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# REVISED

NTAN HPV TEST PLAN

Submitted to the U.S. Environmental Protection Agency

By

Akzo Nobel Functional Chemicals LLC August 2011 Akzo Nobel Services Inc. HSE Department 525 W Van Buren Street Chicago, Illinois 60607-3835 United States T +1 312 544 7000 F +1 312 544 7125 www.akzonobel.com/tePage 2 of 10 

### TABLE OF CONTENTS

Summary ...............................................................................................Page 3

1.0 Introduction ................................................................................... Page 4

2.0 Evaluation of Existing Data and Proposed Testing

Physical/Chemical Properties............................................................Page 4 Environmental Fate..........................................................................Page 6 Aquatic Toxicity.............................................................................. Page 7 Acute Toxicity.................................................................................Page 7 Repeated Dose.............................................................................. Page 9 Reproductive/Developmental Toxicity................................................... Page 9 Mutagenicity................................................................................... Page 9

3.0 Use and Exposure Information ........................................................ Page 10

Appendix A : NTAN HPV Robust Summaries (IUCLID5)Page 3 of 10 SUMMARY

Akzo Nobel Functional Chemicals LLC has sponsored acetonitrile, 2,2’2’’-nitrilotris- (CAS# 7327- 60-8), also known as NTAN, in the U.S. EPA High Production Volume (HPV) program. Experimental and modeling data exist on several SIDS endpoints for NTAN.

Robust summaries of available studies on NTAN are included in this submission (see Appendix A). Data on NTAN from the EPISuite computer model were used in the absence of experimental data for physical/chemical characteristics. The ECOSAR computer model was used in the absence of experimental data on ecotoxicity endpoints. The table below summarizes the endpoints of interest in the HPV program, the available data, and indicates if testing is proposed testing.

Endpoint Sufficient Data Available Testing Proposed Ph y sical/Chemical Characteristics Yes No Photodegradation Yes No H y drol y sis Yes No Biodegradation Yes No Transport Yes No Acute Fish Toxicity Yes No A cute Daphnia Toxicit y Yes No Acute Alga Inhibition Yes No A cute Toxicit y Yes No Genetic Toxicity Yes No Repeated Dose Yes No Reproductive Toxicity Yes No Developmental Toxicit y Yes NoPage 4 of 10 1.0 INTRODUCTION

Akzo Nobel Functional Chemicals LLC has sponsored NTAN (CAS# 7327-60-8) in the U.S. HPV program to assess its potential health and environmental hazards, including selected physical/chemical characteristics. In the absence of data on NTAN, U.S. EPA validated computer models were used to generate estimated values.

In the previous version of the test plan (February 2009) it was proposed to perform vapor pressure (OECD 104), water solub ility (OECD 105), combined repeat dose/reproductive/developmental and genetic toxicity (OECD 422) studies on NTAN. These studies are now finalized.

Additional studies were perfor med to comply with the REACH (Registration Evaluation Authorization and Restriction of Chemicals) legislation as follows.

Physical/chemical characteristics; density (OECD 109), particle size distribution (NENISO 13320-1) and auto fla mmability (EU method A.16).

Environment: Biodegradation in water (OECD 301D), short-term toxicity to aquatic invertebrates (OECD 202) and toxicity to aquatic algae and cyanobacteria (OECD 201).

All data mentioned above, in addition to existing data on phys-chem, acute toxicity and irritation obtained during data gathering under REACH ha s been summarized in IUCLID5 and included in the test plan.

2.0 EVALUATION OF EXISTING DATA AND PROPOSED TESTING

The available data on NTAN has been evaluated and summarized. Data on NTAN from the EPISuite computer model were used in the absence of experimental values for physical/chemical properties. If experimental data was not available the ECOSAR computer model was used to estimate aquatic toxicity. Data for acute mammalian toxicity was available from Akzo Noble Functional Chemicals LLC sponsored studies as well as from submissions made to the U.S. EPA under TSCA section 8(e). Data is now available from studies that were proposed in the previous version of the test plan and studies performed to comply with the REACH legislation. Robust summaries of the studies are included in this subm ission. The Klimisch reliability code was used to determine the reliability of the available stud ies. A literature search of online databases included: TOXNET (TOXLINE, HSDB, IRIS, GENETOX, CCRIS), PubMed, RTECS, European Chemicals Bureau (IUCLID), SIDS, EPA HPV, NTP, NIEHS, IARC, CIS (Aquire, Datalog, Envirofate, BIODEG), EPA TSCA 8(e), TSCATS and STN. Studies identified included acute toxicity studies submitted to U.S. EPA under TSCA section 8(e).

