Appendix A - acetonitrile, 2,2’2’’

‐ nitrilotris

‐ (CAS# 7327

‐ 60

‐ 8)

Robust summaries

‐ IUCLID5



### 4 Physical and chemical properties Endpoint summary: Physical and chemical properties UUID IUC5-c85658bc-64a8-440c-93d9-68ee4a4a0e2a Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-12 11:11:49 CET Remarks

Administrative Data

Discussion This substance is not produced nor transported as an aerosol nor as a gas under pressure. NTAN has no physical/chemical properties that need classification and labelling. 4.1 Appearance/physical state/colour Endpoint summary: Appearance/physical state/colour UUID IUC5-c9dce4cf-526a-4e2f-81d6-4a5e7bbef844 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:09:18 CEST Remarks

Administrative Data Short description of key information The substance is an off-white, odourless, organic powder (wet cake). Key value for chemical safety assessment Physical state at 20oC and 1013 hPa solid

Endpoint study record: 7327-60-8, Appearance/physical state/colour, Huybens, 2010, CoA, RS UUID IUC5-04964d1f-cfff-48fb-8b5e-e212375bf042 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:09:41 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Stud y period May 2006 Reliability 1 (reliable without restriction)

Rationale for reliabilit y incl. deficiencies Signed certificate of analysis. Data sourceReference Referenc

e type Author Year Title Bibliographi

c source Testing laboratory Report

no. Owner company Compan y study

no. Repor t date

study report Huyben s ICM 201 0 Certificat e of analysis NTAN Chelates (FC-C), Deventer, The Netherland

s Batch numbe r CFC 7538 AkzoNobe l Functional Chemicals bv 2010- 06-10

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test guideline Qualifier Guideline Deviations equivalent or similar to EPA OPPTS OPPTS 830.6303 (Physical State) equivalent or similar to EPA OPPTS 830.6304 (Odor) equivalent or similar to EPA OPPTS 830.6302 (Color) yes (visual) GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8

EC number 230-804-9 Details on test material Batch number CFC7538Purity 94,1 %W ater 3.2 %Expiration date April 2010 Results and discussion Physical state at 20oC and 1013 hPa solid Form other: Powder (wet cake) Colour off-white Odour odourless Substance type organic Overall remarks, attachments Overall remarks see:7327-60-8, Densit y , Knuif, 2010, rs for odour.Attached background material Attached document Remarks 7327-60-8, appearance,Huybens,2010, CoA, REF.doc / 187.5 KB (application/msword) Applicant's summary and conclusion Conclusions 7327-60-8, NTANBased on results in this report: The substance is a off-white, odourless, organic powder (wet cake).

4.2 Melting point/freezing point Endpoint summary: Melting range UUID IUC5-1b91ae06-cf61-4282-a672-ed0285aad53d Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:10:13 CEST Remarks

Administrative Data

Short description of key information The melting range of NTAN (dried) is 125.8– 127.7°C. Discussion Key study: 7327-60-8, Melting point, Boerdijk, 2006, RS Supportedby: 7327-60-8, physchem, melting point, Witteveen, 2003, RS The melting point of the substance is 128°C. 7327-60-8, Melting point/freezing point, EPISuite, SS EPISuite calculation: 92.78°C.

Endpoint study record: 7327-60-8, Melting point, Boerdijk, 2006, RS UUID IUC5-5f9adc7b-d7b9-41f6-8ccc-3e21baa72bd4 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 00:07:57 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Stud y period May 2006 Reliability 2 (reliable with restrictions)

Rationale for reliabilit y incl. deficiencies According to OECD guidline. Data sourceReference Referenc

e type Autho

r Yea

r Title Bibliograph ic source Testing laborator y Report

no. Owner compan y Compan y study

no. Repo rt date study report Boerdij k GB 200 6 Determination of the melting point of nitrilotriacetonitr ile Akzo Nobel Technolog y & Engineerin g bv Deventer The Netherlan ds 2006059 8-DSC AkzoNob el Function al Chemical s bv 2006- 05-19

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test guideline

Qualifier Guideline Deviations according to OECD Guideline 102 (Melting point / Melting Range) no data Type of method thermal analysis GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8

EC number 230-804-9 Details on test material Batch number 2005L07241-01 (CFC-7538)Purity 94 w/w%Storage requirements Avoid contact with: aluminium, zinc, nickel, copper and copper alloys. Store in: PVC, PE, Stainless steel, bituminized tanks.Stability Stable under storage and handling conditions.Expiration date October 2008Sample preparationThe consigned sample was a wet cake. The water was removed by drying it in a desiccator filled with silica gel. The desiccator was set at vacuum at 20-25°C. After constant weight, the material is tested in the DSC. Any other information on materials and methods incl. tables

Apparatus Mettler Toledo STAR system DSC pan type Standard aluminum, 40 μl; closed Scanning rate 10.0 °C/min Weight 5 mgTemperature scanning range 25°C – 150°C Following recommendations in method: ASTM E 537-02
Results and discussions Melting / freezing point

Melt./Freez. pt. Atm. pressure Decomposition Decomp. temp. Sublimation Subl. temp. Remarks

> 125.8 — < 127.7 °C Any other information on results incl. tables Experiment Scanning rate K/min T onset °C T end-set °C T range °C

Screening DSC

Measurement 1 Measurement 2 Average value 10.0

2.0 2.0 125.8

125.8 125.9 125.8 132.3

127.6 127.8 127.7 6.5

1.8 1.9

The melting range of NTAN (dried) is 125.8 ± 0.5°C – 127.7 ± 0.5°C. Overall remarks, attachments Overall remarks Since the accuracy of the DSC sample temperature in between 25°C and at least 420°C *2 is ± 0.5°C, a melting point would be given, if T range < 1.0. The highest temperature at which the DSC is accurately calibrated. This is the melting point of Zinc(419.5°C) Above this temperature value, the accuracy decreases slowly. Attached background material Attached document RemarksCoA NTAN CFC7538.docx / 471.46 KB (application/vnd.openxmlformatsofficedocument.wordprocessingml.document) 7327-60-8, melting point, Boerdijk, 2006, REF.doc / 186 KB (application/msword) Applicant's summary and conclusion Conclusions 7327-60-8, NTAN Based on results in this report: The melting range of NTAN (dried) is 125.8– 127.7°C.

Endpoint study record: 7327-60-8, Melting point, Witteveen, 2003, RS UUID IUC5-bee3f550-c25b-481d-a18f-363b261e544e Dossier UUID 0 Author LondenK / A kzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 00:08:05 CEST Remarks

Administrative Data

Purpose flag supporting study; robust study summary Study result type experimental result Stud y period april 2003 Reliability 2 (reliable with restrictions)

Rationale for reliabilit y incl. deficiencies Similar to According to OECD guidline.Data source

Reference Referenc

e type Author Year Title Bibliographi

c source Testing laboratory Report

no. Owner company Compan y study

no. Repor t date

study report Wittevee n AJ 200 3 Thermal Analysi s of Nitrils Chemicals Research Arnhem The Netherland

s CPPM0300 9 AkzoNobe l Functional Chemicals bv 2003- 05-07

Data access data submitter is data owner Data protection claimed yes, but willing to share Cross-reference to same study 7327-60-8, physchem, boiling point,Witteveen,Thermal analysis of nitrils,rs Materials and methods Test guideline Qualifier Guideline Deviations equivalent or similar to OECD Guideline 102 (Melting point / Melting Range) Type of method thermal analysis Principles of method if other than guideline 1. TGAEquipment: TA Instruments TGA 2950Temperature program: 25°C -> 700°C at 2°C/min. Sample holders: PlatinumSample size: 6 ± 1 mg.Atmosphere: Air, 100 ml./min.2. DSCEquipment: TA Instruments DSC 2920Temperature program: -100°C ->decomposition temperature at 10°/min.Sample holders: Aluminum, standard pansSample size: 6 ± 1 mg.Atmosphere: Helium, 25 ml./min. GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8

EC number 230-804-9 Details on test material Sample code AppearanceCFC 5858 WhiteCFC 5861 WhiteNTAN batches / 5857 / 5858 are production samples, Lima, 2001/2002Since CFC samples 5143 / 5144 / 5861 contain impurities the data are not used in this data base.According to certificate of analysis of CFC-5858, 5859, 6058:Specification + typical information of NTANAnalysis of Result Method referenceAppearance Off-white wet crystals Visual, specificationAssay (NTAN) Min 90 % Calculated from water contentSpecificationWater Max 10.0 % Loss on drying (160°C) specificationChemical purity NTAN > 99% typical Results and discussionsMelting / freezing point Melt./Freez. pt. Atm. pressure Decomposition Decomp. temp. Sublimation Subl. temp. Remarks

128 °C Any other information on results incl. tables

TGA DSC RT -> 700°C, 2°C/min., air RT®decomposition, 10°C/min.,helium % H2O temp. of 0.5% wt. temp. of 1.0% wt. main

based on loss relative to loss relative to Melting peak

water-free product water-free product water-free product Onset temperature

sample Code (°C) (°C)
NTAN CFC 5858 0.0 126 125 127

NTAN CFC 5861 0.0 127 134 127

The two samples (5858 and 5861)in this data base reported show a sharp melting peak at 128°C. The other samples mentioned in the report do notshow a sharp melting peak,because of impurities. Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, melting point,Witteveen,2003, REF.doc / 187 KB (application/msword) 7327-60-8, Huybens, 2010, CoA NTAN CFC5858-5859-6058,ref.docx / 37.76 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Conclusions 7327-60-8, NTANBased on results in this report: The melting point of the substance is 128°C.

Endpoint study record: 7327-60-8, Melting point/freezing point, EPISuite, SS UUID IUC5-8e7308d9-227c-4ac3-9415-bf02a4666bd6 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 00:01:39 CEST Remarks

Administrative Data Purpose flag supporting study
Stud y result t y pe estimated by calculation Reliability 2 (reliable with restrictions) Data source Reference Reference type Author Year Title Bibliographic

source Testing laboratory Report

no. Owner company Company study no. Report date

other: EPISuiteApplicant's summary and conclusion Conclusions MP: 92.78°C

4.3 Boiling point Endpoint summary: Boiling point UUID IUC5-7090fa05-25a8-426f-925e-083a2b7e70aa Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:10:45 CEST Remarks

Administrative Data Short description of key information Melting starts at 127°C. At the same temperature the evaporation of the substance starts too. So during melting the substance evaporates, starting both at 127°C. Key value for chemical safety assessment Boilin g

point at 101 325 Pa 127 °C

Discussion Key study: 7327 -60 -8, physchem, boiling point, Witteveen, 2003, RS Supported by 7327-60-8, Bo iling point, EPISuite, SS 321.24°C Endpoint study record: 7327-60-8, Boiling point, Witteveen, 2003, RS UUID IUC5-c2dcc7db-5441-4cd5-80fb-b72fdb73bfd1 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 00:08:18 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Stud y period A pril 2003 Reliability 2 (reliable with restrictions)

Rationale for reliabilit y incl. deficiencies According to OECD guidline. Data source Reference Referenc

e type Author Year Title Bibliographi

c source Testing laboratory Report

no. Owner company Compan y study

no. Repor t date

study report Wittevee n AJ 200 3 Thermal Analysi s of Nitrils Chemicals Research Arnhem The Netherland

s CPPM0300 9 AkzoNobe l Functional Chemicals bv 2003- 05-07Data access data submitter is data owner Data protection claimed yes, but willing to share Cross-reference to same study 7327-60-8, physchem, melting point,Witteveen,Thermal analysis of nitrils,rs Materials and methods Test guideline Qualifier Guideline Deviations equivalent or similar to OECD Guideline 103 (Boiling point/boiling range) Type of method differential scanning calorimetry (and Thermogravimetric analysis) Principles of method if other than guideline Experimental conditions1. TGAEquipment: TA Instruments TGA 2950Temperature program: 25°C - >700°C at 2°C/min. Sample holders: PlatinumSample size: 6 ± 1 mg.Atmosphere: Air, 100 ml./min.2. DSCEquipment: TA Instruments DSC 2920Temperature program: -100°C ->decomposition temperature at 10°/min.Sample holders: Aluminum, standard pansSample size: 6 ± 1 mg.Atmosphere: Helium, 25 ml./min. GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8

EC number 230-804-9 Details on test material Sample code AppearanceCFC 5858 WhiteCFC 5861 WhiteNTAN batches / 5857 / 5858 are production samples, Lima, 2001/2002Since CFC samples 5143 / 5144 / 5861 contain impurities the data are not used in this data base.According to certificate of analysis of CFC-5858, 5859, 6058:Specification + typical information of NTANAnalysis of Result Method referenceAppearance Off-white wet crystals Visual, specificationAssay (NTAN) Min 90 % Calculated from water contentSpecificationWater Max 10.0 % Loss on drying (160°C) specificationChemical purity NTAN > 99% typical Results and discussions Boiling point

Boiling pt. Atm. pressure Decomposition Decomp. temp. Remarks

> 120 °C The substance evaporates during melting and before boiling.Any other information on results incl. tables

TGA DSC RT -> 700°C, 2°C/min., air RT®decomposition, 10°C/min.,helium % H2O temp. of 0.5% wt. temp. of 1.0% wt. main

based on loss relative to loss relative to Melting peak

water-free product water-free product water-free product Onset temperature

sample Code (°C) (°C)
NTAN CFC 5858 0.0 126 125 127

NTAN CFC 5861 0.0 127 134 127

Weight loss starts right after melting and th e TGA curves suggest vola tilization rather than decomposition. Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8,boiling point,Witteveen,2003, REF.doc / 191.5 KB (application/msword) 7327-60-8, Huybens, 2010, CoA NTAN CFC5858-5859-6058,ref.docx / 37.81 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Conclusions 7327-60-8, NTANBased on results in this report: melting starts at 127°C. At the same temperature the evaporation of the substance starts too.So during melting the substance evaporates, starting both at 127°C.

Endpoint study record: 7327-60-8, Boiling point, EPISuite, SS UUID IUC5-a0fab734-9cb2-4b85-bf47-f4b3899728be Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 00:01:23 CEST Remarks

Administrative Data Purpose flag supporting study
Stud y result t y pe estimated by calculation Reliability 2 (reliable with restrictions) Data source Reference Reference type Author Year Title Bibliographic

source Testing laboratory Report

no. Owner company Company study no. Report date

other: EPISuite Applicant's summary and conclusion ConclusionsBP: 321.24°C

4.4 Density Endpoint summary: Density UUID IUC5-ce70ae9a-7d59-45ee-b860-647b5f0ce85d Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 00:06:53 CEST Remarks

Administrative Data Short description of key information 7327-60-8, NTANThe relative density is 1.298 with a standard deviation of 0.001. Key value for chemical safety assessment Relative density at 20°C 1.298 Discussion Key study: 7327-60-8, Density, Knuif, 2010, RS

Endpoint study record: 7327-60-8, Density, Knuif, 2010, RS UUID IUC5-2a0bae99-d5c6-4e8c-8f7b-7e412773be12 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 00:08:37 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Stud y period A pril 2010 Reliability 1 (reliable without restriction)

Rationale for reliabilit y incl. deficiencies According to OECD 109. Data source Reference Referenc

e type Autho

r Yea

r Title Bibliograph ic source Testing laborator y Report

no. Owner compan y Compan y study

no. Repor t date

study report Knuif MJM 201 0 Determination of the relative density of nitrilotriacetonitri le (NTAN) Akzo Nobel Technolog y and Engineerin g B.V. Departme nt Powder Technolog y Deventer The Netherlan 2.416.98 4 - AkzoNob el Function al Chemical s B.V. 2010- 04-13ds

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test guideline Qualifier Guideline Deviations according to OECD Guideline 109 (Density of Liquids and Solids) (July 1995) no

according to other guideline: DIN 51913, Testing of carbon materials – Determination of density by gas pycnometer (volumetric) using helium as the measuring gas – Solid materials, May 2001 no

Type of method pycnometer method GLP compliance no (Internal Management System) Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8

EC number 230-804-9 Details on test material Batch number CFC-7538Description Off-white odourless powderPurity (NTAN) 94.1 m/m % , certificate of analysis included in the report.Water 3.2 m/m %Storage requirements Store in a cool, well ventilated area. The thermal stability depends on temperature and time of exposure. Keep container tightly closed.Expiration date April 2011 Any other information on materials and methods incl. tables Test principle

The volume of a sample of known mass is determined by measuring the changes of gas volume within the pycnometer upon introducing the sample. The volume change is obtained indirectly by measuring the changes of the pressure within the pycnometer and calculating the volume using the pressure-volume relationship for ideal gases. The volume obtained by this procedure is related to the solid alone without its gas accessible pores. The pycnometer is placed inside of a climate chamber and acclimatized at a temperature of 20.0+ 0.5oC. Measurement is carried out at this temperature.

Sample preparation

None, relative density of the sample was determined “as is”. Sample was not dried because of possible evaporation of crystal water during heating.

Test conditions Test equipmentApparatus Type Supplier Apparatus number Calibration
Analytical

balance AG 204 Mettler 2106-2A002 29.09.2009

Climate

box SLH 60/40 Weiss Enet 3122-3S001 09.06.2009

Helium pycnometer AccuPyc 1340 Gas Pycnometer Micromeritics 2109-5S001 25.03.2010

Gas Gas Type Supplier

Helium 5.0 Air Liquide

Results and discussion Density Type Densit y Temp. Remarks relative density 1.2979 20 °C St. Dev. 0.0007 RSD 0.05% Any other information on results incl. tables The true density of the NTAN was determined in triplicate. True density of NTAN

Sample Sample weight

(g) Volume (cm 3 ) True density (g/cm 3 ) Temperature (oC)

CFC-7538.1 3.5491 2.7352 1.2976 20.0 CFC-7538.2 3.3541 2.5853 1.2974 20.0 CFC-7538.3 3.3547 2.5833 1.2986 20.0

The relative density (D 4 20 ), is theratioof the truedensityof a material at a temperature of 20oC and the density of water at a temperature of 4oC (density = 0.999975g/cm 3 ). The relative density has no dimension. Relative density of NTAN:

Sample / Relative Density NTAN Batch No.: CFC-7538
Average 1.2979 St. Dev. 0.0007 RSD (%) 0.05 Overall remarks, attachments Overall remarks Control pycnometer

Control sample Standard volume

(cm 3 ) Measured volume

(cm 3 ) Deviation (%) Temperature (oC)
St.0 (empty vial) 0.0000 0.0011 - 20.1

Standard 135 6.3717 6.3719 +0.003% 20.1 Remark: The measured volume of0 should be within 0.05% of full scale (10 cm 3 ). The measured volume of the Standard 135 should be within 0.01%.Attached background material Attached document Remarks 7327-60-8, relative density, Knuif, 2010, REF.doc / 192 KB (application/msword) Applicant's summary and conclusion Conclusions 7327-60-8, NTAN Based on results in this report: The relative density is 1.298 with a standard deviation of 0.001.

4.5 Particle size distribution (Granulometry) Endpoint summary: Particle size distribution (Granulometry) UUID IUC5-c077fbb7-0815-490e-8594-c4c0a8a0cfbf Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 00:09:44 CEST Remarks

Administrative Data Short description of key information 10% of the particles is smaller than 90-120 μm, 50% of the particles is smaller than 185-250 μm and 90% of the particles is smaller than 305-370 μm. In addition, no NTAN particles smaller than 10 μ m were found. Therefore, the respirable fraction is considered to be negligible. Discussion Key study: 7327-60-8, Particle size distribution (Granulometry), Reichwein, 2010, RS

Endpoint study record: 7327-60-8, Particle size distribution (Granulometry), Reichwein, 2010, rs UUID IUC5-53337715-0d38-43f5-8ab2-589a986d6073 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 00:08:46 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Stud y period January 2005 Reliability 1 (reliable without restriction) Rationale for reliability incl. deficiencies According to NEN-ISO 13320-1. Data source Reference Referenc

e type Author Yea

r Title Bibliographi

c source Testing laboratory Report

no. Owner compan y Compan y study

no. Repor t date

study report Reichwei n AM 201 0 Particle size distributio n of AkzoNobel Chelates (FC-C), Deventer, FC-C M1002 8 Akzo Nobel Functiona l 2010- 03-10NTAN The Netherland

s Chemical

s

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Type of distribution volumetric distribution Test guideline

Qualifier Guideline Deviations according to other guideline: NEN-ISO 13320-1 no data GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material CFC 5858 (lot no. 801L023873-05)CFC 5859 (lot no. 801L023874-05) CFC 6058 (lot no. 801L024271- 05). These commercial samples produced by AkzoNobel in Lima (U.S.) were tested.The actual product is produced commercially as a wet cake with a very low dustiness Any other information on materials and methods incl. tables Laser diffraction Equipment: Sympatec HELOS with RODOS and GRADIS RODOS: dry dispersion with aerosol jet GRADIS: dry dispersion by gravity In order to be able to measure the particle size distribution by laser diffraction, NTAN was measured as such in a dry air stream (RODOS). Results and discussions Particle size Percentile Mean St. dev. Remarks D10 > 92 — < 120 μm Different samples D50 > 185 — < 246 μm Different samples D90 > 304 — < 371 μm Different samples Particle size distribution at different passages No. Size Distribution Remarks < 10 μm 0 % Any other information on results incl. tables The first graph for each sample shows a cumulative value for the particle size distribution and the second graph shows a frequency value for the particle size distribution. The key data that characterize the particle size distributions for these three NTAN samples are given in the table below.Characteristics CFC 5858 lot no. 801L023873-05 CFC 5859 lot no. 801L023874-05 CFC 6058 lot no. 801L024271-05 Q3 ( 10.00 μm ) [%] 0.0 0.0 0.0 X3 ( Q=10.00 % ) [μm] 92 100 120 X3 ( Q=50.00 % ) [μm] 185 200 246 X3 ( Q=90.00 % ) [μm] 304 332 371 Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, Particle size distribution, Reichwein, 2010, REF.doc / 190 KB (application/msword) Applicant's summary and conclusion Conclusions 7327-60-8,NTAN:10% of the particles is smaller than 90-120 μm, 50% of the particles is smaller than 185- 250 μm and 90% of the particles is smaller than 305-370 μm. In addition, no NTAN particles smaller than 10 μ m were found. Therefore, the respirable fraction is considered to be negligible.

