerobic biodegradability test according to the criteria set forth in the guidelines, no further testing m ay be recomm e nded. A new risk assessm ent is conducted to determ ine if chronic ecotoxicity testing is warranted. 4. Fish chronic toxicity testing, i.e., fish ear ly life stage (ELS) toxicity testing (40 CFR 797.1600), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% AI and m ean m easured concentrations; and statistical analysis of effective concentrations at days 7, 14, 21, and 28. Daphnid chronic toxicity testing ( 40 CFR 797.1330), with flow-through m e thods; m easured concentrations; effective concentrations based on 100% AI and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21.
5. Ecotoxicity testing for release to terrestrial ecosystem s. The terrestrial base set of environm ental toxicity tests (i.e., the early seedling growth test, the earthworm toxicity test and the soil m i crobial comm unity bioassay) will be recomm e nded f o r terrestrial exposures. Chronic toxicity testing fo r terrestrial organism s include: the plant whole life cycle test, the plant uptake test, and the soil m i crobial comm unity bioassay.

References: Dearfield, KL, K Harrington-Brock, CL Doe rr, JR Rabinowitz, and MM Moore (1991). Genotoxicity in m ouse lym phom a cells of chem icals capable of Michael addition. Mutagenesis 6: 519-525.

141Heyna, J (1963). Reactive dyes containing vinylsulfonyl groups. Angew . Chem. Internat. Edit. 2(1): 20-23. Shelby, MD, GA Gutierrez-Espeleta, W M Generoso, and AF McFee (1991). Mouse dom inant lethal and bone m arrow m i cronucleus studies on m e thyl vinyl sulfone and divinyl sulfone. Mutation Research 250: 431-437.
W arr, TJ, JM Parry, RD Callander, and J Ashby ( 1990). Methyl vinyl sulfone: A new class of Michael-type genotoxin. Mutation Research 245: 191-199. Septem ber, 1988; revised June, 1994

142Category: Soluble complexes of Z i nc Environm ental Toxicity Definition. This category includes all organic and inorganic soluble com p lexes of zinc, for exam ple, zincates, organozincs, such as, dim e thylzinc. Not included in this category are zinccom p lexed acid dyes and strong ion pairs between zinc and anionic surfactants. There are no toxicity data for soluble com p lexes of zinc, therefore, the toxicity data for the free Zn ++ ion will be used to predict aquatic toxicity.
Haz a rd Concerns. Zinc produces acute toxicity to freshwater aquatic organism s over a range of concentrations from 90 to 58,100 ug/L and produces chronic toxicity over a range from 47 to 852 ug/L. The acute to chronic ratio (ACR) f o r Zn has been set at 3 in the water quality criteria (W QC) docum ent. Zinc concentrations from 30 to 21,600 ug/L have been shown to reduce the growth of various aquatic plants and algae appear to be the m o st sensitive group. Bioconcentration factors (as logarithm s ) for fres hwater fish ranged from 1.7 to 2.6, and from 2.0 to 3.1 for freshwater invertebrates. Zinc is an essential nutrient trace elem ent which can be toxic at higher concentrations. Wh ile acute toxicity is affected by hardness, chronic toxicity is not affected. The freshwater criteria at an average hardness of 100 m g /L are 0.120 ug/L for acute exposures and 0.110 ug/L for chronic exposures; the saltwater criteria are 0.095 ug/L for acute exposures and 0.086 ug/L for chronic exposures. These criteria are based on toxicity inform ation for the following Zn salts: sulfate, chloride, phosphate, and nitrate. General Testing Strategy To address ecotoxicity concerns, base set acute aquatic toxicity testing (algae: static m e thod, daphnid and fish: flow-through m e thod, all m easured concentrations). April, 1990

143Category: Z i rconium Compounds Environm ental Toxicity Definition. This category includes inorganic salts of zirconium (Zr), com p lexes between Zr and organic acids, and organoZr com pounds, i.e., Zr covalently-bonded with carbon. For exam ple, som e inorganic Zr salts include: Zr sulfate, Zr oxychloride, and Zr tetrachloride. Not included in this category are dyes com p lexed with Zr.
Haz a rd Concerns. Soluble salts of Zr are known to be m oderately toxic to green algae and fish based on exposures in soft water and in term s of m g Zr/L. Toxicity inform ation are only available for Zr sulfate, Zr tetrachloride, and Zr oxychloride. The toxicity profile for Zr based on available toxicity data, m g Zr/L (ppm Zr), and m oderate hardness (i.e., 150.0 m g /L as CaCO 3 ) is: fish 96-h LC50 = 58.0 (n=3) green algal 96-h EC50 = 2.6 Zirconium is m o re toxic in soft water than hard water. Acute toxicity to fish have been shown to increase 13 tim es as hardness decreases from 400.0 to 20 m g /L. Boundaries. The toxicity of Zr com pounds depends on the their physical/chem i cal properties and the hardness of receiving waters. The toxicity of Zr salts and Zr com p lexes with organic acids are expected to be related to their water solubility and MW

