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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/te 
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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) 
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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 

Physical/Chemical Characteristics Yes No

Photodegradation Yes No 

Hydrolysis Yes No

Biodegradation Yes No 

Transport Yes No

Acute Fish Toxicity Yes No 

Acute Daphnia Toxicity Yes No

Acute Alga Inhibition Yes No 

Acute Toxicity Yes No

Genetic Toxicity Yes No 

Repeated Dose Yes No

Reproductive Toxicity Yes No 

Developmental Toxicity Yes No
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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 solubility (OECD 105), combined repeat 

dose/reproductive/developmental and genetic toxicity (OECD 422) studies on NTAN. These 

studies are now finalized. 

Additional studies were performed 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 flammability (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 has 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 submission. The Klimisch reliability code was used 

to determine the reliability of the available studies. 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 the CAS# of 7327-60-8. The molecular formula of 

CAS# 7327-60-8, verified by the National Chemical Inventories database, is C6H6N4, (National 

Chemical Inventories, 2008). Based on the molecular formula the molecular weight of NTAN is 

134. The structure of NTAN is provided below. 
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C N

N

C

N

C

N

MolWt: 134.14 C6 H6 N4 

007327-60-8 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 - 1x106 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 without 
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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 atmospheric half-life to be 25.924 hours indicating a 

moderate rate of degradation. 

The EPISuite model indicates that a hydrolysis half-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 Ready 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. 
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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 restrictions 
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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 rabbit 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 unlikely that NTAN has skin sensitising properties.

Recommendation: No additional testing proposed. 
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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 studies 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. 
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3.0 USE AND EXPOSURE INFORMATION

NTAN is used in an industrial setting as an isolated chemical intermediate 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).