4.6 Vapour pressure Endpoint summary: Vapour pressure UUID IUC5-8a164af6-6c5e-4432-9265-701a9ac4b48c Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:36:34 CEST Remarks

Administrative Data Short description of key information NTAN was determined to have a vapour pressure of 0.000104 Pa at 25°C. Discussion Key study: 7327-60-8, Vapour pressure, Elliott 2009, RS Supported by: 7327-60-8, Vapour pressure, Grybat, 2006, SS The vapour pressure is < 0.001 mbar at 80°C. The substance starts to decompose above 80°C. 7327-60-8 Vapour pressure, EPISuite, SS EPISuite calculation: 6.17x10 -5 mmHg

Endpoint study record: 7327-60-8, Vapour pressure, Elliott, 2009, RS UUID IUC5-d1e3904a-7f00-4e61-b32e-301e34403b7a Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 10:26:14 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Stud y period September-October 2009 Reliability 1 (reliable without restriction)Rationale for reliabilit y incl. deficiencies Well conducted and reported study under GLP Data source Reference Referenc

e type Autho

r Yea

r Title Bibliographi

c source Testing laboratory Repor t no. Owner compan y Compan y study

no. Repor t date

study report Elliott T 201 0 Nitrilotriacetonitri le (CAS# 7327- 60-8): Vapor pressure determination by the Gas Saturation Method ABC Laboratorie s Inc., Columbia, Missouri, USA 64965 Akzo Nobel Function al Chemicals LLCChelates 2010- 02-05

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test guideline

Qualifier Guideline Deviations according to OECD Guideline 104 (Vapour Pressure Curve) according to EPA OPPTS 830.7950 (Vapor Pressure) Type of method gas saturation method GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material See CoA included in the report.Batch no: CFC 9161Expiration date: June 2, 2012Purity: 99.6 +/- 1.5%Water: 0.4 +/- 0.2% Results and discussions Vapour pressure 0.000104 Pa

25 °C

Remarks SD = 4.22E-6 Pa Transition / decomposition Transition / decomposition no

Overall remarks, attachments Overall remarksGiven this vapour pressure, NTAN was calculated to have a Henry's Law Constant of 1.85E-9 in deionised water at 25 degrees C. Attached background material Attached document Remarks 7327-60-8, Vapour pressure, Elliott, 2010, ref.docx / 37.32 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Conclusions NTAN was determined to have a vapour pressure of 0.000104 Pa at 25 degrees C.

Endpoint study record: 7327-60-8, Vapour pressure, Grybat, 2006, SS UUID IUC5-9b4ebe37-f7dc-4d6b-b60e-2cc869f3f087 Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 10:41:07 CET Remarks

Administrative Data

Purpose flag supporting study Study result type experimental result Study period 2006 Reliability 1 (reliable without restriction) Rationale for reliability incl. deficiencies Fair report, according to EU Method A.4, signed certificate of analysis seperately uploaded. Data source Reference Referen ce type Author Yea

r Title Bibliograp hic source Testing laboratory Report

no. Owner compan y Compa ny study

no. Repo rt date

study report Grybat A, Wollma nn R, Fischer K 200 6 Vapour pressure determination of nitrilotriacetonit rile Laboratory for Thermophysi cal Properties (LTP GmbH), Oldenburg, Germany 2006059 8-VP AkzoNob el Function al Chemica ls bv 2006- 08-03

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test guideline

Qualifier Guideline Deviations according to EU Method A.4 (Vapour Pressure) no data Type of methodgas saturation method GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8

EC number 230-804-9 Details on test material Batch number 2005L07241-01 (CFC-7538)Product code D2765Description off-white odourless powder (wet cake)Purity 94.1 w/w%Storage requirements Avoid contact with: aluminium, zinc, nickel, copper and copper alloys. Store in: PVC, PE, Stainless steel, bituminized tanks.Stability Stable under storage and handling conditions.Expiration date October 2008 Any other information on materials and methods incl. tables To make sure that no decomposition product was found in the cryo trap, the original and the frozen sample were measured by HPLC with UV/Vis detection. For the identification of the substances at the top of the observed peak the flow was stopped and a spectrum scan was performed. The test substance is measured as such. Results and discussions Vapour pressure < 0.001 mBar

80 °C

Remarks No substance could be found in the cryo trap after several days even at the highest temperature. Transition / decomposition Transition / decomposition yes

at > 80 °C Any other information on results incl. tables Several vapour pressure measurements with the gas saturation method were performed at temperatures between 60 and 120 °C for NTAN. No substance could be found in the cryo trap after several days even at the highest temperature. The substance starts to decompose above 80°C. Overall remarks, attachments Attached background material Attached document RemarksCoA NTAN CFC7538.docx / 471.46 KB (application/vnd.openxmlformatsofficedocument.wordprocessingml.document) 7327-60-8, Vapor pressure, Grybat, 2006, ref.docx / 37.43 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Conclusions 7327-60-8, NTANBased on results in this report: The determined vapour pressure was < 0.001 mbar at 80°C. The substance starts to decompose above 80°C.Endpoint study record: 7327-60-8 Vapour pressure, EPISuite, SS UUID IUC5-5721ae7d-cbd3-43f2-b9a6-3a4a1eb98eb6 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 00:06:28 CEST Remarks

Administrative Data Purpose flag supporting study
Stud y result t y pe estimated by calculation Reliability 2 (reliable with restrictions) Data source Reference Reference type Author Year Title Bibliographic

source Testing laboratory Report

no. Owner company Company study no. Report date

other: EPISuite Applicant's summary and conclusion Conclusions VP: 6.17x10-5 mmHg

Endpoint study record: 7327-60-8, Vapour pressure, Daubert & Danner, 1989, S UUID IUC5-81f93014-e2b8-465a-ad21-936d4c1ecd03 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:17:41 CEST Remarks

Administrative Data Purpose flag disregarded study Study result type experimental result Reliability 4 (not assignable)

Rationale for reliabilit y incl. deficiencies Original report was not available. Data source Reference Reference type Author Year Title Bibliographic

source Testing laboratory Report

no. Owner company Company study no. Report date publication Daubert & Danner 1989

Applicant's summary and conclusion Conclusions VP: 3.17 x 10-7 mmHg

4.7 Partition coefficient Endpoint summary: Partition coefficient UUID IUC5-acbbe867-6765-4f36-84b8-87cb0d2b95e2 Dossier UUID 0Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:19:37 CEST Remarks

Administrative Data Short description of key information Calculation withKOWWIN: Log Kow =-1.3903, indicating that NTAN is not expected to be susceptible to bioaccumulation. Key value for chemical safety assessment

Lo g Kow ( Lo g Pow ) -1.3903

Discussion Key study: 7327-60-8, Partition coefficient, Zvinavashe, 2010, RS Software used: KOWWIN v1.67 (Sept. 2008) for log Kow A computional method has been used because the substance does not sufficiently dissolve in octanol due to its polarity and therefore high solubility in water. The calculated log Kow is less than the bioconcentration threshold (log Kow =3) indicating that NTAN is not expected to be susceptible to bioaccumulation Endpoint study record: 7327-60-8, Partition coefficient, EPISuite, Zvinavashe, 2010, RS UUID IUC5-8cda2be8-bfa7-4e71-b84a-9bd4b65ba2f9 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:32:37 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type estimated by calculation Study period June 2010 Reliability 1 (reliable without restriction) Rationale for reliability incl. deficiencies A computional method has been used because the substance does not sufficiently dissolve in octanol due to its polarity and therefore high solubility in water. Data source Reference Referenc

e type Author Yea

r Title Bibliographi

c source Testing laboratory Report

no. Owner compan y Compan y study

no. Repor t date

other: Calculatio

n Zvinavash e E 201 0 Calculated octanolwater partition coefficient (log Kow) and organic carbon A kzo Nobel T&E, Arnhem, the Netherland

s 341.17 0 Akzo Nobel Functiona l Chemical s BV 2010- 06-04partition coefficient (log Koc) Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Partition coefficient type octanol-water Test guideline Qualifier Guideline Deviations no guideline required Type of method other: calculation Principles of method if other than guideline Software used: KOWWIN v1.67 (Sept. 2008) for log Kow GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material NTANSMILES : C(#N)CN(CC(#N))CC(#N) Analytical method other: calculation Any other information on materials and methods incl. tables The calculated log Kow value represents the neutral or unionized form. It should be noted that in the ionized form, the log Kow value is probably lower. The fragment constants used for estimating the log Kow value are derived from models with compounds in the log Kow range of 0 – 5. Estimations for log Kow values outside this range are generally less accurate. Results and discussions Partition coefficient

Type Partition coefficient Temp. pH Remarks log Pow -1.3903 Calculation, so no temperature or pH mentioned. Any other information on results incl. tables Log Kow calculation

SMILES : C(#N)CN(CC(#N))CC(#N)CHEM : Acetonitrile, 2,2',2''-nitrilotrisMOL FOR: C6 H6 N4 MOL WT : 134.14
-------+-----+--------------------------------------------+---------+-------- TYPE | NUM | LOGKOW FRAGMENT DESCRIPTION | COEFF | VALUE-------+-----+--------------------------------------------+---------+-------- Frag | 3 | -CH2- [aliphatic carbon] | 0.4911 | 1.4733 Frag | 1 | -N< [aliphatic attach] |-1.8323 | -1.8323 Frag | 3 | -C#N [cyano, aliphatic attach] |-0.9218 | -2.7654 Factor| 3 | -N-CH-C#N [aminomethylcyano] correction | 0.5017 | 1.5051 Const | | Equation Constant | | 0.2290
-------+-----+--------------------------------------------+---------+-------- Log Kow = -1.3903 Overall remarks, attachments Overall remarks Despite this inaccuracy, the calculated log Kow value is in accordance with th e high water solubility (3 g/L) for this compound. The calculated log Kow is less than the bioconcentration threshold (log Kow =3) indicating that NTAN is not expected to be susceptible to bioaccumulation. Attached background material Attached document Remarks 7327-60-8, partition coefficient log Kow. calculation, Zvinavashe, 2010, REF.doc / 198 KB (application/msword) Applicant's summary and conclusion Conclusions 7327-60-8, NTANCalculation withKOWWIN: Log Kow =-1.3903, indicating that NTAN is not expected to be susceptible to bioaccumulation. Executive summary The calculated log Kow is less than the bioconcentration threshold (log Kow =3) indicating that NTAN is not expected to be susceptible to bioaccumulation.

4.8 Water solubility Endpoint summary: Water solubility UUID IUC5-504e482e-fd01-4768-801f-29c72af72602 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:37:14 CEST Remarks

Administrative Data Short description of key information The water solubility is 3.06 g/L at 20°C. Key value for chemical safety assessment Water solubilit y 3.06 g/L at the temperature of 20 °C

Discussion Key study: 7327-60-8, Water solubility, Elliott, 2009, RS Supported by: 7327-60-8, Water solubility, EPISuite, SS EPISuite calculation: 1x106 mg/L

Endpoint study record: 7327-60-8, Water solubility, Elliott, 2009, RS UUID IUC5-2cac6b88-3633-44ee-a5db-af7e61908cfe Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / NetherlandsDate 2011-07-14 21:24:31 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period 18 August, 2009 till 30 November 2009 Reliability 1 (reliable without restriction) Rationale for reliability incl. deficiencies GLP, according to OECD Guideline 105.

Data source Reference Referenc

e type Autho

r Yea

r Title Bibliographi

c source Testing laboratory Repor t no. Owner compan y Compan y study

no. Repor t date

study report Elliott T 200 9 Nifrilotriacetonitri le (CAS# 7327- 60-8): Determination of Water Solubility ABC Laboratorie s, Inc. 7200 E. ABC Lane Columbia, Missouri 65202 ABC Study No. 64964 Akzo Nobel Function al Chemicals LLCChelates 2009- 11-30

Data access data submitter is data owner Data protection claimed yes, but not willing to share Materials and methods Test guideline

Qualifier Guideline Deviations according to OECD Guideline 105 (Water Solubility) no

according to other guideline: U.S. Environmental Protection Agency Product Properties Test Guidelines OPPTS 830.7840 no

Type of method flask method GLP compliance yes (incl. certificate) Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test materialProduct Name: Nitrilotriacetonitrile (NTAN)Lot Number : CFC 9161Purity: 99.6% a.i. Expiration Date: June 02, 2012 Any other information on materials and methods incl. tables Preliminary Test Approximately 0.1 g of test substance was weighed into a graduated mixing cylinder (10 mL volume). Water was added in incremental volumes (to equal 0.1, 0.4, 1,2, and 10 nt total volume) with 10 minutes of agitation between additions. Since the sample did not dissolve after the addition of 10 mL of reagent water, the solution was quantitatively transferred to 100 -ml cylinders and brought to a volume of 100 mL with water. The test substance still did not show visual solubility, so the solution was quantitatively transferred to a 1000 -ml graduated cylinder and tested at volumes of 250, 500, and 1,000 mL. The test substance did not show visual solub ility at a volume of 1,000 mL, therefore the solubility of all solutions was estimated to be less than 0.0001 g/mL. Method Validation The analytical method was validated for recovery of the test substance in triplicate at nominal concentration levels of 41.4 and 414μg/mL. Samples were prepared by transferring 10 mL of the 2.O7-mg/mI~ nitrilotriacetonitrile solution to a 25-mL volumetric flask and bringing to volume with reagent water for a concentration of 828μg/mL. The low level (41.4- [tg/mL) method validation samples were prepared by transferring 0.5 mL of the 828 -μg/niL solution to a 10-mL volumetric flask and bringi ng to volume with acet onitrile. The high level (414 -μglmL) method validation samples were prepared by transferring 5-nt of the 828μg/mL solution to a 10-nt volumetr ic flask and bringing to volume with acetonitrile. The analytical method was validated for linearity of response by injecting six standards ranging from 20.5 to 656μg/ml(Section 2.1.2). The analytical method was validated for system precision and suitability by injecting a mid range standard (82.0 -μg/mL), as prepared above, ten consecutive times. Suitability was also shown by injecting a blank samp le to check for interference. The samples were analyzed using the GC equipment and conditions described in the report. Definitive Test Definitive test samples were prepared at three proposed sample points in reagent water by transferring approximately I g of the test substance into 7 -ml Teflon® centrifuge tubes. To each tube was added 5 mL of reagent water. These amounts allowed for an excess of test substance in all samples. A quality control sample at each of the 3 time points was also prepared by adding approximately 10 mg of the te st substance to 2-oz amber bottles. Fifty milliliters of reagent water was added to each bottle. The test and quality control samples were capped and shaken by hand to mix. They were placed on a shaker in an incubator at 30°C. After approximately 24 hours, the test and quality control samples for the first time point were removed from the shaker in the 30°C incubator and were placed on a shaker in the 20°C environmental chamber. After approximately 48 hours, the test and quality control samples for the second time point were removed from the shaker in the 30°C incubator and were placed on a shaker in the 20°C environmental chamber. After approximately 72 hours, the test and quality control samples for the third time point were removed from the shaker in the 30°C incubator and were placed on a shaker in the 20°C environmental chamber. After allowing each of the samples to remain on the shaker in the 20°C chamber for approximately 24 hours, the samples were centrifuged for 30 minutes at approximately 45,000 x g and 20°C. The supematant of each sample was transferred to a 14 -ml amber vial with a disposable pipet. The pH of each supematant was measured after transferred to the amber vial. The samples were diluted by transferring 1 mL to 25 -ml volumetric flasks, and bringing the flasks to volume with acetonitrile. The diluted sa mples were further diluted by transferring 1 mL to I O-mL volumetric flasks and bringing the flasks to volume with acetonitrile. The samples were analyzed by GC using the conditions described in the report. Results and discussions Water solubility > 3.02 — < 3.1 g/LTemp. 20 °C pH 6.9 Any other information on results incl. tables Solubility The solubility of nitrilotriacetonitrile was dete rmined to be 3,060 ± 41.1μg/ml(3.06 g/L) at 20+/-0.5°C, with less than 1% variability from the mean in each of the three replicates. A representative chromatogram of a definitive test sample is presented in the report. The pH of the supernatants were 6.90, 6.88, and 6.87. Method Validation Acceptable recoveries of nitrilotriacetonitrile were ob tained at levels similar to those seen in the water solubility samples. Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, water solubility, Elliot, 2009, RE F.doc / 191 KB (application/msword) Applicant's summary and conclusion Interpretation of results soluble (1000-10000 mg/L) Conclusions 7327-60-8, NTAN,According to this report: The water solubility is 3.06 g/l at 20°C.

Endpoint study record: 7327-60-8, Water solubility, EPISuite, SS UUID IUC5-a682fbf0-8d23-4d96-a6b5-1eb7c531b31b Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:24:12 CEST Remarks

Administrative Data Purpose flag supporting study
Stud y result t y pe estimated by calculation Reliability 2 (reliable with restrictions) Data source Reference Reference type Author Year Title Bibliographic

source Testing laboratory Report

no. Owner company Company study no. Report date

other: EPISuite Applicant's summary and conclusion Interpretation of results very soluble (> 10000 mg/L) Conclusions WS: 1x10E6 mg/l

4.10 Surface tension Endpoint summary: Surface tension UUID IUC5-9a1f8559-89fe-4d6b-bd9d-452f93424887 Dossier UUID 0Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:38:57 CEST Remarks

Administrative Data Short description of key information Based on the molecular structure the endpoint could be waived. No hydrofobic tail is present. However, measurement of the surface tension of an aqueous test substance solution at a concentration of 1.06 g/l and at a temperature of 22.0°C resulted in a value of 72.6 mN/m.This is the same surface tension as water has. Key value for chemical safety assessment Surface tension 72.6 in mN/m at 20°C and concentration in m g /L 1060

Discussion Key study: 7327 -60 -8, Surface tension, Van der Linden, 2006, RS

Endpoint study record: 7327-60-8, Surface tension, Van der Linden, 2006, RS UUID IUC5-df8bf7d3-78db-41e3-99b0-18ece4399afe Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:39:24 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Stud y period Januari 2006 Reliability 1 (reliable without restriction)

Rationale for reliabilit y incl. deficiencies According to EU A.5 and OECD 115 guidelines. Data source Reference Referenc

e type Autho

r Yea

r Title Bibliograph ic source Testing laborator y Report

no. Owner compan y Compan y study

no. Repor t date

study report Linde

n van der EGM 200 6 Determination of the surface tension of an aqueous solution of nitrilotriacetonitr ile Research and Technolo gy Chemical

s 2006059 8-ST AkzoNob el Function al Chemical s bv 2006- 01-17

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methodsType of method OECD harmonised ring method Test guideline

Qualifier Guideline Deviations according to EU Method A.5 (Surface Tension) yes (Cleaning procedure)

accordin g to OECD Guideline 115 ( Surface Tension of Aqueous Solutions ) y es ( Cleanin g procedure ) Principles of method if other than guideline Deviations from the standard procedureAll glass vessels used for the test were new. All glassware was thoroughly washed with tap-water and rinsed with Direct Q Ultrapure water. Before each measurement the measuring body was rinsed with Direct Q Ultrapure water and shortly heated above a flame. GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8

EC number 230-804-9 Details on test material Batch number :2005L07241-01 (CFC-7538)Product code :D2765Description: Off white odourless powder (wet cake)Purity: 94.1 w/w%Storage requirements: Avoid contact with: aluminium, zinc, nickel, copper and copper alloys. Store in: PVC, PE, Stainless steel, bituminized tanks.Stability : Stable under storage and handling conditions.Expiration date: October 2008 Details on methods EquipmentApparatus: Krüss Processor Tensiometer K12 (version 3.1). The measuring body is a platinum-iridium ring with a wire radius of 0.185 mm and a ring radius of 9.55 mm (ring no. 5).Water: Millipore Direct Q Ultrapure Water system (CAP-CWA lab).Rationale: The test system is recognized by international guidelines as recommended apparatus.Validation: The test system is validated before measurement using 96% ethanol ([7.3] for reference value) or n-hexane, n-heptane or n-octane ([7.4] for reference values). Any other information on materials and methods incl. tables Preparation of the test solution

An amount of 106.0 mg test substance was dissolved in 100 ml Direct Q Ultrapure water, resulting in a clear test solution. The concentration of the test solution was 1.06 g/l.