Physical/Chemical Properties:

NTAN is acetonitrile, 2,2’2”-nitrilotris- with th e CAS# of 7327-60-8. The molecular formula of CAS# 7327-60-8, verified by the National Chemical Inventories database, is C 6 H 6 N 4 , (National Chemical Inventories, 2008). Based on the molecular formula the molecular weight of NTAN is 134. The structure of NTAN is provided below.Page 5 of 10 C N

N

C
N C

N

MolWt: 134.14 C6 H6 N4007327-60-8 Acetonitrile, 2,2',2''-nitrilotrisAll data for physical/chemical properties are provided in the table below. The estimated values were calculated using the CAS number as input in the US EPA EPISuite model.

NTAN PHYSICAL/CHEMICAL DATA *
Parameter Guideline(s) Result Sponsor/Lab/Year Validity Melting temperature EC A.1 OECD 102 125.8– 127.7°C. Akzo/Akzo/2006 (2) valid with
restrictions Melting temperature EC A.1 OECD 102 128°C. Akzo/Akzo/2003 (2) valid with
restrictions Melting temperature - 92.78°C EPISuite (2) valid with
restrictions Boiling temperature EC A.2 OECD 103 > 120°C, the substance evaporates during melting and before boiling Akzo/Akzo/2003 (2) valid with
restrictions

Boiling temperature - 321.24°C EPISuite (2) valid with
restrictions Relative density EC A.3 OECD 109 1.298 Akzo/Akzo/2010 (1) valid without
restrictions
Particle size
distribution NEN-ISO 13320-1 Percentile Mean

D10 > 92 — < 120 μm

D50 > 185

— < 246 μm

D90 > 304

— < 371 μm Akzo/Akzo/2010 (1) valid without
restrictions

Vapour pressure EC A.4 OECD 104 0.000104 Pa at 25 °C,
7.8x10 -6 mmHg Akzo/ABC/2009 (1) valid without
restrictions Vapour pressure EC A.4 OECD 104 0.001 mBar at 80°C The substance starts to decompose above 80°C, 7.5x10 -4 mmHg Akzo/LTP/2006 (1) valid without
restrictions

Vapour pressure - 3.17 x 10 -7 mmHg Daubert & Danner
1989 (4) not assignable

Vapour pressure - 6.17x10 -5 mmHg EPISuite (2) valid with
restrictions
Partition coefficient - -1.39 EPISuite (2) valid with
restrictions Water solubility EC A.6 OECD 105 > 3.02 < 3.1 g/l at 20°C, pH 6.9 Akzo/ABC/2009 (1) valid without
restrictions Water solubility - 1x10 6 mg/l EPISuite (2) valid with
restrictions Surface tension EC A.5 OECD 115 72.6 nM/m at 22°C, Akzo/Akzo/2006 (1) valid withoutPage 6 of 10 conc. 1.06 g/L restrictions Auto flammability EC A.16 The substance did not show self-heating up to its melting point
which means that it
does not have a selfignition temperature. Akzo/Akzo/2010 (1) valid without
restrictions

Flammability EC A.10 Not flammable Akzo/Akzo/2002 (2) valid with
restrictions Pyrophoric properties EC A.13 Not pyrophoric Akzo/Akzo/2002 (2) valid with
restrictions

Explosive properties EC A.14 Not explosive Akzo/Akzo/2002 (2) valid with
restrictions Oxidizing properties EC A.21 Not oxidizing Akzo/Akzo/2002 (2) valid with
restrictions * Key value is printed bold

The substance has no self-heating or self-reactive properties and it is not produced nor transported as an aerosol or as a gas under pressure. NTAN has no physical/chemical properties that require classification and labeling.

Recommendation: No additional testing is proposed.

Environmental Fate:

Modeling results for NTAN estimate the atmosphe ric half-life to be 25.924 hours indicating a moderate rate of degradation.

The EPISuite model indicates that a hydrolysis ha lf-life cannot be estimated for NTAN at 25°C.

The EPISuite Level III fugacity model was used to estimate the distribution of NTAN. The modeling results indicate that NTAN is primarily distributed to water and soil.

The estimated log Koc is 1.15 L/kg or 0.87 L/kg based on MCI or Log Kow, respectively. The values were calculated with KOWWINv2.00. The estimated log Koc values are less than the threshold value of 3, indicating no adsorbing potential to soil for this compound.

Modeling data predicts that NTAN is not biodegradable. The Re ady Biodegradability Prediction from BIOWIN v4.10 was “NO”. According to the BIOWIN Users’ Guide, “The criteria for the YES or NO prediction are as follows: if BIOWIN3 (ultimate survey model) result is “weeks” or faster (e.g. days or days to weeks) AND BIOWIN5 (MITI linear model) >= 0.5, then the prediction is YES (readily biodegradable)...If this condition is not satisfied, the prediction is NO (not readily biodegradable).” The BIOWIN3 (ultimate survey model) for NTAN predicted months and BIOWIN5 (MITI linear model) predicted 0.59. Since both criteria were not met, the prediction was “NO” (not readily biodegradable).