. The toxicity of organoZr com pounds are also expected to be related to their octanol/water partition coefficient (Kow). The m o st im portant property determ ining the toxicity of Zr com pounds is water solubility. W a ter solubility cannot be estim ated accurately and has to be m easured. The water solubility of organoZr com pounds is expected to decrease as Kow increases. There is no lower bound for Kow and the upper bound cannot be determ ined at this tim e since the Kow fragm e nt-constant for Zr is not known. In addition to solubility, MW is also an im portant boundary. Com pounds with MW s > 1000 are not expected to be absorbed by aquatic organism s even if they are water soluble. Therefore, only Zr com pounds with MW s < 1000 are expected to be toxic. General Testing Strategy
I. Release to Aquatic Ecosystem s Tier 1. The aquatic base set of environm ental toxicity tests will be recomm e nded f o r aquatic exposures. The acute toxicity tests for fish (40 CFR §797.1400) and daphnids (40 CFR §797.1300) will be done using the flow-through m e thod; effective concentrations will be based on 100% active ingredients (AI) and m ean m easured concentrations; the total organic carbon (TOC) concentration of dilution water in the control m u st be less than 2.0 m g TOC/L; TOC m

u st be m easured in the control just prior to the start of the test; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested com pound; solvent can be used to assist the com pound to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water

144solubility of the com pound beyond its aqueous solubility lim it; and hardness of dilution water

3 . If toxicity m itigation testing is done with hum ic acid, then the static m e thod with nom inal concentrations will be recomm e nded. has to be less than 180.0 m g /L as CaCO
The algal toxicity testing (40 CFR §797.1050), should be done with the static m e thod; effective concentrations based on 100% AI and m ean m easured concentrations; statistical analysis of effective concentrations at 24, 48, 72, and 96 hours; test m e dium with no m o re than 0.300 m g /L EDTA as a final concentration; the TOC of the test/growth m e dium should be less than 2.0 m g TOC/L; TOC should be m easured just prior to the start of the test; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested com pound; and solvent can be used to assist the com pound to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the com pound beyond its aqueous solubility lim it. If toxicity m itigation testing is done with hum ic acid, then nom inal concentrations will be recomm e nded. If there is no significant risk from the Zr com pound after the results of the environm ental base set have been integrated into the risk assessm ent, then no f urther testing will be recomm e nded. However, if there is a significant risk, then go to Tier 2. Tier 2. Fish chronic toxicity testing, i.e., fi sh early life stage (ELS) toxicity testing (40 CFR §797.1600), with the flow-through m e thod; eff ective concentrations based on 100% AI and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, 21, and 28; the TOC of dilution water in the control should be less than 2.0 m g TOC/L; TOC should be m easured in the controls just prior to and during the test; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested com pound; solvent can be used to assist the com pound to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the com pound beyond its aqueous solubility lim it; and hardness of dilution water has to be less than 180.0 m g /L as CaCO 3 . Daphnid chronic toxicity testing (40 CFR §797.1330), with the flow-through m e thod; effective concentrations based on 100% AI and m ean m easured concentrations; statistical analysis of effective concentrations at days 7, 14, and 21; the TOC of dilution water in the control should not exceed 2.0 m g TOC/L; TOC m u st be m e asured in the controls just prior to and during the test; the highest treatm e nt concentration on a nom inal-basis should not exceed the aqueous solubility lim it of the tested com pound; solvent can be used to assist the com pound to reach its aqueous solubility lim it quicker, but cannot be used to artificially enhance the water solubility of the com pound beyond its aqueous solubility lim it; and hardness of dilution water has to be less than 180.0 m g /L as CaCO 3 . II. Release to Terrestrial Ecosystem s : The terrestrial base set of environm ental toxicity tests will be recomm e nded for terrestrial exposures. The terrestrial base set includes: the early seeding growth test (40 CFR 797.2800), the earthworm toxicity test (40 CFR.795.150), the soil m i crobial comm unity bioassay (40 CFR 797.3700), and the avian acute oral toxicity test (40 CFR 797.2175). Chronic toxicity testing for terrestrial organism s include: the plant whole life cycle test (40 CFR 797.xxxx), the plant uptake test (40 CFR 797.2850), and the avian reproduction test (40 CFR 797.2130 for bobwhite quail or 40 CFR 797.2150 for m a llard duck). June, 1992
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