Sample measurement

A clean measurement vessel is filled wi th the test solution. The time from transferring the test solution to the measurement vessel is recorded. The measurement vessel is placed on the mobile sample table. The ambient temperature is read from a thermometer in the laboratory. The table is raised until the measurement body is just above the surface of the solution. The table automatically moves upward until the ring is immersed below the surface of the solution. Subsequently, the table is automatically lowered gradually and evenly to detach the ring from the surface until the maximum force is reached. The force is read on the tensiometer. The liquid layer attached to the ring must not separate from the ring. This measurement is repeated until a constant surface tension value is reached.

DATA interpretationThe surface tension is read from th e tensiometer in mN/m (SI-sub-unit).

In order to calculate the correct surface tension, the value read on the tensiometer is multiplied by the calibration factor Ф b . This yields a value which is only approximate and requires a correction. The additional correction is based on the Harkins-Jordan tables [7.5], see Table 1. The table provides corrected values for measurements taken after calibration. Interpolation shall be used for readings ranging between the table values.

Correction of the water calibrated surface tension according to Harkins and Jordan

Density of the test solutions: ρ ≈ 1 g / cm 3

. Average ring radius: R = 9.55 mm.. Average ring wire diameter: r = 0.185 mm.

Experimental value after water calibration (mN/m) Corrected value (mN/m)

20 18.1 22 20.1 24 22.1 26 24.1 28 26.1 30 28.1 32 30.1 34 32.1 36 34.1 38 36.1 40 38.2 42 40.3 44 42.3 46 44.4 48 46.5 Experimental value after water calibration (mN/m) Corrected value (mN/m)

50 48.6 52 50.7 54 52.8 56 54.9 58 57.0 60 59.1 62 61.3 64 63.4 66 65.5 68 67.7 70 69.9 72 72.0 74 74 76 76 78 78

Results and discussions Surface tension 72.6 mN/m Temp. 22 °C
Concentration 1.06 g/L Any other information on results incl. tables Surface tension of Direct Q Ultrapure water at 22.0°C

.

Measurement no. Surface tension (mN/m) measured mean σ g 1 71.23 71.20 2 71.19 3 71.19The calibration factor Ф b = 72.44 / 71.20 = 1.017. Surface tension measurement of test solution

The test solution was transferred to the measurement vessel within 1 hour after preparation. Table 3 shows the individual measurements and the corrected surface tension (calibration correction and Harkins & Jordan correction) for the test solution in water at a concentration of 1.06 g/l and at a temperature of 22.0°C.

Surface tension of an aqueous test substance solution at 22.0°C

measurement time expired 1 (min) Surface tension (mN/m)

measured mean 2 not corrected mean 2 after calibration correction mean after Harkins &

Jordan correction 1 3 71.37 2 9 71.42 71.38 72.59 - 3 13 71.35

1 Time passed since the test solution was transferred to the measurement vessel. 2 Mean was calculated from measurements 1 - 3.

The surface tension was found to be stable after 3 minutes.

Overall remarks, attachments Attached background material Attached document RemarksCoA NTAN CFC7538.docx / 471.46 KB (application/vnd.openxmlformatsofficedocument.wordprocessingml.document) 7327-60-8, surface tension, Van der Linden, 2006, REF.doc / 194 KB (application/msword) Applicant's summary and conclusion Conclusions 7327-60-8 NTAN, Based on results in this report:The surface tension of an aqueous test substance solution at a concentration of 1.06 g/l and at a temperature of 22.0°C is 72.6 mN/m. Based on the criteria as outlined in the EEC guideline, the substance is classified as “not surface active”.

4.11 Flash point Endpoint summary: Flash point UUID IUC5-0a2fc5c8-5c5b-4d49-bd0a-e724cc0037d3 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:39:49 CEST Remarks

Administrative Data Short description of key information The endpoint is waived because it is only a relevant property for liquids, thus this test does not need to be performed for substances that are solids or gases at room temperature. Endpoint study record: 7327-60-8, Flash point, waiver UUID IUC5-50d8504f-77f3-4883-87b5-d433c6207222Dossier UUID 0 Author ArtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:36:30 CET Remarks

Administrative Data
Data waiving other justification

Justification for data waiving Only a relevant property for liquids, thus it does not need to be done for substances that are solids or gases at room temperature.

4.12 Auto flammability Endpoint summary: Auto flammability UUID IUC5-821c296a-3d46-48c4-8341-7e6f0a988ea7 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:40:09 CEST Remarks

Administrative Data Short description of key informationThe substance did not show self-heating up to its melting point which means that it does not have a selfignition temperature. Discussion Key study: 7327-60-8, Auto flam mability, Nage lhout, 2010, RS Endpoint study record: 7327-60-8, Auto flammability, Nagelhout, 2010, RS UUID IUC5-5ecaaf7d-a7cc-44a8-9074-d6ac44cafb1d Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:40:24 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period March 25 – 26, 2010 Reliability 1 ( reliable without restriction ) Rationale for reliability incl. deficiencies Performed under Internal management system, according to EU Method A.16. Data source Reference Referenc

e type Author Yea

r Title Bibliograph ic source Testing laboratory Report

no. Owner compan y Compan y study

no. Repor t date

study report Nagelho ut G 201 0 Determinati on of the self-ignition temperature of NTAN Akzo Nobel Technology and Engineering B.V. 2.419.35 7 Akzo Nobel Function al Chemical 2010- 06-15Department HSE Safety,Testi ng & Modeling, Deventer, The Netherlands s BV

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test guideline

Qualifier Guideline Deviations according to EU Method A.16 (Relative Self-Ignition Temperature for Solids) no GLP compliance no (Internal quality management system) Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8

EC number 230-804-9 Details on test material Batch number CFC-7538Description Off-white odourless powderCertificate of analysis included in the report:Purity 94.1%Water 3.2%Storage requirements Store in a cool, well ventilated area. The thermal stability depends on temperature and time of exposure . Keep container tightly closed.Expiration date April 2011 Any other information on materials and methods incl. tables Test principle A certain amount of substance is introduced into a wire mesh cube and placed in an air circulation oven at room temperature. The temperature in the centre of the sample is recorded while the temperature of the oven is increased to 400°C, or to the melting point if lower, at a rate of 0.5°C/min. When the substance ignites the sample temperature will show a very sharp rise above the oven temperature. The temperature of the oven at which the sample temperature reaches 400°C by self-heating is called the self-ignition temperature. Test conditions

Equipment Temperature programmed air circulation oven Thermocouples Rössel Chromel-Alumel, type K; diameter 0.5 mm for sample and 1.0 mm for oven Recorder Atal MS3 Datalogger Sample cube 20 mm sides, wire mesh 0.045 mm Scan rate 0.5°C/min Temperature range 25°C – 400°C Intake 4.66 g The test cube was filled with substance, and by tapp ing it gently, more of th e substance was added untilthe cube was completely full. The cube was then placed in the centre of the oven at room temperature. One thermocouple was placed in the centre of the cube to measure the sample temperature, and the other one between the cube and the oven wall to measure the oven temperature. The oven was closed and turned on. The temperature of the sample and oven were continuously recorded while the temperature of the oven was increased at a rate of 0.5°C/min. The test was stopped after the sample had molten completely. The thermocouples and recorder were calibrated (and adjusted when necessary) maximum one year before this stud y accordin g to the internal calibration method KALTDR-001 Results and discussions Autoflammability / Self-ignition temperature

Remarks The substance did not show self-heating up to its melting point which means that it does not have a self-ignition temperature. Any other information on results incl. tables Self-ignition temperature test substance The temperature/time course of the test is shown in the graph in the report. The temperature of the sample remained constant at 126°C due to melting of the sample. No exothermal self-heating occurred before melting. The molten sample dripped out of th e cube, resulting in an empty cube after complete melting of the sample. The recorded value of the thermocouple in the test cube had no relevance anymore thereafter, since it measured the air temperature. The test was stopped at that point.

This result means that the test substance did not show self-heating up to its melting point, and consequently it does not have a self-ignition temperature. Overall remarks, attachments Overall remarks Calibration Calibration date: July 21, 2009 After correction the deviation of the thermocouples was within the estimated accuracy of the test method, which is ±1.5°C. Attached background material Attached document Remarks 7327-60-8, self-ignition_temperature, Nagelhout, 2010, REF.doc / 196 KB (application/msword) Applicant's summary and conclusion Conclusions 7327-60-8, NTAN,The substance did not show self-heating up to its melting point which means that it does not have a selfignition temperature.

4.13 Flammability Endpoint summary: Flammability UUID IUC5-235f78cd-74ba-4179-a941-d9236ff2699f Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:41:36 CEST Remarks

Administrative Data Short description of key information The substance is not flammable and has no pyrophoric properties. Key value for chemical safety assessmentFlammabilit y non flammable Discussion Key study: 7327-60-8, Flammability, Ruiter, 2002, RS Key study: 7327-60-8, Pyrophoric properties, Ruiter, 2002, RS Justification for classification or non-classification The substance is not flammable and has no pyrophoric properties. Therefore it is not classified. Endpoint study record: 7327-60-8, Flammability, Ruiter, 2002, RS UUID IUC5-b1c119dc-0896-432b-9b45-4ee9f4cb522e Dossier UUID 0 Author ArtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:39:39 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period 2002 Reliability 2 (reliable with restrictions) Rationale for reliability incl. deficiencies Fair report, no detailed information, according to EU Method A.10 , purity mentioned. Data source Reference Referenc

e type Autho

r Yea

r Title Bibliographi

c source Testing laboratory Report

no. Owner company Compan y study

no. Repor t date

study report Ruiter AJ 200 2 Physicochemical propertie s of NTAN AkzoNobel Polymer Chemicals Deventer The Netherland

s PCS 20020013 0 AkzoNobe l Functional Chemicals B.V. 2002- 03-11

Data access data submitter is data owner Data protection claimed yes, but willing to share Cross-reference to same study 7327-60-8, Explosiveness, Ruiter, 2002, rs7327-60-8, Pyrophoric properties, Ruiter, 2002, rs7327-60- 8,Oxidising properties,Ruiter, 2002, rs Materials and methods Test guideline

Qualifier Guideline Deviations according to EU Method A.10 (Flammability (Solids)) no data Principles of method if other than guideline No data GLP compliance

noTest materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Batch number : 01036N348704, manufactured by A.N. Lima, Ohio - USASample : CFC 5422Appearance : white powderPurty > 99% on dry base. Results and discussions Solid/liquid: Ignition on contact with air

no Solid: Burning time (s)

Remarks No ignition of substance and consequently no propagation of burning Aerosol: Flame extension (cm)

Remarks The substance is not produced nor transported as an aerosol. Aerosol: Flame projection (cm)

Remarks The substance is not produced nor transported as an aerosol. Gas: Lower explosion limit (%)

Remarks The substance is a solid. Gas: Upper explosion limit (%)

Remarks The substance is a solid. Any other information on results incl. tables No ignition of substance and consequently no propagation of burning. The substance is not flammable. Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, Safety, Ruiter, 2002a, REF.doc / 183.5 KB (application/msword) Applicant's summary and conclusion Interpretation of results non flammable Conclusions 7327-60-8,NTAN,The substance is not flammable.

Endpoint study record: 7327-60-8, Pyrophoric properties, Ruiter, 2002, RS UUID IUC5-e2938470-99d6-44f9-a5bc-963ddd90abb7 Dossier UUID 0 Author ArtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:39:09 CET RemarksAdministrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period 2002 Reliability 2 (reliable with restrictions) Rationale for reliability incl. deficiencies Fair report, no detailed information, according to EU Method A.10 , purity mentioned. Data source Reference Referenc

e type Autho

r Yea

r Title Bibliographi

c source Testing laboratory Report

no. Owner company Compan y study

no. Repor t date

study report Ruiter AJ 200 2 Physicochemical propertie s of NTAN AkzoNobel Polymer Chemicals Deventer The Netherland

s PCS 20020013 0 AkzoNobe l Functional Chemicals B.V. 2002- 03-11

Data access data submitter is data owner Data protection claimed yes, but willing to share Cross-reference to same study 7327-60-8, Explosiveness, Ruiter, 2002, rs7327-60-8, Flammability, Ruiter, 2002 , rs 7327-60-8,Oxidising properties,Ruiter, 2002, rs Materials and methods Test guideline

Qualifier Guideline Deviations according to EU Method A.13 (Pyrophoric Properties of Solids and Liquids) no data Principles of method if other than guideline No data GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Batch number : 01036N348704, manufactured by A.N. Lima, Ohio - USASample : CFC 5422Appearance : white powderPurty > 99% on dry base. Results and discussions Solid/liquid: Ignition on contact with airno Remarks see: 7327-60-8, Flammability, Ruiter, 2002, rs Solid: Burning time (s)

Remarks No ignition of substance and consequently no propagation of burning Aerosol: Flame extension (cm)

Remarks The substance is not produced nor transported as an aerosol. Aerosol: Flame projection (cm)

Remarks The substance is not produced nor transported as an aerosol. Gas: Lower explosion limit (%)

Remarks The substance is a solid. Gas: Upper explosion limit (%)

Remarks The substance is a solid. Any other information on results incl. tables No ignition within five minutes after contact with diatomacéous earth The substance has no pyrophoric properties. Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, Safety, Ruiter, 2002b, REF.doc / 184 KB (application/msword) Applicant's summary and conclusion Interpretation of results other: No pyrophoric properties. Conclusions 7327-60-8,NTAN,The substance has no pyrophoric properties.

4.14 Explosiveness Endpoint summary: Explosiveness UUID IUC5-f41bfe02-fd1d-436f-b476-d6f96c756741 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:42:27 CEST Remarks

Administrative Data Short description of key information The substance has no explosive properties. Key value for chemical safety assessment Explosiveness non explosive Discussion Key study: 7327-60-8, Explosiveness, Ruiter, 2002, RSJustification for classification or non-classification The substance has no explosive properties. Therefore the substance is not classified. Endpoint study record: 7327-60-8, Explosiveness, Ruiter, 2002, RS UUID IUC5-38e4c978-bd66-4204-97c7-0d1677eaa969 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:42:46 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Stud y period 2001-2002 Reliability 2 (reliable with restrictions)

Rationale for reliabilit y incl. deficiencies According to EU Method A.14. Data source Reference Referenc

e type Autho

r Yea

r Title Bibliographi

c source Testing laboratory Report

no. Owner company Compan y study

no. Repor t date

study report Ruiter AJ 200 2 Physicochemical propertie s of NTAN AkzoNobel Polymer Chemicals Deventer The Netherland

s PCS 20020013 0 AkzoNobe l Functional Chemicals B.V 2002- 03-11

Data access data submitter is data owner Data protection claimed yes, but willing to share Cross-reference to same study 7327-60-8, Flammability, Ruiter, 2002, rs 7327-60-8, Pyrophoric properties, Ruiter, 2002, rs7327-60- 8,Oxidising properties,Ruiter, 2002, rs Materials and methods Test guideline

Qualifier Guideline Deviations according to EU Method A.14 (Explosive properties) no data Principles of method if other than guideline No data GLP compliance

no Test materials Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test materialBatch number : 01036N348704, manufactured by A.N. Lima, Ohio - USASample : CFC 5422Appearance : white powderPurty > 99% on dry base. Any other information on materials and methods incl. tables Method :EC test A.14, part thermal sensitivity, Koenen test Sample mass : 26 ml filling in steel tube Method : EC test A.14, part mechanical sensitivity (shock), BAM fallhammer apparatus Fall height : 0.4 m Drop weight : 10 kg Method : EC test A.14, part mechanical sensitivity, BAM friction apparatus Load : 360 N Results and discussions Explosive (not specified)

no Any other information on results incl. tables Thermal sensitivity, Koenen test Orifice diameter(mm) Result

2 no explosion (3 times) 6 no explosion (3 times) Result : No explosion in the tests with an orifice of 2 and 6 mm diameter. Conclusion : The substance is not thermally sensitive (limiting diameter < 2.0 mm). Mechanical sensitivity(shock) : Result: 6 times negative, shock sensitivity is > 40 J. Conclusion : The substance is not sensitive to mechanical shock Mechanical sensitivity, BAM friction apparatus Result : No external changes or decomposition effects, friction sensitivity > 360 N Conclusion : The substance is not sensitive to mechanical friction. Overall remarks, attachments Overall remarks The substance has no explosive properties. Attached background material Attached document Remarks 7327-60-8, Safety, Ruiter, 2002c, REF.doc / 187.5 KB (application/msword) Applicant's summary and conclusion Interpretation of results non explosive Conclusions 7327-60-8, NTAN,The substance has no explosive properties.

4.15 Oxidising properties Endpoint summary: Oxidising properties UUID IUC5-fb83e887-50d8-4f19-814d-8111257475cc Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:44:07 CEST Remarks

Administrative Data Short description of key informationThe substance has no oxidising properties. Key value for chemical safety assessment Oxidisin g no Discussion Key study: 7327-60-8, Oxidisin g properties, Ru iter, 2002, RS Justification for classification or non-classification The substance has no oxidising properties. Therefore it is not classified.

Endpoint study record: 7327-60-8,Oxidising properties,Ruiter, 2002, RS

UUID IUC5-c9181310-5a4f-4209-92b8-7efa3522070c Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:43:22 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Stud y period 2002 Reliability 1 (reliable without restriction)

Rationale for reliabilit y incl. deficiencies According to EU Method A.17. Data source Reference Referenc

e type Autho

r Yea

r Title Bibliographi

c source Testing laboratory Report

no. Owner company Compan y study

no. Repor t date

study report Ruiter AJ 200 2 Physicochemical propertie s of NTAN AkzoNobel Polymer Chemicals Deventer The Netherland

s PCS 20020013 0 AkzoNobe l Functional Chemicals B.V. 2002- 03-11

Data access data submitter is data owner Data protection claimed yes, but willing to share Cross-reference to same study 7327-60-8, Explosiveness, Ruiter, 2002, rs7327-60- 8, Flammability, Ruiter , 2002, rs 7327-60-8, Pyrophoric properties, Ruiter, 2002, rs Materials and methods Test guideline

Qualifier Guideline Deviations according to EU Method A.17 (Oxidising Properties (Solids)) no data Principles of method if other than guideline No dataGLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Batch number : 01036N348704, manufactured by A.N. Lima, Ohio - USASample : CFC 5422Appearance : white powderPurty > 99% on dry base. Contact with powdered cellulose Duration of test (contact time) > 147 — < 607 s Details on methods Reference material: BariumnitrateThe test was repeated with NTAN mixed with Kieselguhr (inert material). Results and discussions Test result other: The substance has no oxidising properties. Any other information on results incl. tables Test with cellulose and as reference Barium nitrate. The sample mixed with cellulose in various ratios burned faster than the reference material, Bariumnitrate. Because of the chemical nature of NTAN oxidizing properties were not to be expected. According to test procedure as prescribed in test A17 to demonstrate whether a “false” positive test result was observed, only due to burning properties of NTAN on carrier, the test was repeated with NTAN mixed with Kieselguhr (inert material). In case propagation of burning is also observed with Kiezelguhr, the propagation is only due to burning properties instead of oxidizing properties. Test with Kiezelguhr and as reference Barium nitrate. As expected, Bariumnitrate did not propagate burning. NTAN did show propagation with inert Kiezelguhr. Therefore, the propagation as determined in the tests with NTAN and cellulose mixture is due to burning properties of the substance on carrier rather than due to oxidizing properties. Even under nitrogen atmosphere, NTAN mixed with cellulose showed no propagation or smouldering effect after ignition, which demonstrates that air is needed and thus ordinary burning occurs during the original test. Overall remarks, attachments Overall remarks Conclusion : The substance has no oxidising properties. Attached background material Attached document Remarks 7327-60-8, Safety, Ruiter, 2002d, REF.doc / 189 KB (application/msword) Applicant's summary and conclusion Interpretation of results no oxidising properties Conclusions 7327-60-8, NTANThe substance has no oxidising properties.