These predictions are confirmed by a closed bottle test (Van Ginkel 2010). NTAN is not biodegraded in the prolonged Closed Bottle test. NTAN should therefore not be classified as readily biodegradable. The lack of biodegradation in the Closed Bottle test does not mean that NTAN is persistent in nature because the stringency of the test procedures could account for the recalcitrance in the Closed Bottle test.Page 7 of 10 Environmental release of NTAN is expected to be minimal as closed systems are employed during manufacture. Furthermore, NTAN is used as a chemical intermediate and is completely consumed in the manufacturing process of final products.

Recommendation: No additional testing is proposed.

Aquatic Toxicity:

An acute toxicity study to aquatic invertebrates and a toxicity study to aquatic algae are available. The ECOSAR computer model was also used to assess the aquatic toxicity of NTAN. The values are provided in the table below.

Ecotoxicity Data on NTAN *
Parameter Guideline(s) Result Sponsor/Lab/Year Validity Fish LC50 - 5460 mg/L ECOSAR (2) valid with restrictions

Invert. LC50 OECD 202 48h EC50 > 1000 mg/L
48h NOEC ≥ 1000 mg/L Akzo/Akzo/2010 (1) valid without
restrictions Invert LC50

- 260 mg/l ECOSAR (2) valid with restrictions

Algae EC50 OECD 201 72 h NOEC 171 mg/L 72 h LOEC 309 mg/L 72 h EC10 652 mg/L 72 h EC50 > 1000 mg/L Akzo/Akzo/2010 (1) valid without
restrictions

Algae EC50 - 152 mg/L ECOSAR (2) valid with restrictions Algae ChV - 15 mg/L ECOSAR (2) valid with restrictions Log BCF - 0.5 ECOSAR (2) valid with restrictions * Key value is printed bold

Recommendation: No additional testing proposed.

Acute Toxicity:

Several acute oral toxicity studies are available on NTAN. Valid studies indicate the acute oral toxicity of NTAN to be approximately 200 mg/kg. The table below contains available acute oral studies on NTAN with the Klimisch reliability code.

SPONSOR/LAB/YEAR RESULTS RELIABILITY Monsanto/Younger/1965 LD50 (rats) = 150 mg/kg; clinical signs (4) not assignable Monsanto/Younger1966 LD50 (rats) = 198 mg/kg; clinical signs (4) not assignable Kodak/1971 LD50 (rats) = 50 mg/kg; clinical signs (4) not assignable Kodak/1971 LD50 (mice) = 50-100 mg/kg (4) not assignable Dow/1967 LD50 (rats) = 50-250 mg/kg (2) valid with restrictions

Akzo/ToxMonitor/2001 LD50 (rats) <200 mg/kg (Lot #3554B); clinical signs (1) valid without restrictions

Akzo/Covance/2001 LD50 (rats) >50<200 mg/kg (Batch #3554B); clinical signs (1) valid without restrictions

Akzo/ToxMonitor/2001 LD50 (rats) >200 mg/kg (recrystallized); clinical signs (1) valid without restrictions

Akzo/Covance/2001 LD50 (rats) ~=200 mg/kg (recrystallized); clinical signs (1) valid without restrictions

Akzo/ToxMonitor/2001 LD50 (rats) <200 mg/kg (lab washed); clinical signs (1) valid without restrictionsPage 8 of 10 Akzo/Covance/2001 LD50 (rats) >200 mg/kg (lab washed); clinical signs (1) valid without restrictions

Akzo/ToxMonitor/2001 LD50 (rats) >200 mg/kg (Lot #01013N34603; clinical signs (1) valid without restrictions

Akzo/Covance/2001 LD50 (rats) >50<200 mg/kg (Lot #01013N346403); clinical signs (1) valid without restrictions

Four acute dermal studies were identified from database searches. Three of the studies were submitted to U.S. EPA under TSCA section 8(e). The results from one study indicate the LD50 to rabbits is between 794 and 1260 mg/kg. Animals exhibited clinical signs of toxicity. In the second study, the LD50 was reported to be greater than 2000 mg/kg with no signs of clinical toxicity. The third study showed that the acute dermal LD50 in guinea pigs was greater than 1000 mg/kg. In a fourth study rabbits were exposed to 200 mg/kg bw. None of the animals died. Detailed study design and results were not provided in all reports. However, there are sufficient details to estimate the acute dermal toxicity of the HPV substance. Furthermore, occupational exposure is expected to be minimal as closed systems are employed during manufacture and processing.