4.17 Stability in organic solvents and identity of relevant degradation productsEndpoint summary: Stability in organic solvents and identity of relevant degradation products UUID IUC5-6524c674-276f-4cac-9195-4a7b66242201 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:43:36 CEST Remarks

Administrative Data Short description of key information Endpoint is waived. Based on the partition coefficient it can be concluded that the substance is extremely low soluble in organic solven ts and consequently the stability in organ ic solvent is considered not critical. Endpoint study record: 7327-60-8, Stability in organic solvents and identity of relevant degradation products, waiver UUID IUC5-352bf287-eab1-4951-8395-6e2009756ef9 Dossier UUID 0 Author ArtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:42:32 CET Remarks

Administrative Data
Data waiving other justification

Justification for data waiving Based on the partition coefficient it can be concluded that the substance is extremely low soluble in organic solvents and consequently the stability in organic solvent is considered not critical.

4.21 Dissociation constant Endpoint summary: Dissociation constant UUID IUC5-326150aa-c2ae-48cb-ac7b-9fe1e23ac71b Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:44:57 CEST Remarks

Administrative Data Short description of key information Endpoint is waived, study technically not feasible, no dissociation possible. Discussion Based on structure: Substance cannot dissociate due to a lack of ionisable groups. Endpoint study record: 7327-60-8, Dissociation constant, waiver UUID IUC5-c3ac1cab-4e0b-4f63-a54c-c0e69ea81d96 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:43:13 CET Remarks

Administrative Data
Data waiving study technically not feasibleJustification for data waivin g Substance cannot dissociate due to a lack of ionisable groups.

4.22 Viscosity Endpoint summary: Viscosity UUID IUC5-fd28d626-f2fc-4d7d-992f-2093c9c2fffe Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:45:12 CEST Remarks

Administrative Data Short description of key information The endpoint is waived , study technically no t feasible, study is no t relevant for solids. Endpoint study record: 7327-60-8, Viscosity, waiver UUID IUC5-177b78b3-2db1-47fa-b04a-b0ebdc3762bb Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:43:41 CET Remarks

Administrative Data
Data waiving study technically not feasible

Justification for data waivin g Property is only relevant to liquids. Substance is a solid.

4.23 Additional physico-chemical information Endpoint summary: Self-reactivity and self-heating UUID IUC5-155f9b15-3371-4e90-bea8-889507651718 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:45:31 CEST Remarks

Administrative Data Short description of key information The following endpoints are waived: Selfreactivity and selfheating. Discussion Self reactivity: testing by DSC; no classification when decomposition energy <300 J/g or when SADT > 75°C Self heating melting point <160°C (mp = 125.8 -127.7°C) See:7327-60-8, Melting range. Endpoint study record: 7327-60-8, Self-reactivity, waiver UUID IUC5-b3bfc011-bd88-4ba7-980a-4655b581287c Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:44:24 CET Remarks

Administrative DataData waiving other justification

Justification for data waiving testing by DSC; no classification when decomposition energy < 300 J/g or when SADT > 75°C Endpoint study record: 7327-60-8, Self-heating, waiver UUID IUC5-6c79e6a0-91a5-40b7-b274-c8bb7faf7f79 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:44:54 CET Remarks

Administrative Data
Data waiving other justification

Justification for data waiving melting point < 160°C (mp = 125.8 -127.7°C)See: 7327-60-8, Melting range



### 5 Environmental fate and pathways Endpoint summary: Environmental fate and pathways UUID IUC5-b59a89e2-1885-4491-9217-cb0ee536b1c3 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-11 16:26:28 CET Remarks

Administrative Data Discussion The substance is registered as TII and therefore an environmental fate assessment is not required. However, with regard to degradation the following can be concluded: Nitrilotriacetonitrile is not biodegraded in the prolon ged Closed Bottle test. Ni trilotriacetonitrile should therefore not be classified as readily biodegradable. The lack of biodegradation in the Closed Bottle test does not mean that nitrilotriacetonitrile is persistent in nature because the stringency of the test procedures could account for the recalcitrance in the Closed Bottle test. With regard to mobility the fo llowing can be conlcuded: The estimated log Koc values are less than the threshold value of 3, indicating no adsorbing potential for this compound.

5.2 Biodegradation Endpoint summary: Biodegradation UUID IUC5-f44e5659-6de6-4912-aba2-dde275df3421 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:51:30 CET Remarks

Administrative Data

Discussion Nitrilotriacetonitrile is not biodegraded in the prolon ged Closed Bottle test. Ni trilotriacetonitrile should therefore not be classified as readily biodegradable. The lack of biodegradation in the Closed Bottle test does not mean that nitrilotriacetonitrileis persistent in nature because the stringency of the test procedures could account for the recalcitrance in the Closed Bottle test.

5.2.1 Biodegradation in water: screening tests Endpoint study record: 7327-60-8, Biodegradation in water: screening tests, Ginkel van, 2010, RS UUID IUC5-0bea0e5d-fb20-41f7-a9a2-548b99139980 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:48:00 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period 08-04-2010 - 08-06-2010 Reliability 1 (reliable without restriction) Rationale for reliability incl. deficiencies Test performed under GLP according guidelines with (minor) acceptable deviations, meeting all validity criteria Data source Reference Referen ce type Autho

r Yea

r Title Bibliograph ic source Testing laboratory Report

no. Owner compan y Compan y study

no. Repo rt date study report Ginkel

van CG 201 0 Biodegradability of Nitrilotriacetonitr ile (CAS 7327- 60-8) in the closed bottle test Technology & Engineering Environment al Chemistry 2.420.08 1 AkzoNob el Function al Chemical s BV T10009 C 2010- 07-01

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test type ready biodegradability Test guideline

Qualifier Guideline Deviations according to OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test) yes (acceptable deviations) Principles of method if other than guideline One minor deviation from the guidelines of the Closed Bottle test was introduced; a) ammonium chloride was omitted from the medium to prevent oxygen consumption due to nitrification (omission doesnot result in nitrogen limitation as shown by the biodegradation of the reference compound). GLP compliance yes (incl. certificate) Test materialsIdentity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8

EC number 230-804-9 Details on test material A sample of nitrilotriacetonitrile was received from AkzoNobel Functional Chemicals on 08-03-2010. The following test substance data were submitted by th e sponsor, who accepted full responsibility for the validity thereof.- chemical name (see annex 2) nitrilotriacetonitrile- CAS reg. No. 7327-60-8- purity (see annex 2) 99.6 +/- 1.5- Batch/lot no. CFC 9161- appea rance white crystals- stability not relevant in test; stable under storage conditions- solubility in water so luble- storage at room te mperature in the darkThe concentrations cited in this report refer to the as-received sample of nitrilotriacetonitrile. Study design Oxygen conditions aerobic Inoculum or test system activated sludge, domestic, non-adapted Details on inoculum Secondary activated sludge (02-04-2010) was obtained from the wastewater treatment plant Nieuwgraaf in Duiven, The Netherlands. This plant is an activated sludge plant treating predominantly domestic wastewater. The activated sludge was preconditioned to reduce the endogenous respirationrates. To this end, 400 mg Dry Weight (DW)/L of activated sludge was aerated for one week. The sludge was diluted in the BOD bottles (van Ginkel and Stroo, 1992). Duration of test (contact time) 60 d Initial test substance concentration Initial conc. Based on 4 mg/L test mat. Parameter followed for biodegradation estimation O2 consumption Details on analytical methods The dissolved oxygen concentrations were determined electrochemically using an oxygen electrode (WTW Trioxmatic EO 200) and meter(WTW OXI 530) (Retsch, Ochten, The Netherlands). The pH was measured using a Knick 765 calimatic pH meter (Elektronische Me ssgerate GmbH, Berlin, Germany). The temperature was measured and recorded with a sensor connected to a data logger. Details on study design Test bottlesThe test was performed in 0.30 L BOD (biological oxygen demand) bottles with glass stoppers.Nutrients and stock solutionsDeionized water used in the Closed Bottle test contained per liter of water with 8.5 mg KH2PO4, 21.75 mg K2HPO4, 33.3 mg Na2HPO4.2H2O, 22.5 mg MgSO4.7H2O, 27.5 mg CaCl2, 0.25 mg FeCl3.6H2O. Ammonium chloride was omitted from the medium to prevent nitrification. Sodium acetate and the test substance were added to the bottles using stock solutions of 1.0 g/L. Test proceduresThe Closed Bottle test was performed according to the study plan. The study plan was developed from ISO Test Guidelines (1994). Use was made of 10 bottles containing only inoculum, 10 bottles containing inoculum and 16.0 mg/L humic acid, 10 bottles containing inoculum, test substance andhumic acid (16.0 mg/L), and 6 bottles containing sodium acetate and inoculum. The concentrations of the test substance and sodium acetate in the bottles were 1.0 and 6.7 mg/L, respectively. Each of the prepared solutions was dispensed into the respective group of BOD bottles so that all bottles were completely filled without air bubbles. The zero time bottles were immediately analyzed for dissolvedoxygen using an oxygen electrode. The remaining bottles were closed and incubated in the dark. The bottles with the test substance and the bottles with humic acid were placed onmagnetic stirrer plates (600 rpm). The bottles contained magnetic bars in the bottles. Two duplicate bottles of all series were withdrawn for analyses of the dissolved oxygen concentration at day 7, 14, 21, and 28. One extension from the protocol of the Closed Bottle test was introduced. The Closed Bottle test was prolonged by measuring the course of the oxygen decrease in the bottles of day 28 using a special funnel. This funnel fitted exactly in the BOD bottle. Subsequently, the oxygen electrode was inserted in the BOD bottle to measure the oxygen concentration. The medium dissipated by the electrode was collected in the funnel. After withdrawal of the oxygen electrode the medium collected flowed back into the BOD bottle, followed by removal of the funnel and closing of the BOD bottle (van Ginkel and Stroo 1992)Calculation of the resultsCalculation of endogenous respiration The endogenous respiration (oxygen depletion in the control) was calculated as follows;Oxygen depletion (endogenous respiration) (mg/L) = Mc (day 0) - Mc (day 28) Mc is the mean oxygen level in the control bottle inoculated with activated sludge.Calculation of the theoretical oxygen demand (ThOD)The ThODs of nitrilotriacetonitrile,and sodium acetate were calculated from their molecular formulae and molecular weights The calculated theoretical oxygen demand of nitrilotriacetonitrile, is 1.1 mg/mg. The theoretical oxygen demand of sodium acetate is 0.8 mg/mg. Calculation of the biochemical oxygen demand (BOD)Provided that the oxygen concentrations in all bottles at the start of the test were equal, the amounts of oxygen consumed in test and reference compound bottles were calculated as follows:Oxygen consumption (mg/L) by test substance = Mch - Mt Oxygen consumption (mg/L) by reference compound = Mc - Ma Mc or ch = the mean oxygen level in the control bottles inoculated with activated sludge n days after the start of the test (C). The control of the test substance contained humic acid (CH).Mt or a = the mean oxygen concentration in the bottles containing the test compound (t) or the reference compound, sodium acetate (a), and inoculated with activated sludge n-days after the start of the test. The biological oxygen demand (BOD) mg/mg of the test compound and sodium acetate was calculated by dividing the oxygen consumption by theconcentration of the test substance and sodium acetate in the closed bottle, respectively.Calculation of the biodegradation percentagesThe biodegradation was calculated as the ratio of the biochemical oxygen demand (BOD) to the theoretical oxygen demand (ThOD). Reference substance acetic acid, sodium salt Any other information on materials and methods incl. tables Sodium acetate anhydrous was used as a reference substance in the Closed Bottle test. This compound was purchased from Acros organics. The substance data were submitted by the supplier. · Reference compound acetic acid, sodium salt · CAS reg. No. 127-09-3 · purity >99% · batch/lot No. A0206783001 · appearance white crystals · solubility in water soluble in water · storage at ambient temperature in the dark Concentration sodium acetate in test = 6,7 mg/l Results and discussions % Degradation of test substance %De g r. St. dev. Parameter Samplin g time Remarks 0 O2 consumption 28 d 0 O2 consumption 60 d Details on results ToxicityInhibition of the degradation of a well-degra dable compound, e.g. sodium acetate by the test compound in the Closed Bottle test was not determined because possible toxicity of nitrilotriacetonitrile to microorganisms degrading acetate is not relevant. A slight inhibition of the endogenousrespiration of the inoculum by the test substance was detected. The test substance may therefore be slightly inhibitory tothe inoculum.Test conditionsThe pH of the media was 7.0 at the start of the test. The pH of the medium at day 28 was 7.0 (control and test). Temperatures were within the prescribed temperature range of 22 to 24°C Overall remarks, attachments Overall remarks Oxygen consumption (mg/L) and the percentages biodegradation of nit rilotriacetonitrile (BOD/ThOD) and sodium acetate (BOD/ThOD) in the Closed Bottle test. Time (days) Oxygen consumption (mg/L) Biodegradation (%) Test substance Acetate Test substance Acetate 0 0.0 0.0 0 0 7 0.0 3.6 0 67 14 0.0 4.0 0 74 21 0.0 0 28 0.0 0 42 0.0 0 60 0.0 0 Attached background material Attached document Remarks 7327-60-8, biodegradation in water screening tests, van Ginkel, 2010, REF.doc / 194.5 KB (application/msword) Illustration (picture/graph)

Applicant's summary and conclusion Validity criteria fulfilled yes (endogenous respiration of <1.5 mg/L; differences between replicates at day 28 < 20%; sodiumacetate was degraded 74% after 14 days.; oxygen concentrations >0.5 mg/L in all bottles during the test period.) Interpretation of results inherently biodegradable Conclusions Test performed under GLP according guidelines with (minor) acceptable deviations, meeting all validity criteriaNitrilotriacetonitrile is not biodegraded in the prolonged Closed Bottle test. Nitrilotriacetonitrile should therefore not be classified as readilybiodegradable. The lack of biodegradation in the Closed Bottle test does not mean that nitrilotriacetonitrile is pe rsistent in nature because thestringency of the test procedures could account for the recalcitrance in the Closed Bottle test. Executive summary In order to assess the biotic degradation, a ready biodegradability test was performed which allows the biodegradability to be measured in an aerobic aqueous medium. The ready biodegradability was determined in the Closed Bottle test performed according to slightly modified OECD, EU and ISO Test Guidelines, and in compliance with the OECD principles of Good Laboratory Practice. Nitrilotriacetonitrile did cause a slight reduction in the endogenous respiration. The test substance may therefore be slightly inhibitory to the inoculum. Nitrilot riacetonitrile was not biodegraded in the Closed Bottle test after 28 and 60 days (prolonged) and should therefore not be classified as readily biodegradable. The lack of biodegradation in the Closed Bottle test does not mean that nitrilotriacetonitrile is persistent in nature because the stringency of the test procedures could account for the recalcitrance in the Closed Bottle test. The test is valid as shown by an endogenous respiration of 0.9 mg/L and by the total mineralization of the reference compound, sodium acetate. Sodium acetate was degraded 74% of its theoretical oxygen demand after 14 days. Finally, the most important criterion was met by oxygen concentrations >0.5 mg/L in all bottles during the test period.

5.4 Transport and distribution 5.4.1 Adsorption / desorption Endpoint summary: 7327-60-8, Adsorption / desorption UUID IUC5-a8f3ffa8-6244-43c7-bd29-19fd5e7a43ad Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:47:55 CEST Remarks

Administrative Data Short description of key information Calculation with KOWWINv2.00 : Log Koc=1.15 (Estimate from MC) or 0.87 L/kg (Estimate from Log Kow.). The estimated log Koc values are less than the threshold value of 3, indicating no adsorbing potential for this compound. Key value for chemical safety assessment

Koc

at 20 °C 1.15

Discussion Key study: 7327-60-8, Adsorption / desorption, Zvinavashe,2010, RS Software used: KOCWIN v2.00(Jan. 2009) for log Koc A computional method has been used because the substance does not sufficiently adsorb onto organic material due to its pola rity and therefore high solubility in water. Endpoint study record: 7327-60-8, Adsorption / desorption, Zvinavashe, 2010, RS UUID IUC5-b4eac447-5a2e-4830-b937-03af0b1b5c68 Dossier UUID 0Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 15:48:44 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type estimated by calculation Study period June 2010 Reliability 1 (reliable without restriction) Rationale for reliability incl. deficiencies A computional method has been used because the substance does not sufficiently adsorb onto organic material due to its polarity and therefore high solubility in water. Data source Reference Referenc

e type Author Yea

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no. Owner compan y Compan y study

no. Repor t date

other: calculatio

n Zvinavash e E Calculated octanolwater partition coefficient (log Kow) and organic carbon partition coefficient (log Koc) Akzo Nobel T&E, Arnhem, the Netherland

s 341.17 0 Akzo Nobel Functiona l Chemical s BV 2010- 06-04

Data access data submitter is data owner Data protection claimed yes, but willing to share Cross-reference to same study 7327-60-8, Partition coefficient, Zvinavashe, 2010, rs Materials and methods Study type adsorption/desorption Media other: not relevant Type of method other: calculation Test guideline Qualifier Guideline Deviations no guideline required Principles of method if other than guideline Software used: KOCWIN v2.00(Jan. 2009) for log KocGLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test materialCompound name: NTANSMILES : C(#N)CN(CC(#N))CC(#N)CHEM : Acetonitrile, 2,2',2''-nitrilotrisDetails on properties of test surrogate or analogue material Not relevant Study design Test temperature Not relevant Batch equilibrium or other method Computational methods KOCWIN v2.00(Jan. 2009) for log Koc Results and discussions Other adsorption coefficients Type Value Remarks other: log Koc 1.15 L/kg Estimate from MCI other: Log Koc 0.87 L/kg Estimate from Log Kow Results: Batch equilibrium or other method Any other information on results incl. tables Log Koc calculation

SMILES : C(#N)CN(CC(#N))CC(#N)CHEM : Acetonitrile, 2,2',2''-nitrilotrisMOL FOR: C6 H6 N4 MOL WT : 134.14 --------------------------- KOCWIN v2.00 Results ---------------------------

Koc Estimate from MCI:

--------------------- First Order Molecular Connectivity Index ........... : 4.846 Non-Corrected Log Koc (0.5213 MCI + 0.60) .......... : 3.1261 Fragment Correction(s): 3 Nitrogen to Carbon (aliphatic) (-N-C).. : -0.6382 2 Nitrile/Cyanide (-C#N) ............... : -1.3354 Corrected Log Koc .................................. : 1.1525

Estimated Koc: 14.21 L/kg <=========== Estimated log Koc: 1.15 L/kg <===========
Koc Estimate from Log Kow:

------------------------- Log Kow (Kowwin estimate) ......................... : -1.39 Non-Corrected Log Koc (0.55313 logKow + 0.9251) .... : 0.1562 Fragment Correction(s):3 Nitrogen to Carbon (aliphatic) (-N-C).. : -0.0654 2 Nitrile/Cyanide (-C#N) ............... : 0.7843 Corrected Log Koc .................................. : 0.8752

Estimated Koc: 7.503 L/kg <=========== Estimated log Koc: 0.87 L/kg <===========

Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, adsorption-desorption,log Koc. calculation, Zvinavashe, 2010, REF.doc / 195 KB (application/msword) Applicant's summary and conclusion Conclusions 7327-60-8, NTANCalculation with KOWWINv2.00 : Log Koc=1.15 (Estimate from MC) or 0.87 L/kg (Estimate from Log Kow.) Executive summary The estimated log Koc values are less than the threshold value of 3, indicating no adsorbing potential for this compound.



### 6 Ecotoxicological Information Endpoint summary: Ecotoxicological Information UUID IUC5-c38bd173-7dc3-4e3a-b9de-c79faaa28691 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-11 16:27:08 CET Remarks

Administrative Data PNEC oral Discussion As no Chemical Safety Assessment/Report is required for the transported isolated intermediate NTAN, no PNEC values have been calculated. Conclusion on classification Environmental classification justification Acute toxicity tests for algae and Daphnia magna are available for NTAN. Both tests show EC50 values far above the threshold for classification of 100 mg/l. Therefore classification as hazardous for the environment (according to Directive 67/548/EEC and Regulation 1272/2008) is not required for NTAN based on the available data. 6.1 Aquatic toxicity Endpoint summary: Aquatic toxicity UUID IUC5-8f1dd59f-e7b1-490f-8602-09085ed8b73b Dossier UUID 0 Author KolckM / Akzo Nobel Industrial Chemicals GmbH / Ibbenbüren / Germany Date 2010-10-27 14:20:32 CEST Remarks

Administrative Data DiscussionThe available studies for the aquatic toxicity of NTAN are as follows:

Guideline Test species Endpoint Result Rem a OECD 202 Daphnia magna 48h-EC50 48h-NOEC >1000 mg/l ≥ 1000 mg/l Key s GLP, OECD 201 Pseudokirchneriella subcapitata 72h-EC50 72h-NOEC >1000 mg/l 171 mg/l Key s GLP,

6.1.1 Short-term toxicity to fish Endpoint summary: Short-term toxicity to fish UUID IUC5-7f9c956b-d9d5-40b0-917d-0dcc84b386af Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:50:48 CEST Remarks

Administrative Data

Short description of key information EC50 5460 mg/L calculated with ECOSAR. Endpoint study record: 7327-60-8, Short-term toxicity to fish, ECOSAR, SS UUID IUC5-54c6dd26-e6a4-41d6-a5a8-c827ec293ab2 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:50:16 CEST Remarks

Administrative Data Purpose flag supporting study
Stud y result t y pe estimated by calculation Reliability 2 (reliable with restrictions) Data source Reference Reference type Author Year Title Bibliographic

source Testing laboratory Report

no. Owner company Company study no. Report date

other: ECOSAR Applicant's summary and conclusion Conclusions EC50: 5460 mg/L

6.1.3 Short-term toxicity to aquatic invertebrates Endpoint summary: Short-term toxicity to aquatic invertebrates UUID IUC5-ede95e58-b9ee-4c6a-b014-6e261061ec52 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:56:00 CEST RemarksAdministrative Data Short description of key information 48h-EC50 >1000 mg/l, 48h-NOEC >= 1000 mg/l for Daphnia magna (OECD TG 202) Discussion The acute toxicity of NTAN to Daphnia magna (water flea) was tested under static conditions for 48 hours in accordance with OECD TG 202 under GLP. This test, with a reliability of 1, showed no effects up to the highest concentration of 1000 mg/l. The 48h-EC50 is therefore >1000 mg/l, the 48h-NOEC is >= 1000 mg/l. A value was also calculated with ECOSAR. The calculated EC50 value is 260 mg/L. This value turns out to be too conservative. Endpoint study record: 7327-60-8, Short-term toxicity to aquatic invertebrates, Gyimesi, 2010, RS UUID IUC5-9b617e45-2e35-4b2d-a901-bb55189001e4 Dossier UUID 0 Author KolckM / Akzo Nobel Industrial Chemicals GmbH / Ibbenbüren / Germany Date 2010-10-27 14:20:33 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period 2010-04-20 until 2010-05-28 Reliability 1 (reliable without restriction) Rationale for reliability incl. deficiencies Study was well conducted according to standard guideline, with chemical analysis and GLP. Data source Reference Referenc

e type Author Yea

r Title Bibliographi

c source Testing laboratory Report

no. Owner compan y Compan y study

no. Repor t date

study report Gyimes i M 201 0 ACUTE TOXICIT Y OF NTAN TO WATER FLEAS, DAPHNIA MAGNA Akzo Nobel T&E, dept. RALS, Arnhem, the Netherland

s 2.419.89 1 Akzo Nobel Functiona l Chemical s BV 2010- 06-07

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test guideline

Qualifier Guideline Deviations according to OECD Guideline 202 (Daphn ia sp. Acute Immobilisation Test) no GLP complianceyes (incl. certificate) Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Name: NTANChemical name: Nitrilotr iacetonitrilePurity / active ingredient: 99.6 ± 1.5 %CAS reg. no.: 7327-60-8Appearance: White crystalline powderWater solubility: 0.268 wt%Lot number: CFC 9161Stability: Stable under storage and handling conditionsStorage until required: At room temperatureSupplier: Akzo Nobel Functional Chemicals - ChelatesExpiry date: June 2, 2012 Analytical monitoring
yes Details on sampling At the start of the test, samples of 10 mL were taken from all test concentrations and the control solution. These samples were taken from the additional replicate made for chemical analysis. For the samples at the end of the test, 2.5 mL of each test vessel with the same test concentration was pipetted into a sample bottle. All samples taken at the start of the test were stored in the refrigerator prior to chemical analysis. Details on analytical methods Gas Chromatography with FIDThe following conditions were found to be suitable for the determination of the test substance in the concentration range of 0- 1.2 g/L.Column: DB 17HT, 30m x 0.32mm; 0.15 μ m dFFlame gas: 70 kPa air, 100 kPa H2Carrier gas: 70 kPa HeOven: Starting temperature: 100°CWait time: 2 min.Rate temperature increase: 10°C / min.Ending temperature: 300°CWait time: 2 min.Detector: Built in Flame Ionization DetectorTemperature detector: 300°CTemperature injector: 275 °CInjection volume: 0.5 μ lSplit flow: 15 ml/ min.Integration software: Chromeleon version 6.80 RS7 Build 2528For the standards a stock solution was made in test medium. From this solution dilutions were made in the concentration range 0 – 1.2 g/L for the calibration standards. (n >= 5). Control standards from the middle range of the calibration series were analyzed at a minimum rate of one per ten samples or at least after each test series, to check the stability of the analytical system.Samples taken at the start of the test were stored at ± 4 oC in the dark until analysis. Three separate stability samples were stored under the same conditions as the test samples to check the storage st ability, by comparing the concentration of the stored stability samples to the fresh stability samples prepared on the day of analysis. Stability samples should be within 10% of the expected values. The control standards analyzed in the middle and at the end of the sampling series were within 10% of th e value calculated with the calibration curve.RepeatabilityRepeatability samples for chemical analysis were not measured. The control standards measured several times during the analys es show that repeatability was good. All control standards did not differ more than 10%. Therefore, this omission did not have an impact on the outcome of the test.LOD/LOQThe Limit of Detection (LOD) was calculated using the signal to noise ratio (S/N). The LOD is calculated as 3 * N whereas the Limit of Quantification (LOQ) is calculated as 10 * N. S/N was determined with the software (i.e. Chromeleon) based on peak height. (noise region; retention time 9.0 – 9.4 min.) The calibration standard 75 mg/L had a signal to noise ratio of 143.2 resulting in a LOD of 1.57 mg/L and a LOQ of 5.24 mg/L.System stabilityThe stability samples that were stored under the same conditions as the test samples differed less than 10% from the freshly prepared samples, therefore storage conditions were considered to be appropriate and no corrections were made for loss.LinearityThe analytical method was found to be linear over the range of 75 - 1200 mg/L of standard solutions of test substance using the conditions described above The GC calibration analyses gave a linear regression with a squared regression coefficient r2 >= 0.998. Vehicle

noDetails on test solutions The test medium, so called Dutch Standard Water (DSW), was used, having a pH of approximately 8.2, and a conductivity between 550 and 650 μs/cm, containing per liter of deionized water: 200 mg of CaCl2·2H2O, 180 mg of MgSO4·7H2O, 100 mg of NaHCO3 and 20 mg of KHCO3.The test medium was aerated before being used in the test. The air was water-saturated and purified by an activated charcoal and cotton filter.A stock solution of 1.0000 g/L of NTAN was prepared. An accurately measured amount of 0.5000 g of test material, weighed out on an analytical balance, was loaded in approximately 480 mL test medium. Because NTAN did not dissolve easily the solution was sonicated three times 15 minutes. After this the solution was clear and it was decanted in a vo lumetric flask of 500 mL and filled up to the mark. The pH of the stock solution was checked and found to be 8.0, therefore no adjustment was necessary. A clear solution was obtained.The stock solution was diluted to obtain the test solutions. The stock solution was diluted with freshly prepared test medium.A control containing only test medium was included in the test. Test organisms Test organisms (species) Daphnia magna Details on test organismsThe test was performed with Daphnia magna Clone 5 (Origin: Dr .U. Noack-Laboratorien Käthe-PaulusStr. 1, D-31157 Sarstedt, Germany) cultured in conformity with the relevant SOP. The animals used in the test were less than 24 hours old at the beginning of the test and were obtained from parent animals having an age of 2-4 weeks. Study design Test type static Water media type freshwater Limit test

no Total exposure duration 48 h Test conditions Hardness 12.5 °dH61.2 mg/L Ca and 16.9 mg/L Mg. Test temperature 20.95 - 21.25 °C pH 7.7 - 8.0 Dissolved oxygen 8.7 - 9.0 mg/l Nominal and measured concentrations 95, 171, 309, 556 and 1000 mg/L.The concentrations measured were within 80 to 120% of the nominals, therefore nominal concentrations were used to calculate the effects. Details on test conditionsTEST SYSTEM- Test vessel: 50 mL glass beakers- Type (delete if not applicable): open- Aeration: noNo. of organisms per vessel: 5- No. of vessels per concentration (replicates): 4- No. of vessels per control(replicates): 4TEST MEDIUM / WATER PARAMETERS- Source/preparation of dilution water: DSWConductivity: 614 μs/cm- Culture medium different from test medium: Yes, culture medium is Elendt M4 mediumOTHER TEST CONDITIONS- Adjustment of pH: No- Photoperiod: 16 h of ambient lightEFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilityTESTCONCENTRATIONS- Spacing factor for test concentrations: 1.8- Range finding study- Test concentrations: control, 1, 10, 100 and 1000 mg/L. - Results used to determine the conditions for the definitive study: No effects were found, therefore it was decided that a limit test at 1000 mg/L would be sufficient. In the first definitive test, several animals died in the test concentration. Therefore, a second range finding test was performed in which again no effects were found up to 1000 mg/L. Reference substance (positive control) yes (potassium dichromate) Results and discussions Effect concentrations Duration Endpoint Effect

conc. Nominal/Measured Conc. based

on Basis for effect Remarks (e.g. 95% CL)

48 h EC50 > 1000 mg/L nominal test mat. mobility

48 h NOEC >= 1000 mg/L nominal test mat. mobility

Details on results At 48 h one animal in 1000 mg/L was lying on the bottom of the test vessel. It started to swim after gentle agitation, therefore it was not considered immobile. Results with reference substance EC50 of the reference substance was found between 4.7 and 12.3 mg/L. Reported statistics and error estimates No statistics were performed because no immobile animals were seen during the test. Any other information on results incl. tables Measured concentration and recovery values of the test samples

Sample Measured conc. t=0 (mg/L) % recovery Measured conc. t=48 (mg/L) % recovery

Control < LOD - < LOD - 95 mg/L 98.9 104.1 97.8 103.0 171 mg/L 181.8 106.3 170.3 99.6 309 mg/L 297.1 96.1 299.7 97.0 556 mg/L 516.8 93.0 581.3 104.6 1000 mg/L 944.9 94.5 946.4 94.6 Overall remarks, attachments Attached full study report Attached full study report 7327-60-8, Short-term toxicity to aquatic invertebrates, Gyimesi, 2010, REF.docx / 40.73 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Validity criteria fulfilled
yes Conclusions The test substance did not give any effect up to the highest test concentration of 1000 mg/l and is therefore not toxic to Daphnia magna. Executive summary The acute toxicity of NTAN to Daphnia magna (water flea) was tested under static conditions for 48 hours in accordance with OECD, EEC guidelines for testin g chemicals. The test was performed in compliance with Good Laboratory Practice (GLP). NTAN is soluble in water. During the preliminary test, performed as a non-GLP test, 20 animals perconcentration, divided over 4 replicates, were exposed to control, 1, 10, 100 and 1000 mg/L. No effects were found, therefore it was decided that a limit test at 1000 mg/L would be sufficient. In the first definitive test, several animals died in the test concentration. Therefore, a second range finding test was performed in which again no effects were found up to 1000 mg/L. Still it was decide d to perform a full definitive test. A definitive test with nominal test concentrations in the range of 95, 171, 309, 566 and 1000 mg/L including a control was performed. The results show that the test substance had no effect up to the highest test concentration. Based on the test results the EC50 is greater than 1000 mg/L. The results of the chemical analyses show that the observed test substance concentrations during the test differed less than 20% from the nominal test concentrations. As all concentrations measured are close to the nominals the effect data are based on the nominal test concentrations.

Endpoint study record: 7327-60-8, Short-term toxicity to aquatic invertebrates, ECOSAR, SS UUID IUC5-6688ce49-2187-4da6-8020-c946c1e8a6a2 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:52:08 CEST Remarks

Administrative Data Purpose flag supporting study
Stud y result t y pe estimated by calculation Reliability 2 (reliable with restrictions) Data source Reference Reference type Author Year Title Bibliographic

source Testing laboratory Report

no. Owner company Company study no. Report date

other: ECOSAR Applicant's summary and conclusion Conclusions EC50 260 mg/L

6.1.5 Toxicity to aquatic algae and cyanobacteria Endpoint summary: Toxicity to aquatic algae and cyanobacteria UUID IUC5-f68f3b6c-0402-4292-9d01-b360de223f1b Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:55:42 CEST Remarks

Administrative Data Short description of key information 72h-ErC50 >1000 mg/l, 72h-NOEC 171 mg/l for P. subcapitata (OECD TG 201) Key value for chemical safety assessment

EC10/LC10

or NOEC for freshwater 171 mg/Lal g ae Discussion The toxicity of NTAN to freshwater algae ( P. subcapitata ) was assessed in an algal growth inhibition test in accordance with OECD TG 201 under GLP. Slight modifications to the guideline were applied to ensure good growth and pH control of the cultures. No effect was found on growth rate up to 171 mg/L, therefore the 72h-NOEC is 171 mg/L. The 72h-ErC10 is 652 mg/L. 50% effect was not reached, therefore the 72h-ErC50 is > 1000 mg/L. A value was also calculated with ECOSAR. The calculated EC50 value is 152 mg/L and the ChV is 15 mg/L. Endpoint study record: 7327-60-8, Toxicity to aquatic algae and cyanobacteria, Gyimesi, 2010, RS UUID IUC5-74de4fcb-f4ba-4e85-8021-31b9b18b2449 Dossier UUID 0 Author KolckM / Akzo Nobel Industrial Chemicals GmbH / Ibbenbüren / Germany Date 2010-10-27 14:20:34 CEST Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period 2010-04-19 until 2010-05-28 Reliability 1 (reliable without restriction) Rationale for reliability incl. deficiencies Study was well conducted according to standard guideline, with chemical analysis and GLP. Data source Reference Referen ce type Autho

r Yea

r Title Bibliograp hic source Testing laborator y Report

no. Owner compan y Compa ny study

no. Repo rt date

study report Gyime si, M. 201 0 EFFECTS OF NTAN ON THE GROWTH OF THE FRESHWATER GREEN ALGA PSEUDOKIRCHNERI ELLA SUBCAPITATA Akzo Nobel T&E, dept. RALS, Arnhem, the Netherlan ds 2.419.8 92 Akzo Nobel Chemic als BV. 2010- 06-07

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test guideline Qualifier Guideline Deviations according to OECD Guideline 201 (Alga, Growth Inhibition Test) yes (The NaHCO3 concentration of the test medium was 150 mg/L instead of 50 mg/L, as recommended by the OECD/EEC Guidelines, in order to maintain a more constant pH during the test.) GLP complianceyes (incl. certificate) Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Details on test material Name: NTANChemical name: Nitrilotr iacetonitrilePurity / active ingredient: 99.6 ± 1.5 %CAS reg. no.: 7327-60-8Appearance: White crystalline powderWater solubility: 0.268 wt%Lot number: CFC 9161Stability: Stable under storage and handling conditionsStorage until required: At room temperatureSupplier: Akzo Nobel Functional Chemicals - ChelatesExpiry date: June 2, 2012 Analytical monitoring
yes Details on sampling At the start of the test, samples of 10 mL were taken from all test concentrations and the control solution. These samples were taken from the additional replicate made for chemical analysis. For the samples at the end of the test, 3.33 mL of each test vessel with the same test concentration was pipetted into a sample bottle. For the control the same was done, but only 1.66 mL was taken from each test vessel. The samples were filtered using an Acrodisk GHP 0.45 μ m filter. All samples taken at the start of the test were stored in the refrigerator prior to chemical analysis. Details on analytical methods Gas Chromatography with FIDThe following conditions were found to be suitable for the determination of the test substance in the concentration range of 0- 1.2 g/L.Column: DB 17HT, 30m x 0.32mm; 0.15 μ m dFFlame gas: 70 kPa air, 100 kPa H2Carrier gas: 70 kPa HeOven: Starting temperature: 100°CWait time: 2 min.Rate temperature increase: 10°C / min.Ending temperature: 300°CWait time: 2 min.Detector: Built in Flame Ionization DetectorTemperature detector: 300°CTemperature injector: 275 °CInjection volume: 0.5 μ lSplit flow: 15 ml/ min.Integration software: Chromeleon version 6.80 RS7 Build 2528For the standards a stock solution was made in test medium. From this solution dilutions were made in the concentration range 0 – 1.2 g/L for the calibration standards. (n >= 5).Control standards from the middle range of the calibration series were analyzed at a minimum rate of one per ten samples or at least after each test series, to check the stability of the analytical system.Samples taken at the start of the test were stored at ± 4 oC in the dark until analysis. Three separate stability samples were stored under the same conditions as the test samples to check the storage st ability, by comparing the concentration of the stored stability samples to the fresh stability samples prepared on the day of analysis. Stability samples should be within 10% of the expected values. The control standards analyzed in the middle and at the end of the sampling series were within 10% of th e value calculated with the calibration curve.RepeatabilityRepeatability sa mples for chemical analysis we re not measured. The control standards measured several times during the analys es show that repeatability was good. All control standards did not differ more than 10%. Therefore, this omission did not have an impact on the outcome of the test.LOD/LOQThe Limit of Detection (LOD) was calculated using the signal to noise ratio (S/N). The LOD is calculated as 3 * N whereas the Limit of Quantification (LOQ) is calculated as 10 * N. S/N was determined with the software (i.e. Chromeleon) based on peak height. (noise region; retention time 9.0 – 9.4 min.) The calibration standard 75 mg/L had a signal to noise ratio of 259.5 resulting in a LOD of 0.87 mg/L and a LOQ of 2.89 mg/LSystem stabilityThe st ability samples that were stored under the same conditions as the test samples differed less than 10% from the freshly prepared samples, therefore storage conditions were considered to be appropriate and no corrections were made for loss.LinearityThe analytical method was found to be linear over the range of 75 - 1200 mg/L of standard solutions of test substance using the conditions described above The GC calibration analyses gave a linear regression with a squared regression coefficient r2 >= 0.9994. Vehicle

no Details on test solutions A stock solution of 1.0002 g/L of NTAN was prepared. An accurately measured amount of 0.5001 g of test material, weighed out on an analytical balance, was loaded in approximately 480 mL test medium.Because NTAN did not dissolve easily the solution was sonicated three times 20 minutes. After this the solution was clear and it was transferred to a volumetric flask of 500 mL and filled up to the mark. The pH of the stock solution waschecked and found to be 8.3, therefore no adjustment was necessary. A clear solution was obtained.The test solutions were prepared by addition of the adequate amounts of stock solution to the test vessels, then dilution water was added up to the required final volume. A fourth replicate was made for chemical analysis. Test organisms Test organisms (species) Pseudokirchnerella subcapitata Details on test organisms Freshwater unicellular algae P. subcapitata (CCAP 278/4) obtained from the Culture Collection of Algae and Protozoa, Institute of Freshwater Ecology,The Windermere Laboratory, Cumbria, Ambleside, United Kingdom. After purchasing this strain was cultured and maintained. Cultures on sloped agar tubes were stored at 4°C until required.The initial stock culture was inoculated with P. subcapitata from a sloped agar tube and checked for purity by microscopic means. This algal stock culture (40 mL) of P. subcapitata was regularly transferred to fresh medium to act as inoculum for testing.The absorbance of an exponentially growing stock culture was measured. The cell density was determined using the calibration curve described below. From this algal culture a dilution was prepared to obtain an initial cell density of approximately 1*10E4 cells/mL in the test medium. Study design Test type static Water media type freshwater Limit test

no Total exposure duration 72 h Test conditions Test temperature 22.1 - 22.6 °C pH 8.2 - 9.8The pH increased with 1.6 pH units. This was a little more than recommended in the guideline, but it is not considered to have an impact on the outcome of the test. Nominal and measured concentrations 95, 171, 309, 556 and 1000 mg/LThe concentrations measured were within 80 to 120% of the nominals. Only the lowest test concentration at t = 72h differed 21.5%. As the measured concentration at the start of the test was close to nominal and also all other measured test concentrations, this is not taken into account. Nominal concentrations were therefore used to calculate the effects. Details on test conditions TEST SYSTEM- Test vessel:- Initial cells density: 1* 10E4 cells/ml- Control end cells density: 1.44 * 10E6 cells/ml- No. of vessels per concentration (replicates): 3- No. of vessels per control (replicates): 6GROWTH MEDIUM- Standard medium used: yes, OECD mediumTEST MEDIUM / WATER PARAMETERS- Culture medium different from test medium: noOTHER TEST CONDITIONS- Sterile test conditions: yes- Adjustment of pH: no- Photoperiod: continuous light- Light intensity and quality: 98.6 - 99.6 μmol/m2/sEFFECT PARAMETERS MEASURED (with observation intervals if applicable) :- Determination of cell concentrations: spectrophotometerTEST CONCENTRATIONS- Spacing factor for test concentrations: 1.8- Range finding studyFrom a range finding test it was decided that a limit test at 1000 mg/L would be sufficient. In the first definitive test, no growth was found in the test concentration.A second range finding test was performed in which no effects were found up to 100 mg/L. 1000 mg/L gavea significant effect. Reference substance (positive control) yes (potassium dichromate) Results and discussions Effect concentrations Duration Endpoint Effect

conc. Nominal/Measured Conc. based

on Basis for effect Remarks (e.g. 95% CL) 72 h NOEC 171 mg/L nominal test mat. growth rate 72 h LOEC 309 mg/L nominal test mat. growth rate 72 h EC10 652 mg/L nominal test mat. growth rate