SPONSOR/LAB/YEAR RESULTS RELIABILITY

Monsanto/Younger/1965 LD50 (rabbits) >794<1260 mg/kg; clinical signs (4) not assignable

Monsanto/Younger/1966 LD50 (rabbits) >2000 mg/kg; No clinical signs (4) not assignable

Kodak/1971 LD50 (guinea pig) >1000 mg/kg; No clinical signs (4) not assignable

Dow/1978 LD0 (rabbits) = 200 mg/kg (2) valid with restrictions

Irritation studies indicate NTAN is not irritating to ra bbit skin and slightly irritating to rabbit eyes.
SPONSOR/LAB/YEAR RESULTS RELIABILITY

Monsanto/Younger/1965 Not irritating to skin (4) not assignable

Monsanto/Younger/1966 Not irritating to eyes (4) not assignable

Dow/1967 Not irritating to skin (2) valid with restrictions

Dow/1967 Not irritating to eyes (2) valid with restrictions

Dow/1978 Not irritating to skin (2) valid with restrictions

Dow/1978 Not irritating to eyes (2) valid with restrictions

The OECD toolbox was used to establish skin sensitising properties (QSAR). Based on the chemical structure of NTAN it is very unlik ely that NTAN has skin sensitising properties.

Recommendation: No additional testing proposed.Page 9 of 10 Repeated Dose:

A combined 28-day oral repeated dose study and reproductive/development study in rats was performed. The dose levels for this study were based on a 14-day range-finding study. Oral administration of NTAN to Sprague-Dawley rats at doses of 1, 10 and 45 mg/kg/day resulted in overt signs of parental toxicity at the 45 mg/kg/day dose including death, clinical signs of toxicity (including convulsions, labored breathing, etc.), decreased food consumption, body weight, body weight gain, and motor activity changes. It was noted that the clinical signs of toxicity occurred in females only (6/15) and the change in motor activity was only exhibited in males. It is difficult to discern whether the clinical signs of toxicity and change in motor activity are indicative of a neurotoxic effect as they occurred at a dose level which produced overt systemic toxicity (death). Interpretation of these types of indicators “as neurotoxic effects is dependent on the dose at which such changes occur and the possibility that damage to other organ systems may contribute to or cause such changes indirectly”.

Treatment-related gross necropsy findings were low/singular in incidence, limited to the 45 mg/kg group and consisted of red fluid/blood in the cranial cavity/cranium (observed in four animals that died spontaneously during the study), pigmentation of the brain, thymus and liver, urinary bladder dilatation and an enlarged thymus. No treatment-related histopathological changes were observed in the tissues examined from rats in the 45 mg/kg group; all findings were considered spontaneous or incidental changes commonly observed in rats of this age and strain. Based on the results of this study, the no-observed-adverse-effect level (NOAEL) for systemic toxicity of NTAN was 10 mg/kg/day.

Reproductive/Developmental Toxicity:

Overall mating, pregnancy, offspring survival and pup body weights were not notably affected by treatment with NTAN.

Gross external observations, following parturition on lactation day 0, were low in incidence and were noted in 4-6 fetuses in each treatment group, including the vehicle control group. All gross external morphology observations were common observations likely associated with delivery and consisted mainly of bruising, pale skin and a lesion/scab on the neck. None of the gross external observations were attributed to NTAN treatment. Visceral examinations were performed on dead pups, when feasible. No visible abnormalities were noted during the visceral examinations. The information described above indicates that NTAN did not induce reproductive or developmental toxicity at a dose level up to 45 mg/kg bw when given during 2 weeks prior to mating, during mating and during the complete pregnancy period.

The NOAEL for reproductive/developmental toxicity was at least 45 mg/kg/day.

Recommendation: No additional testing proposed.

Mutagenicity:

Two in vitro genotoxicity stud ies were performed, an Ames test and a Chromosome Aberration test in CHO cells. Both in vitro genotoxicity studies, viz. the Ames test and the chromosome aberration test showed that NTAN was not mutagenic.

Recommendation: No additional testing proposed.Page 10 of 10 3.0 USE AND EXPOSURE INFORMATION

NTAN is used in an industrial setting as an isol ated chemical intermedia te which is completely consumed in the reaction process. There is limited opportunity for occupational exposure or environmental release because closed systems are employed and the chemical is stored in super sacks which are loaded from and discharged directly into reactor vessels without direct contact by workers. NTAN is not used in consumer products. Environmental exposure and fate as well as occupational exposure are summarized in Appendix B of this submission (submitted previously).