72 h EC50 > 1000 mg/L nominal test mat. growth rate

Results with reference substance (positive control) The EC50 values of the reference compound, potassium dichromate, were in the range of 0.25-2.0 mg/L. Reported statistics and error estimates Where possible, the EC10,20,50,80 values were computed from the best fitted line (leastsquares method) through the points given by the probit of the percentage of inhibition and the logarithm of the concentration of the test substance. The Lowest Observed Effect Concentration (LOEC)was determined by comparison of the growth at each concentration and the control using threshold values from the William’s test (9.5) The No Observed Effect Concentr ation (NOEC) was derived from the results as the first concentration below the LOEC value, where growth shows no significant inhibition relative to the control values. Confidence limits were computed on the basis of Fieller’s theorem if possible (9.6). All computations were performed using the TOXCALC 5.0 program. Any other information on results incl. tables Extinctions measured in the algal growth inhibition test with NTAN

Conc. Time (hours) Area Inhib. Specific Inhib. (mg/L) 0 24 48 72 % gr. rate % 0 0.007 0.027 0.193 0.795 14.400 0.067 0 0.006 0.028 0.198 0.810 14.784 0.069 0 0.006 0.028 0.208 0.851 15.516 0.070 0 0.006 0.028 0.199 0.854 15.336 0.070 0 0.006 0.025 0.187 0.822 14.592 0.070 0 0.006 0.029 0.203 0.828 15.144 0.070 Mean 0.006 0.028 0.198 0.827 14.962 0.069 0.0

95 0.006 0.028 0.199 0.803 14.724 0.070 95 0.007 0.028 0.186 0.810 14.436 0.068 95 0.006 0.026 0.175 0.763 13.620 0.069 Mean 0.006 0.027 0.187 0.792 14.260 4.7 0.069 1.1

171 0.006 0.028 0.177 0.811 14.292 0.069 171 0.006 0.026 0.181 0.818 14.424 0.069 171 0.006 0.023 0.163 0.730 12.864 0.068 Mean 0.006 0.026 0.174 0.786 13.860 7.4 0.069 1.3

309 0.007 0.026 0.144 0.697 12.024 0.065 309 0.006 0.028 0.173 0.760 13.584 0.068309 0.006 0.026 0.173 0.755 13.476 0.069 Mean 0.006 0.027 0.163 0.737 13.028 12.9 0.067 3.0

556 0.007 0.022 0.118 0.599 10.128 0.064 556 0.007 0.024 0.129 0.637 10.896 0.065 556 0.007 0.024 0.128 0.597 10.392 0.064 Mean 0.007 0.023 0.125 0.611 10.472 30.0 0.065 7.1

1000 0.007 0.017 0.069 0.325 5.544 0.056 1000 0.007 0.020 0.075 0.331 5.832 0.055 1000 0.007 0.019 0.085 0.422 7.140 0.059 Mean 0.007 0.019 0.076 0.359 6.172 58.7 0.057 18.2 Average specific growth rate and coefficient of variation of average specific growth rates during the whole test period and themean coefficient of variation for section-by-section specific growth rates in the control cultures

Time (hours) mean SD ± CV% Mean

0-24 24-48 48-72 72 hours test CV% 0.056 0.082 0.059 0.0657 0.01412 21.48

18.47 0.064 0.082 0.059 0.0681 0.01190 17.47 0.064 0.084 0.059 0.0688 0.01306 18.97 0.064 0.082 0.061 0.0689 0.01126 16.36 0.059 0.084 0.062 0.0683 0.01348 19.72 0.066 0.081 0.059 0.0684 0.01151 16.82 mean 0.0681 stdev 0.0012 CV% 1.7226 Overall remarks, attachments Attached full study report Attached full study report 7327-60-8, Toxicity to aquatic algae and cyanobacteria, Gyimesi, 2010, REF.docx / 40.69 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Validity criteria fulfilled
yes Conclusions The NOEC and ErC10 are greater than 100 mg/l and the ErC50 is greater than 1000 mg/l, therefore NTAN can be considered as not toxic to freshwater green algae. Executive summary In order to predict effects of chemicals in an aqua tic environment, the toxicity to freshwater algae was assessed. The algal toxicity was determined in the Algal Growth Inhibition Test in accordance with OECD, EEC and ISO test guidelines and with the OECD Principles of Good Laboratory Practice. Slight modifications to the guideline were applied to ensure good growth and pH control of the cultures. The toxicity of NTAN to an exponentially growing culture of P. subcapitata was determined over an exposure period of 72 hours. No effect was found on growth rate up to 171 mg/L, therefore NOEC is 171 mg/L and LOEC is 309 mg/L. The ErC10 is 652 mg/L. 50% effect was not reached, this gives an ErC50 > 1000 mg/L. The test was conducted in a minera l salts medium in acclimatized illuminated orbital incubator. Themaximum variation in pH was 1.6 units. The test is valid as shown by: - the increase of the absorbance of the control over 72 h by a factor of 144 - the mean coefficient of variation for section-by-section specific growth rates in the control cultures is 18.5%. - the coefficient of variation of average specific growth rates during the whole test period in the replicate control cultures is 1.7%. The results of the chemical analyses show that the concentrations in the test solutions at the start and the end of the test are close to the initial nominal concentration. The measured concentration stayed within 80 to 120% of the nominal. Only the lowest test concentration at t = 72h differed 21.5%. As the measured concentration at the start of the test was close to nominal and also all other measured test concentrations, this is not taken into account. Nominal concentrations are used to calculate the effects.

Endpoint study record: 7327-60-8, Toxicity to aquatic algae and cyanobacteria, ECOSAR, SS UUID IUC5-bc6c214b-0d43-481c-9842-3d3d3a4e0c39 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 21:54:42 CEST Remarks

Administrative Data Purpose flag supporting study
Stud y result t y pe estimated by calculation Reliability 2 (reliable with restrictions) Data source Reference Reference type Author Year Title Bibliographic

source Testing laboratory Report

no. Owner company Company study no. Report date

other: ECOSAR Applicant's summary and conclusion Conclusions Algae EC50: 152 mg/LAlgae ChV: 15 mg/L



### 7 Toxicological information Endpoint summary: Toxicological information UUID IUC5-68e89f10-3569-4c31-9aaf-0bebf98c0a9b Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-12 11:04:05 CET Remarks

Administrative Data Workers Long-term exposure - local effects Discussion As no Chemical Safety Assessment Report is required for the transported isolated intermediate NTAN, no DNEL values have been calculated for the worker. Because of the oral LD50 value of 50 -200 mg/kg bw; the test substance NTAN needs classification in GHS cat. III and R25 according to DSD-DPD. Bas ed on the results described, NTAN would needclassification in GHS cat. IV and R21according to DSD-DPD for acute dermal toxicity. Due to the minimal degree of skin and eye irritation observed, NTAN does not meet the criteria for classification as a skin and eye irritant. Based on the chemical structure of NTAN it is very unlikely that NTAN has skin sensitising properties. Based on the severe toxicity observed following repeated oral treatment at 45 mg/kg bw/day (death and severe clinical signs of toxicity), NTAN should be classified with R48/22 and STOT repeated exposure Cat. 2. Because both the Ames test and the chromosome aberration test showed that NTAN was not mutagenic, no classification for genotoxicity is needed. In a combined subacute/reproduction/developmental toxicity study, NTAN did not show reproduction/developmental toxicity at a level up to 45 mg/kg bw, although this level resulted in severe parental toxicity, including death. As such no classification for reproduction toxicity is warranted. General population Long-term exposure - local effects Discussion As no Chemical Safety Assessment/Report is required for the transported isolated intermediate NTAN, no DNEL values have been calculated for the general population.

7.2 Acute Toxicity Endpoint summary: Acute Toxicity UUID IUC5-21685578-d503-47e5-a5b8-df7aa0bcf4d3 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-15 15:23:58 CEST Remarks

Administrative Data Short description of key information Several acute oral toxicity studies were available, and several types of NTAN have been tested, viz. a standard batch, a recrystallized bath and a lab washed batch. In addition, a batch was tested with unknown purity. One dermal acute toxicity was available. Discussion Although there were slight differences among six acute oral toxicity studies, viz. mortality varied between 40 to 80% at 200 mg/kg bw, it was concluded that the LD50 value was ca. 200 mg/kg bw. The batch with unknown purity showed an LD50 value between ca. 50 and ca. 200 mg/kg bw. Therefore it was concluded that the LD50 was between 50 and 200 mg/kg bw, probably close to 200 mg/kg bw. The acute dermal LD50 value was > 200 mg/kg bw (highest dose tested). Three TSCA 8(e) notifications showed that the acute dermal LD50 value was (1) > 794 and < 1260 mg/kg bw, (2) > 1000 mg/kg bw, and (3) > 2000 mg/kg bw. These results indicate that NTAN is much more toxic via the oral route than via the dermal route. Justification for classification or non-classification Because of the oral LD50 value of 50 -200 mg/kg bw; the test substance NTAN needs classification in GHS cat. III and R25 according to DSD-DPD. Based on the results described above, NTAN would need classification in GHS cat. IV and R21according to DSD-DPD for acute dermal toxicity.

7.2.1 Acute toxicity: oral Endpoint study record: 7327-60-8, Acute toxicity: oral, batch 3554B, Glaza 2001a, RS UUID IUC5-1fe1cc41-41e6-4e98-ae8a-57b344fc5360 Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 10:59:36 CET RemarksAdministrative Data

Purpose flag key study Study result type experimental result Stud y period April-May 2001 Reliability 1 (reliable without restriction)

Rationale for reliabilit y incl. deficiencies Well conducted and reported study Data source Reference Referenc

e type Autho

r Yea

r Title Bibliograph ic source Testing laborator y Report

no. Owner compan y Compan y study

no. Repor t date

study report Glaza SM 200 1 Acute oral toxicity study of Nitrilotriacetonitr ile (Batch 3554B) in rats Covance Laboratori es Inc., Madison, WI, USA 1040060 0 Akzo Nobel Chemical s Inc., Madison, WI, USA 2001- 06-27

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test type acute toxic class method Limit test

no Test guideline

Qualifier Guideline Deviations according to EPA OPPTS 870.1100 (Acute Oral Toxicity) GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Batch no: 3554BDate of receipt: 3 April 2001Purity: 95 wt%Moisture: 5 wt% Test animals Species
rat Strain Sprague-DawleySex male/female Administration / exposure Route of administration oral: gavage Vehicle corn oil Details on oral exposure The test material was mixed with corn oil; 10 mg/mL for the low dose, 40 mg/mL for the high dose. Single gavage application at 5 mL/kg bw. Doses 50 and 200 mg/kg bw No. of animals per sex per dose 5 Control animals

no Details on study design Duration of observation period following administration: 14 daysFrequency of observations and weighing: several times on the day of treatment and once daily thereafterNecropsy of survivors performed: yesOther examinations performed: body weight (weekly) Statistics Not applicable Results and discussions Effect levels Sex Endpoint Effect level Based on 95% CL Remarks male/female LD50 > 50 — < 200 mg/kg bw test mat. Mortality None at 50 mg/kg bwFour males and two females (60%) at 200 mg/kg bw. Clinical signs At 50 mg/kg bw: staggered gait, wet/dark-stained urogenital area on the day of treatmentAt 200 mg/kg bw: hypoactivity, staggered gait, clonic convulsions, wet area around the mouth, red-stained face, lacrimation, wet urogenital area. In surviving animals these changes had disappeared by Day 4 (day of treatment = day 1). Body weight Surviving animals showed body weight gain throughout the study period. Gross pathology Only in animals that died (200 mg/kg bw); changes consisted of moist, red oral discharge. Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, acute toxicity, oral, batch 3554B, Glaza, 2001a, ref.docx / 38.15 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Interpretation of results Toxicity Category III Criteria used for interpretation of resultsOECD GHS Conclusions The oral LD50 was between 50 and 200 mg/kg bw (close to 200 mg/kg bw). Executive summary An acute oral toxicity study was carried out in Sprague Dawley rats. Two groups of 5 male and 5 female rats each were dosed by single gavage at 50 or 200 mg/kg bw and kept for 14 days observation. None of the rats dosed with 50 mg/kg died; 60% mortality was observed at 200 mg/kg bw. Therefore the LD50 was between 50 and 200 mg/kg bw, close to 200 mg/kg bw. Endpoint study record: 7327-60-8, Acute toxicity: oral, recrystallized, Glaza 2001b, RS UUID IUC5-a14c3607-92e9-44d3-899c-27763bdc9b65 Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 11:05:07 CET Remarks

Administrative Data

Purpose flag key study Study result type experimental result Stud y period April-May 2001 Reliability 1 (reliable without restriction)

Rationale for reliabilit y incl. deficiencies Well conducted and reported study Data source Reference Referenc

e type Autho

r Yea

r Title Bibliograph ic source Testing laborator y Report

no. Owner compan y Compan y study

no. Repor t date

study report Glaza SM 200 1 Acute oral toxicity study of Nitrilotriacetonitr ile (Recrystallized) in rats Covance Laboratori es Inc., Madison, WI, USA 1030695 3 Akzo Nobel Chemical s Inc., Madison, WI, USA 2001- 06-18

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test type acute toxic class method Limit test

no Test guideline

Qualifier Guideline Deviations according to EPA OPPTS 870.1100 (Acute Oral Toxicity) GLP compliance
yes Test materialsIdentity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Batch no: RecrystallizedDate of receipt: 3 April 2001Purity: 95 wt%Moisture: 5 wt% Test animals Species
rat Strain Sprague-Dawley Sex male/female Administration / exposure Route of administration oral: gavage Vehicle corn oil Details on oral exposure The test material was mixed with corn oil; 40 mg/mL Single gavage application at 5 mL/kg bw. Doses 200 mg/kg bw No. of animals per sex per dose 5 Control animals

no Details on study design Duration of observation period following administration: 14 daysFrequency of observations and weighing: several times on the day of treatment and once daily thereafterNecropsy of survivors performed: yesOther examinations performed: body weight (weekly) Statistics Not applicable Results and discussions Effect levels Sex Endpoint Effect level Based on 95% CL Remarks male/female LD50 ca. 200 mg/kg bw test mat. Mortality Two males and three females (50%) died within one day Clinical signs Hypoactivity, staggered gait, prostation, tremors, clonic convulsions, wet area around the mouth, flaccidity, red-stained face, dyspnoea, and wet urogenital area. In surviving animals these changes had disappeared by Day 5 (day of treatment = day 1). Body weight Surviving animals showed body weight gain throughout the study period. Gross pathologyOnly in animals that died; changes consisted of moist, red oral discharge. Overall remarks, attachments Attached background material Attached document Remarks 7326-60-8, acute toxicity, oral, recrystallized, Glaza, 2001b, ref.docx / 38.14 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Interpretation of results Toxicity Category III Criteria used for interpretation of results OECD GHS Conclusions The oral LD50 was ca. 200 mg/kg bw Executive summary An acute oral toxicity study with NTAN Recrystallized was carried out in Sprague Dawley rats. One group of 5 male and 5 female rats was dosed by single gavage at 200 mg/kg bw and kept for 14 days observation. 50% mortality was observed at 200 mg/kg bw. Therefore the LD50 was ca. 200 mg/kg bw.

Endpoint study record: 7327-60-8, Acute toxicity: oral, lab washed, Glaza 2001c, RS UUID IUC5-a81c8fd4-4fe3-44a9-8551-02c8f436801b Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-15 14:58:09 CEST Remarks

Administrative Data

Purpose flag key study Study result type experimental result Stud y period April-May 2001 Reliability 1 (reliable without restriction)

Rationale for reliabilit y incl. deficiencies Well conducted and reported study Data source Reference Referenc

e type Autho

r Yea

r Title Bibliograph ic source Testing laborator y Report

no. Owner compan y Compan y study

no. Repor t date

study report Glaza SM 200 1 Acute oral toxicity study of Nitrilotriacetonitr ile (Lab washed) in rats Covance Laboratori es Inc., Madison, WI, USA 1040059 9 Akzo Nobel Chemical s Inc., Madison, WI, USA 2001- 06-18

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methodsTest type acute toxic class method Limit test

no Test guideline

Qualifier Guideline Deviations according to EPA OPPTS 870.1100 (Acute Oral Toxicity) GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Batch no: Lab washedDate of receipt: 3 April 2001Purity: 95 wt%Moisture: 5 wt% Test animals Species
rat Strain Sprague-Dawley Sex male/female Administration / exposure Route of administration oral: gavage Vehicle corn oil Details on oral exposure The test material was mixed with corn oil; 40 mg/mL Single gavage application at 5 mL/kg bw. Doses 200 mg/kg bw No. of animals per sex per dose 5 Control animals

no Details on study design Duration of observation period following administration: 14 daysFrequency of observations and weighing: several times on the day of treatment and once daily thereafterNecropsy of survivors performed: yesOther examinations performed: body weight (weekly) Statistics Not applicable Results and discussions Effect levelsSex Endpoint Effect level Based on 95% CL Remarks male/female LD50 > 200 mg/kg bw test mat. Mortality Two males and two females (40%) were found dead on the day of treatment. Clinical signs Hypoactivity, staggered gait, tremors, clonic convulsions, wet area around the mouth, red-stained face, dyspnoea, non-formed faeces, dark-stained urogenital area. In surviving animals these changes had disappeared by Day 3 (day of treatment = day 1). Body weight Surviving animals showed body weight gain throughout the study period. Gross pathology Only in animals that died; changes consisted of moist, red oral discharge. Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, acute toxicity, oral, lab washed, Glaza, 2001c, ref.docx / 38.2 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Interpretation of results Toxicity Category III Criteria used for interpretation of results OECD GHS Conclusions Because 40% of the animals died, the oral LD50 was slightly higher than 200 mg/kg bw . Executive summary An acute oral toxicity study was carried out with NTAN Lab washed in Sprague Dawley rats. One group of 5 male and 5 female rats was dosed by single gavage at 200 mg/kg bw and kept for 14 days observation. 40% mortality was observed at 200 mg/kg bw. Therefore the LD50 was slightly higher than 200 mg/kg bw.

Endpoint study record: 7327-60-8, Acute toxicity: oral, batch 01013N34603, Glaza 2001d, RS UUID IUC5-6d9c369b-0223-4ebf-9847-f11e3f686311 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-15 15:15:27 CEST Remarks

Administrative Data

Purpose flag key study Study result type experimental result Stud y period April-May 2001 Reliability 1 (reliable without restriction)

Rationale for reliabilit y incl. deficiencies Well conducted and reported study Data source Reference Referenc Autho Yea Title Bibliograph Testing Report Owner Compan Repore type r r ic source laborator y no. compan y y study

no. t date

study report Glaza SM 200 1 Acute oral toxicity study of Nitrilotriacetonitr ile (batch 01013N34603) in rats Covance Laboratori es Inc., Madison, WI, USA 1010633 0 Akzo Nobel Chemical s Inc., Madison, WI, USA 2001- 05-04

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test type acute toxic class method Limit test

no Test guideline

Qualifier Guideline Deviations according to EPA OPPTS 870.1100 (Acute Oral Toxicity) GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Batch no: 01013N346403Purity: 94.43 wt%Moisture: 5.57 wt%White powderNTAB Test animals Species
rat Strain Sprague-Dawley Sex male/female Administration / exposure Route of administration oral: gavage Vehicle corn oil Details on oral exposure The test material was mixed with distilled water to a specific concentration fo r each doselevel (10 mg/mL for the 50 mg/kg dose level and 40 mg/mL for the 200 mg/kg doselevel). Single gavage application at 5 mL/kg bw.Doses 50 and 200 mg/kg bw No. of animals per sex per dose 5 Control animals

no Details on study design Duration of observation period following administration: 14 daysFrequency of observations and weighing: several times on the day of treatment and once daily thereafterNecropsy of survivors performed: yesOther examinations performed: body weight (weekly) Statistics Not applicable Results and discussions Effect levels Sex Endpoint Effect level Based on 95% CL Remarks male/female LD50 > 50 — < 200 mg/kg bw test mat. Mortality Five male and 3 female animals treated at 200 mg/kg were found dead on the day of test material administration. No other mortality was observed. The estimated oral LDso values in male and female rats were determined to be between 50 and 200 mg/kg of body weight. Clinical signs Clinical signs of toxicity were observed in all ten animals treated at 200 mg/kg and included hypoactivity, staggered gait, prostration, tremors, clonic convulsions, redstained face, dyspnea, wet area/foam around the mouth, few feces, and pale appearance. The two surviving females returned to a normal appearance by Day 3 after treatment. Body weight A ll animals surviving to the end of the observation period exhibited body weight gain during the study. Gross pathology Test material-related findings observed at the macroscopic necropsy examinations were limited to those animals treated at 200 mg/kg and dying on the day of test material administration. These findings included dry or moist red oral discharge. One female that also died on the day of test material administration had no test material-related findings. There were no other test material-related findings. Applicant's summary and conclusion Interpretation of results Toxicity Category III Criteria used for interpretation of results OECD GHS Conclusions None of the animals died at 50 mg/kg bw and 8/10 died at 200 mg/kg bw. The oral LD50 is therefore between 50-200 mg/kg bw. Executive summary The test material, NTAN, was evaluated for its acute oral toxicity potential in male and female rats when administered as a single gavage dose at levels of 50 and 200 mg/kg of body weight. All animals treated at 50 mg/kg appeared normal and exhibited body weight gain throughout the study. All mortality in the animals treated at 200 mg/kg (five males and three females) occurred on the day of test material administration. Clinical signs of toxicity were observed in the ten animals treated at 200 mg/kg and included hypoactivity, staggered gait, prostration, tremors, clonic convulsions, red-stained face, dyspnea, wet area/foam around the mouth, few feces, and pale appearance. The two survivingfemales returned to a normalappearance by Day 3 after treatment and exhibited body weight gain during the study. Test materialrelated findings observed at the macroscopic necropsy examinations were limited to those animals treated at 200 mg/kg and dying on the day of test material administration. These findings included dry or moist red oral discharge. There were no other test material-related findings. Based on the mortality observed in the study, the estimated oral LD50 values for male and female rats were detelmined to be between 50 and 200 mg/kg.

Endpoint study record: 7327-60-8, Acute toxicity: oral, batch 3554B, Kukulinski, 2001a, SS UUID IUC5-2ca5f8a3-9b0e-47fe-bb12-89f34943157a Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 13:42:26 CET Remarks

Administrative Data

Purpose flag supporting study Study result type experimental result Study period April-May 2001 Reliability 2 (reliable with restrictions) Rationale for reliability incl. deficiencies Limited reported study, no CoA included in the report (but most probaly similar to that in the acute oral study of Glaza (2001a). Data source Reference Referenc

e type Author Year Title Bibliographi

c source Testing laborator y Repor t no. Owner company Compan y study

no. Repor t date

study report Kukulinsk i M 200 1 Acute oral toxicity study OPPTS 870.1100, D.O.T. Design 49 CFR 173.132; Compound : NTAN Lot 3554B Tox Monitor Lab. Inc., Oak Park, IL 60302, USA 01-034 Akzo Nobel Functiona l Chemical s LLC, Dobbs Ferry NY 10522- 3401, USA 2001- 05-17

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test type acute toxic class method Limit test
yes Test guidelineQualifier Guideline Deviations according to EPA OPPTS 870.1100 (Acute Oral Toxicity) no GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Lot no: 3554B (most probably similar to that used in acute oral study by Glaza (2001a). Test animals Species
rat Strain Sprague-Dawley Sex male/female Administration / exposure Route of administration oral: gavage Vehicle corn oil Doses 200 mg/kg No. of animals per sex per dose 5 Control animals

no Any other information on materials and methods incl. tables 10% suspension in corn oil was made. This corresponds to about 100 mg/mL. Thus an animal received about 2 mL/kg bw (this was not indicated in the report). Results and discussions Effect levels Sex Endpoint Effect level Based on 95% CL Remarks male/female LD50 < 200 mg/kg bw test mat. Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, acute toxicity, oral, batch 3554B, Kukulinski 2001, ref.docx / 38.36 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Interpretation of results Toxicity Category IIICriteria used for interpretation of results OECD GHS Conclusions Because 60% died at 200 mg/kg bw, the LD50 value was slightly lower than 200 mg/kg, and as such toxicity category III OECD GHS would be required. Executive summary An acute oral toxicity study with NTAN was carried out in Sprague Dawley rats. One group of 5 male and 5 female rats was dosed by single gavage at 200 mg/kg bw and kept for 14 days observation. 60% mortality was observed at 200 mg/kg bw. Therefore the LD50 was < 200 mg/kg bw.

Endpoint study record: 7327-60-8, Acute toxicity: oral, recrystallized, Kukulinski, 2001b, SS UUID IUC5-29354066-6691-429d-a22b-dc94ea624b81 Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 13:47:42 CET Remarks

Administrative Data

Purpose flag supporting study Study result type experimental result Study period April-May 2001 Reliability 2 (reliable with restrictions) Rationale for reliability incl. deficiencies Limited reported study, no CoA included in the report (but most probaly similar to that in the acute oral study of Glaza (2001b). Data source Reference Referenc

e type Author Yea

r Title Bibliographi

c source Testing laborator y Repor t no. Owner compan y Compan y study

no. Repor t date

study report Kukulinsk i M 200 1 Acute oral toxicity study OPPTS 870.1100, D.O.T. Design 49 CFR 173.132; Compound: NTAN Recrystallize d Tox Monitor Lab. Inc., Oak Park, IL 60302, USA 01- 037 Akzo Nobel Functiona l Chemical s LLC, Dobbs Ferry NY 10522- 3401, USA 2001- 05-17

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test type acute toxic class method Limit testyes Test guideline

Qualifier Guideline Deviations according to EPA OPPTS 870.1100 (Acute Oral Toxicity) no GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Lot no: Recrystallized (mos t probably similar to that used in acute oral study by Glaza (2001b)). Test animals Species
rat Strain Sprague-Dawley Sex male/female Administration / exposure Route of administration oral: gavage Vehicle corn oil Doses 200 mg/kg No. of animals per sex per dose 5 Control animals

no Any other information on materials and methods incl. tables 10% suspension in corn oil was made. This corresponds to about 100 mg/mL. Thus an animal received about 2 mL/kg bw (this was not indicated in the report). Results and discussions Effect levels Sex Endpoint Effect level Based on 95% CL Remarks male/female LD50 > 200 mg/kg bw test mat. Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, acute toxicity, oral, recrystallized, Kukulinski 2001b, ref.docx / 38.42 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusionInterpretation of results Toxicity Category III Criteria used for interpretation of results OECD GHS Conclusions Because 40% died at 200 mg/kg bw, the LD50 value was slightly higher than 200 mg/kg, and as such toxicity category III OECD GHS would be required. Executive summary An acute oral toxicity study with NTAN Recrystallized was carried out in Sprague Dawley rats. One group of 5 male and 5 female rats was dosed by single gavage at 200 mg/kg bw and kept for 14 days observation. 40% mortality was observed at 200 mg/kg bw. Therefore the LD50 was > 200 mg/kg bw.

Endpoint study record: 7327-60-8, Acute toxicity: oral, lab washed, Kukulinski, 2001c, SS UUID IUC5-8660db17-7669-410a-8dcf-5151357a28c5 Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 13:51:40 CET Remarks

Administrative Data

Purpose flag supporting study Study result type experimental result Study period April-May 2001 Reliability 2 (reliable with restrictions) Rationale for reliability incl. deficiencies Limited reported study, no CoA included in the report (but most probaly similar to that in the acute oral study of Glaza (2001c). Data source Reference Referenc

e type Author Year Title Bibliographi

c source Testing laborator y Repor t no. Owner company Compan y study

no. Repor t date

study report Kukulinsk i M 200 1 Acute oral toxicity study OPPTS 870.1100, D.O.T. Design 49 CFR 173.132; Compound : NTAN Lab washed Tox Monitor Lab. Inc., Oak Park, IL 60302, USA 01-036 Akzo Nobel Functiona l Chemical s LLC, Dobbs Ferry NY 10522- 3401, USA 2001- 05-17

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methodsTest type acute toxic class method Limit test
yes Test guideline

Qualifier Guideline Deviations according to EPA OPPTS 870.1100 (Acute Oral Toxicity) no GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Lot no: Lab washed (most probably similar to that used in acute oral study by Glaza (2001c). Test animals Species
rat Strain Sprague-Dawley Sex male/female Administration / exposure Route of administration oral: gavage Vehicle corn oil Doses 200 mg/kg No. of animals per sex per dose
5 Control animals

no Any other information on materials and methods incl. tables 10% suspension in corn oil was made. This corresponds to about 100 mg/mL. Thus an animal received about 2 mL/kg bw (this was not indicated in the report). Results and discussions Effect levels Sex Endpoint Effect level Based on 95% CL Remarks male/female LD50 ca. 200 mg/kg bw test mat. Overall remarks, attachments Attached background material Attached document Remarks7327-60-8, acute toxicity, oral, lab washed, Kukulinski 2001c, ref.docx / 38.38 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Interpretation of results Toxicity Category III Criteria used for interpretation of results OECD GHS Conclusions Because 50% died at 200 mg/kg bw, the LD50 value was ca. 200 mg/kg, and as such toxicity category III OECD GHS would be required. Executive summary An acute oral toxicity study with NTAN Lab washed was carried out in Sprague Dawley rats. One group of 5 male and 5 female rats was dosed by single gavage at 200 mg/kg bw and kept for 14 days observation. 50% mortality was observed at 200 mg/kg bw. Therefore the LD50 was ca. 200 mg/kg bw.

Endpoint study record: 7327-60-8, Acute toxicity: oral, batch 01013N34603, Kukulinski, 2001d, SS UUID IUC5-5cec13b4-333b-4462-9b4e-07139aa0f75f Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-15 15:05:36 CEST Remarks

Administrative Data

Purpose flag supporting study Study result type experimental result Study period April 28, 2001 to May 12, 2001 Reliability 2 (reliable with restrictions) Rationale for reliability incl. deficiencies Limited reported study, no CoA included in the report (but most probaly similar to that in the acute oral study of Glaza (2001c). Data source Reference Referenc

e type Author Yea

r Title Bibliographi

c source Testing laborator y Repor t no. Owner compan y Compan y study

no. Repor t date

study report Kukulinsk i M 200 1 Acute oral toxicity study OPPTS 870.1100, D.O.T. Design 49 CFR 173.132; Compound: batch 01013N3460 3 Tox Monitor Lab. Inc., Oak Park, IL 60302, USA 01- 035 Akzo Nobel Functiona l Chemical s LLC, Dobbs Ferry NY 10522- 3401, USA 2001- 05-17

Data access data submitter is data owner Data protection claimedyes, but willing to share Materials and methods Test type acute toxic class method Limit test
yes Test guideline

Qualifier Guideline Deviations according to EPA OPPTS 870.1100 (Acute Oral Toxicity) no GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Lot no: 01013N34603white powderNTAN Test animals Species
rat Strain Sprague-Dawley Sex male/female Administration / exposure Route of administration oral: gavage Vehicle corn oil Doses 200 mg/kg No. of animals per sex per dose 5 Control animals

no Any other information on materials and methods incl. tables 10% suspension in corn oil was made. This corresponds to about 100 mg/mL. Thus an animal received about 2 mL/kg bw (this was not indicated in the report). Results and discussions Effect levels Sex Endpoint Effect level Based on 95% CL Remarks male/female LD50 ca. 200 mg/kg bw test mat. Applicant's summary and conclusionInterpretation of results Toxicity Category III Criteria used for interpretation of results OECD GHS Conclusions Because 40% died at 200 mg/kg bw, the LD50 value wa s ca. 200 mg/kg, and as such toxicity category III OECD GHS would be required. Executive summary NTAN was tested for acute oral toxicity in accordance with OPPTS Guidelines and D.O.T. Design 49 CFR 173.132. NTAN was administered by gavage at a dose of 200 mg/kg body weight to five male and five female rats. There was 40% mortality at the 200 mg/kg dose level during the 14 day observation period. The acute oral LD50 was determined to be greater than 200 mg/kg body weight.

Endpoint study record: 7327-60-8, Acute toxicity: oral, Dow, 1967, SS UUID IUC5-5bb1e390-cd09-4242-aa46-8ad68707d563 Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 13:57:19 CET Remarks

Administrative Data

Purpose flag supporting study Study result type experimental result Study period 1967 Reliability 2 (reliable with restrictions)

Rationale for reliabilit y incl. deficiencies Limited reported study, used as supporting evidence Data source Reference Referenc

e type Author Yea

r Title Bibliographi

c source Testing laborator y Repor t no. Owner compan y Compan y study

no. Repor t date

study report The Dow Chemic al Compan y 196 7 Toxicological properties and industrial handling hazards of Nitrilotriacetonitri le Biochemic al Research Laboratory , Dow, Midland, Michigan, USA 12 4198 The Dow Chemica l Compan y 1967- 11-28

Data access data submitter has Letter of Access Materials and methods Test type standard acute method Limit test

no Test guidelineQualifier Guideline Deviations no guideline followed Principles of method if other than guideline Study was carried out in 1967; no guideline available GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Test animals Species
rat Strain no data Sex female Administration / exposure Route of administration oral: gavage Vehicle corn oil Doses 2% solution: 25, 50, 100, 200, 398 mg/kg bw10% solution: 63, 126, 252, 500, 1000, 2000 mg/kg bw No. of animals per sex per dose 2 Control animals

no Results and discussions Effect levels

Sex Endpoint Effect level Based on 95% CL Remarks female LD50 > 50 — < 250 mg/kg bw test mat. see below Any other information on results incl. tables In case of the 2% solution: 0% (0/2) of the animals died at 50 and 100 mg/kg bw 50% (1/2) of the animals died at 25 mg/kg bw 100% (2/2) of the animals died at 200 and 398 mg/kg bw. This means that the LD50 value of the 2% solution is somewhere between 100 and 200 mg/kg bw. In case of the 10% solution: 50% (1/2) of the animals died at 126 and 252 mg/kg bw 100% (2/2) of the animals died at 63, 126, 500, 1000, and 2000 mg/kg bw. This means that the LD50 value of the 10% solution is somewhere between 50 and 250 mg/kg bw. Overall remarks, attachments Attached background materialAttached document Remarks7327-60-8, overview, Dow, 1967, ref.docx / 36.73 KB (application/vnd.openxmlformatsofficedocument.wordprocessingml.document) Applicant's summary and conclusion Interpretation of results Toxicity Category III Criteria used for interpretation of results OECD GHS Conclusions Because LD50 value was between ca. 50 and 250 mg/kg bw, classification in GHS Cat. III is needed.

Endpoint study record: 7327-60-8, Acute toxicity: oral, Younger, 1965, SS UUID IUC5-64f0a109-08ee-4348-8420-58dd58525b5b Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 22:50:46 CEST Remarks

Administrative Data Purpose flag supporting study Study result type experimental result Study period 1965 Reliability 4 (not assignable) Rationale for reliability incl. deficiencies Very limited report only summarizin g the results. The studies were submitted to U.S. EPA under TSCA section 8(e). Data source Reference Referen ce type Author Yea

r Title Bibliograp hic source Testing laborator y Repo rt no. Owner compa ny Compa ny study

no. Repo rt date

study report Young er, F.M. 196 5 Toxicological Investigation Of: NITRILOTRIACETONIT RILE (NTAN) Younger Laboratori

es Y-65- 179 Monsan to 1965- 12-02

Applicant's summary and conclusion Executive summary LD50 (Rats, Mixed Sex) The diluted compound, 10% in corn oil, was fed by stomach tube to Sprague Dawley strain albino male and female rats. Doses 100, 126, 158, 200 mg/kg. After the approximate Minimum Lethal Dose was determined groups of male end finale rate were fed in increasing doses at increments of 0.1 fractional leg intervals at four levels designed to blanket the toxicity range thereby supplying data for calculation of the LD50 which was done according to a modification of the method of S. J. de Beer. Observations were made for toxic symptoms and the viscera of the animals that succumbed were examined macroscopically. The oral LD50 for male and female rats was placed at 150 per kilogram with lower and upper limits of 150 to 174 milligrams per kilogram.

Endpoint study record: 7327-60-8, Acute toxicity: oral, Younger, 1966, SS UUID IUC5-c7fde2ad-aa40-4cf6-a2f1-0cc0c5e90bb2 Dossier UUID 0Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 23:13:40 CEST Remarks

Administrative Data Purpose flag supporting study Study result type experimental result Study period 1965 Reliability 4 (not assignable) Rationale for reliability incl. deficiencies Very limited report only summarizin g the results. The studies were submitted to U.S. EPA under TSCA section 8(e). Data source Reference Referen ce type Author Yea

r Title Bibliograp hic source Testing laborator y Repo rt no. Owner compa ny Compa ny study

no. Repo rt date

study report Young er, F.M. 196 6 Toxicological Investigation Of: NITRILOTRIACETONIT RILE (NTAN) Younger Laboratori

es Y-65- 216 Monsan to 1966- 01-28

Applicant's summary and conclusion Executive summary LD50 (Rats, Mixed Sex) The diluted compound, 10% in corn oil, was fed by stomach tube to Sprague Dawley strain albino male and female rats. Doses 126, 158, 200, 251 mg/kg. After the approximate Minimum Lethal Dose was determined groups of male end finale rate were fed in increasing doses at increments of 0.1 fractional leg intervals at four levels designed to blanket the toxicity range thereby supplying data for calculation of the LD50 which was done according to a modification of the method of S. J. de Beer. Observations were made for toxic symptoms and the viscera of the animals that succumbed were examined macroscopically. The oral LD50 for male and female rats was placed at 198 per kilogram with lower and upper limits of 170 to 321 milligrams per kilogram.

Endpoint study record: 7327-60-8, Acute toxicity: oral, Eastman Kodak, 1971, SS UUID IUC5-8508d0c8-762c-434c-a57a-84ec39d26354 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 23:19:33 CEST Remarks

Administrative Data Purpose flag supporting study Study result type experimental result Study period 1965 Reliability 4 (not assignable) Rationale for reliability incl. deficiencies Very limited report only summarizin g the results. The studies were submitted to U.S. EPA under TSCA section 8(e). Data source Reference Reference Author Year Title Bibliographic Testing Report Owner Company Reporttype source laboratory no. company study no. date study report Eastman Kodak 1971 Eastman Kodak Applicant's summary and conclusion Conclusions LD50 50 mg/kg in rats and 50-100 mg/kg in mice.

7.2.3 Acute toxicity: dermal Endpoint study record: 7327-60-8, Acute toxicity, dermal, Babish, 1978, RS UUID IUC5-2aac1a1e-ccc3-4e72-b390-693fc8a27a46 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 16:05:30 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period 1978 Reliability 2 (reliable with restrictions) Rationale for reliability incl. deficiencies Study appears well conducted with limited reporting of methods and results and a single dose tested only and not a limit dose. Data source Reference Reference type Author Year Title Biblio g raphic

source Testing laboratory Report

no. Owner company Compan y study no. Report date study report Babish JG 1978 Acute Dermal Toxicity Study in Rabbits Food and Drug Research Laboratories K067479- 003 The Dow Chemical Company 1978- 09-20

Data access data submitter has Letter of Access Materials and methods Test type standard acute method Limit test

no Test guideline Qualifier Guideline Deviations other guideline: 16 CFR 1500. 40 no GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identityIdentifier Identity

CAS number 7327-60-8 Details on test material purity> 99%White Cr ystalline material Test animals Species rabbit Strain other: albino Sex no data Administration / exposure Type of coverage occlusive Vehicle unchanged (no vehicle) (moistened with saline) Details on dermal exposure The animals were prepared by clipping the skin of the trunk free of hair. Approximately one-half of the animals was further prepared by making epidermal abrasions every 2 or 3 centimeters longitudinally over the area of exposure. The abrasions were sufficiently deep to penetrate the stratum corneum (horny layer of the epidermis) but not to distrub the derma; that is, not to obtain bleeding.The sleeve was slipped onto the animal which is then placed in a comfortable but immobilized posit ion in a multiple animal holder. Selected doses of liquids and solutions are introduced under the sleeve. If there is slight leakage from the sleeve, which may occur during the first few hours of exposure, it is collected and reapplied. Dosage levels are adjusted in subsequent exposures (if necessary) to enable a calculation of a dose that would be fatal to 50 percent of the animals. This can be determined from mortality ratios obtained at various doses employed. At the end of 24 hours the sleeves or screens are removed, the volume of unabsorbed material (if any) is measured, and the skin reactions are noted. The subjects are cleaned by thorough wiping, observed for gross symptoms of poisoning, and then observed for 2 weeks.In this study only one dose level was chosen. Duration of exposure 24 hours Doses 200 mg/kg bw No. of animals per sex per dose 10 Control animals not required Results and discussions Effect levels

Sex Endpoint Effect level Based on 95% CL Remarks no data LD0 200 mg/kg bw test mat. no mortalities observed Mortality No mortality observed Clinical signs Decreased activity and some nasal discharge was observed, but not indicated in how many animals or at what stage after dosing these effects started and ceased.Body weight Not assessed Gross pathology No pathology reported Applicant's summary and conclusion Conclusions A dose of 200 mg/kg was not lethal to adult albino rabbits following occlusive dermal exposure (intact and abraded skin). The LD0, therefore is 200 mg/kg bw.

Endpoint study record: 7327-60-8, Acute toxicity: dermal, Younger, 1965, SS UUID IUC5-7f12b6e5-683e-48f1-9fbb-ca936df622a8 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 22:50:58 CEST Remarks

Administrative Data Purpose flag supporting study Study result type experimental result Study period 1965 Reliability 4 (not assignable) Rationale for reliability incl. deficiencies Very limited report only summarizin g the results. The studies were submitted to U.S. EPA under TSCA section 8(e). Data source Reference Referen ce type Author Yea

r Title Bibliograp hic source Testing laborator y Repo rt no. Owner compa ny Compa ny study

no. Repo rt date

study report Young er, F.M. 196 5 Toxicological Investigation Of: NITRILOTRIACETONIT RILE (NTAN) Younger Laboratori

es Y-65- 179 Monsan to 1965- 12-02

Applicant's summary and conclusion Executive summary The diluted compound was applied in increasing doses at inernen of 0.2 fractional leg intervals to the closely clipped, intact skin of New Zealand white male and female rabbits. The treated area were covered with plastic strips and the animals placed in wooden stocks for periods up to twenty four hours, after which time they were as signed to individual cages. Observations were made for toxic symptoms and the viscera of the animals that subcomed were examined macroscopically. Tle Minimum Lethal Dose by Skin Absorption in male and female rabbits was found to be greater than 794 milligrams per kilogram and less than 1260 milligrams per kilogram. It was classed as moderately toxic by skin absorption in male and female rabbits.

Endpoint study record: 7327-60-8, Acute toxicity: dermal, Younger, 1966, SS UUID IUC5-c6077f91-3665-447e-a69f-68754b0783b1 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 23:40:18 CEST RemarksAdministrative Data Purpose flag supporting study Study result type experimental result Study period 1965 Reliability 4 (not assignable) Rationale for reliability incl. deficiencies Very limited report only summarizin g the results. The studies were submitted to U.S. EPA under TSCA section 8(e). Data source Reference Referen ce type Author Yea

r Title Bibliograp hic source Testing laborator y Repo rt no. Owner compa ny Compa ny study

no. Repo rt date

study report Young er, F.M. 196 6 Toxicological Investigation Of: NITRILOTRIACETONIT RILE (NTAN) Younger Laboratori

es Y-65- 216 Monsan to 1966- 01-28

Applicant's summary and conclusion Executive summary The diluted compound was applied in increasing doses at inernen of 0.2 fractional leg intervals to the closely clipped, intact skin of New Zealand white male and female rabbits. The treated area were covered with plastic strips and the animals placed in wooden stocks for periods up to twenty four hours, after which time they were as signed to individual cages. Observations were made for toxic symptoms and the viscera of the animals that subcomed were examined macroscopically. Tle Minimum Lethal Dose by Skin Absorption in male and female rabbits was found to be greater than 2000 milligrams per kilogram. It was classed as non-lethal by skin absorption in male and female rabbits.

Endpoint study record: 7327-60-8, Acute toxicity: dermal, Eastman Kodak, 1971, SS UUID IUC5-b022c6bc-281d-42e5-b48a-381d36993ff8 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 23:21:41 CEST Remarks

Administrative Data Purpose flag supporting study Study result type experimental result Study period 1971 Reliability 4 (not assignable) Rationale for reliability incl. deficiencies Very limited report only summarizin g the results. The studies were submitted to U.S. EPA under TSCA section 8(e). Data source Reference Reference type Author Year Title Bibliographic

source Testing laboratory Report

no. Owner company Company study no. Report date study report Eastman Kodak 1971 Eastman KodakApplicant's summary and conclusion Conclusions LD50 (guinea pig) >1000 mg/kg; No clinical signs.

7.3 Irritation / corrosion Endpoint summary: Irritation / corrosion UUID IUC5-8ad7b7f3-e018-4683-9d6e-20e4188359e4 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-15 15:26:07 CEST Remarks

Administrative Data Short description of key information One skin and one eye irritation test were available, and two supporting studies. Key value for chemical safety assessment Skin irritation / corrosion slightly irritating Eye irritation slightly irritating Discussion In the skin irritation test, 4 out of 5 rabbits showed slight erythema at the end of the 4 -h treatment period. Slight erythema was still observed in 2 out of these 4 animals 48 h after treatment. No corrosivity was seen. No further observations were made. In the eye irritation test, some conjunctival reddening was seen in all animals and slight corneal opacity in one animal. The eye changes were fully reversible within 7 days. The supporting studies showed no skin and eye irritation following application of 10% and the undiluted NTAN. Justification for classification or non-classification Due to the minimal degree of skin and eye irritation observed, this substance does not meet the criteria for classification as a skin and eye irritant.

7.3.1 Skin irritation / corrosion Endpoint study record: 7327-60-8, Skin irritation / corrosion, Babish, 1978, RS UUID IUC5-a7c19fce-3937-403f-98f9-d192c6f2268f Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 16:12:38 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Reliability 2 (reliable with restrictions) Rationale for reliability incl. deficiencies Study is reported sufficeintly to understand what was done. Predates GLP. Study only designed to assess corrosivity Data sourceReference Reference type Author Year Title Biblio g raphic

source Testing laboratory Report

no. Owner compan y Compan y stud y no. Report date study report Babish JG 1978 Corrosive Dermal Study with Rabbits Food and Drug Research Laboratories Inc. K067479- 005 The Dow Chemical Company 1978- 09-20

Data access data submitter has Letter of Access Materials and methods Type of method in vivo Test guideline

Qualifier Guideline Deviations according to other guideline: see below Principles of method if other than guideline Guideline for corrosivity assay followed: The method employed is patterned after 21 CFR 191.11 as described in 49 CFR 173.240 GLP compliance

no Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test materials Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material White Crystalline powderPurity >99% Test animals Species rabbit Strain other: albino Test system Type of coverage occlusive Preparation of test site shaved Vehicle unchanged (no vehicle) (moistened with saline) Amount/concentration applied 0.5 g Duration of treatment / exposure 4 hours Observation period48 hours Number of animals 5 Control animals

no Details on study design 5 healthy, acclimated animals were shaved (back). 0.5 g of test material was applied under a 1 inch square gauze. An impervious rubber wrap was then applied to hold the material in place. Exposure duration was 4 hours and observations were made at the end of the 4 hours and then at 48 hours. No further observations were made.Scores were assigned based on the scoring scale below. Any other information on materials and methods incl. tables Scoring system Erythema and Eschar Formation: No Erythema 0 Slight erythema 1 Well defined Erythema 2 Moderate to Severe Erythema 3 Severe Erythema 4 Slight Eschar Formation 5 Severe Eschar Formation, Ulceration 6 Results and discussions Irritation / corrosion results Irritation parameter Basis Time point Score Max.

score Reversibility Remarks

overall irritation

score mean 4 h ca. 1 6 other: see below score of 1 was seen in 4 out of 5 animals

overall irritation

score mean 48 h ca. 0.5 6 other: at 48 h 2 out of 5 animals still showed slight erythema, no further observations were made Irritant/corrosive response data In 4 out of 5 animals there was a slight erythema (grade 1 out of a possible 6) at 4 hours. At 48 hours this had dissapeared in 2 of those affected. No further observations were made.No corrosivity was observed. Applicant's summary and conclusion Interpretation of results other: not-corrosive Criteria used for interpretation of results EU Conclusions The substance was slightly irritating after a 4 hour exposure. No corrosivity was observed. Due to the minimal degree of irritation observed it is unlikely that this substance would meet the criteria for classification as a skin irritant.

Endpoint study record: 7327-60-8, Skin irritation / corrosion, Dow, 1967, SS UUID IUC5-4b85f3b6-a745-4f77-a554-131b8401d4d1 Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 14:09:26 CET RemarksAdministrative Data

Purpose flag supporting study Study result type experimental result Study period 1967 Reliability 2 (reliable with restrictions)

Rationale for reliabilit y incl. deficiencies Limited reported study, used as supporting evidence Data source Reference Referenc

e type Author Yea

r Title Bibliographi

c source Testing laborator y Repor t no. Owner compan y Compan y study

no. Repor t date

study report The Dow Chemic al Compan y 196 7 Toxicological properties and industrial handling hazards of Nitrilotriacetonitri le Biochemic al Research Laboratory , Dow, Midland, Michigan, USA 12 4198 The Dow Chemica l Compan y 1967- 11-28

Data access data submitter has Letter of Access Materials and methods Type of method in vivo Test guideline Qualifier Guideline Deviations no guideline followed Principles of method if other than guideline No guideline available at that time GLP compliance

no Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test materials Test material identity

Identifier Identity

CAS number 7327-60-8 Test animals Species rabbit Strain no data Test system Type of coverage no dataPreparation of test site other: intact and abraded Vehicle other: Dowanol DPM Duration of treatment / exposure repeated application, either 3 or 9 times Observation period not indicated Number of animals not indicated Control animals

no Results and discussions Any other information on results incl. tables undiluted, intact skin, 9 applicatio ns on belly: no irritation observed undiluted, abraded skin, 3 applications on belly: no irritation observed 10% solution, intact skin, 9 applicat ions on ear: slight exfoliation 10% solution, intact skin, 9 applicat ions on belly: slight exfoliation 10% solution, abraded skin, 3 applications on belly: slight hyperaemia and exfoliation Overall remarks, attachments Attached background material Attached document Remarks7327-60-8, overview, Dow, 1967, ref.docx / 36.61 KB (application/vnd.openxmlformatsofficedocument.wordprocessingml.document) Applicant's summary and conclusion Interpretation of results not irritating Criteria used for interpretation of results expert judgment

Endpoint study record: 7327-60-8, Skin irritation / corrosion, Younger, 1965, SS UUID IUC5-7a6b7fb5-d8a8-4e46-83b0-f0c28d82d5bc Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 22:51:10 CEST Remarks

Administrative Data Purpose flag supporting study Study result type experimental result Study period 1965 Reliability 4 (not assignable) Rationale for reliability incl. deficiencies Very limited report only summarizin g the results. The studies were submitted to U.S. EPA under TSCA section 8(e). Data source Reference Referen ce type Author Yea

r Title Bibliograp hic source Testing laborator Repo rt no. Owner compa Compa ny Repo rty ny study

no. date

study report Young er, F.M. 196 5 Toxicological Investigation Of: NITRILOTRIACETONIT RILE (NTAN) Younger Laboratori

es Y-65- 179 Monsan to 1965- 12-02

Applicant's summary and conclusion Executive summary The finely ground powder and a 10.0% solution-suspension in corn oil were applied to the clipped intact skin of albino rabbits and removed after twenty four hours. The applications were covered with plastic strips to retard evaporation and avoid concentration. Observations vere made over period of several days for irritation. The data, scored according to the method of praise, Woodard and Calvery (journal of pharm. and Exp. Therapentics, Volume. 82, December, 1944). The compound was classed as non-irritating to intact rabbit skin when applied as a finely ground powder.The compound was classed as non-irritating to intact rabbit skin when applied as a 10.0% solutionsuspession in corn oil.

7.3.2 Eye irritation Endpoint study record: 7327-60-8, Eye irritation, Babish, 1978, RS UUID IUC5-07578f64-161e-4691-aa51-523b11ee726e Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-08 16:48:03 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period 1978 Reliability 2 (reliable with restrictions) Rationale for reliability incl. deficiencies Apears to be well conducted and reported sufficiently to make an assessment of the endpoint. Prior to GLP. Data source Reference Reference type Author Year Title Biblio g raphic

source Testing laboratory Report

no. Owner compan y Compan y stud y no. Report date study report Babish JG 1978 Rabbit Eye Irritation Study with fluorescein Food and Drug Research laboratories Inc. K067479- 004 The Dow Chemical Company 1978- 09-20

Data access data submitter has Letter of Access Materials and methods Type of method in vivo Test guideline Qualifier Guideline Deviationsequivalent or similar to OECD Guideline 405 (Acute Eye Irritation / Corrosion) Principles of method if other than guideline method according to 16 CFR 1500.42. GLP compliance

no Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test materials Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material White Crystalline powderPurity >99% Test animals Species rabbit Strain New Zealand White Details on test animals and environmental conditions 6 animals between 2 and 4 kg purchased from Dutchland Lab. Inc., Denver, Pennsylvania Test system Vehicle unchanged (no vehicle) Any other information on materials and methods incl. tables The eyes of each rabbit were examined with sodium fluorescein and an ultraviolet lamp (prior to instillation of the test material) a nd only those rabbits whose eyes were free of irritation and corneal lesions were used. 50 mg of the test material was in stilled in one eye of each rabbit by gently pulling the lower lid away from the eyeball to form a cup into which the test substance was dropped. The lids were then held together for one second and the animal released. The untreated eye served as a control. The eyes were examined and the grade of ocular reaction recorded at 24, 48 and 72 hours for all animals using a scalar score (see attachment). The eyes were again examined with sodium fluorescein and an ultraviolet lamp at 72 hours after application of the test material to confirm the presence or absence of corneal lesion. Results and discussions Overall irritation / corrosion results Irritation parameter Basis Time point Score Max. score Reversibility Remarks cornea score mean average of 24, 48 and 72 0.11 4 fully reversible

iris score mean 24, 48, 72 hr mean 0 2 other: no effects conjunctivae score mean 24, 48, 72 hr mean 1.1 3 fully reversible chemosis score mean 24, 48, 72 hours mean 0 4 other: no effects Irritant/corrosive response data In all animals there was some conjunctival reddening that did not persist to 7 days. One animal had a slight corneal opacity that did not persist to 7 days. This substance is not considered irritating to the eyes.Any other information on results incl. tables Cornea Iris Conjuctiva red Animal 24 48 72mean 24 48 72mean 24 48 72mea n 1 0 1 1 0.666667 0 0 0 0 2 1 0 2000 0 0 0 0 0210 3000 0 0 0 0 0211 4000 0 0 0 0 0220 5000 0 0 0 0 0210 6000 0 0 0 0 0210

Mean 0.111111 0 Overall remarks, attachments Attached background material Attached document Remarks K-067479-004 scoring system.pdf / 76.66 KB (application/pdf) Scoring system Applicant's summary and conclusion Interpretation of results not irritating Criteria used for interpretation of results EU Conclusions The test material was not irritating to the eyes.

Endpoint study record: 7327-60-8, Eye irritation, Dow, 1967, SS UUID IUC5-f5a7e59c-91dd-416d-8069-f9d97edcf56a Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 14:26:24 CET Remarks

Administrative Data Purpose flag supporting study Study result type experimental result Study period 1967 Reliability 2 (reliable with restrictions)

Rationale for reliabilit y incl. deficiencies Limited reported study, used as supporting evidence Data source Reference Referenc

e type Author Yea

r Title Bibliographi

c source Testing laborator y Repor t no. Owner compan y Compan y study

no. Repor t date

study report The Dow Chemic al Compan y 196 7 Toxicological properties and industrial handling hazards of Nitrilotriacetonitri le Biochemic al Research Laboratory , Dow, Midland, Michigan, USA 12 4198 The Dow Chemica l Compan y 1967- 11-28Data access data submitter has Letter of Access Materials and methods Type of method in vivo Test guideline Qualifier Guideline Deviations no guideline followed Principles of method if other than guideline No guideline available at that time GLP compliance

no Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test materials Test material identity

Identifier Identity

CAS number 7327-60-8 Test animals Species rabbit Strain no data Test system Vehicle unchanged (no vehicle) Duration of treatment / exposure No information Observation period No information Number of animals No information Control animals

no Results and discussions Overall irritation / corrosion results Irritation parameter Basis Time point Score Max.

score Reversibilit y Remarks

overall irritation

score other: qualitative judgement not indicated no irritation observed Any other information on results incl. tables The undiluted NTAN applied with washed and unwashed eyes did not show eye irritation. Overall remarks, attachments Attached background material Attached document Remarks7327-60-8, overview, Dow, 1967, ref.docx / 36.67 KB (application/vnd.openxmlformatsofficedocument.wordprocessingml.document) Applicant's summary and conclusion Interpretation of results not irritating Criteria used for interpretation of results expert judgment Conclusions No irritations was observed following application of the undiluted NTAN.

Endpoint study record: 7372-60-8, Eye irritation, Younger, 1965, SS UUID IUC5-e2574f5d-f79b-4a93-b001-9c9d48658386 Dossier UUID 0 Author LondenK / Akzo Nobel T&E / Arnhem / Netherlands Date 2011-07-14 23:10:06 CEST Remarks

Administrative Data Purpose flag supporting study Study result type experimental result Study period 1965 Reliability 4 (not assignable) Rationale for reliability incl. deficiencies Very limited report only summarizin g the results. The studies were submitted to U.S. EPA under TSCA section 8(e). Data source Reference Referen ce type Author Yea

r Title Bibliograp hic source Testing laborator y Repo rt no. Owner compa ny Compa ny study

no. Repo rt date

study report Young er, F.M. 196 5 Toxicological Investigation Of: NITRILOTRIACETONIT RILE (NTAN) Younger Laboratori

es Y-65- 179 Monsan to 1965- 12-02

Applicant's summary and conclusion Executive summary Onehundred (100.0) milligrams of finely ground sample were placed in the conjunctival sac of the right eye of each of three albino rabbits. observations made over a period of several days for inflamation. The eyes were rinsed with warm isotonic a line solution after twenty-four hours. The compound was classed as a slight eye irritant. The average maximum score as 8.3 out of a possible 110 in one hour.

7.4 Sensitisation Endpoint summary: Sensitisation UUID IUC5-076a3341-27a1-454e-bc38-d69885f047f0 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-11 16:30:57 CET RemarksAdministrative Data Skin sensitisation Short description of key information The OECD toolbox was used to establish skin sensitising properties (QSAR). Key value for chemical safety assessment Skin sensitisation not sensitising Discussion Based on the chemical structure of NTAN it is very unlikely that NTAN has skin sensitising properties. Respiratory sensitisation Short description of key information No information available. Justification for classification or non-classification Based on the QSAR analysis, NTAN does not need classification for skin sensitisation.

7.4.1 Skin sensitisation Endpoint study record: 7327-60-8, Skin sensitisation, Anonymous, 2010, RS UUID IUC5-135e1783-2383-4c6c-ad07-6236af9b11d1 Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 14:31:55 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type (Q)SAR Study period March 2010 Reliability 1 (reliable without restriction) Rationale for reliability incl. deficiencies Generally recogized OECD toolbox was used to establish skin sensitising properties Data source Reference Referenc

e type Author Yea

r Title Bibliographi

c source Testing laborator y Repor t no. Owner compan y Compan y study

no. Repor t date

other company data Anonymou

s 201 0 OECD Toolbox applicatio n study history - CAS # 7327-60-8 Akzo Nobel Functional Chemicals BV Akzo Nobel Functiona l Chemical s BV 2010- 03-26

Data access other: OECD toolbox is publicly available Materials and methods Type of method other: QSAR Type of studyother: OECD toolbox Test guideline Qualifier Guideline Deviations no guideline followed Principles of method if other than guideline QSAR analysis was done using the OECD toolbox. GLP compliance

no Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Test animals Species other: QSAR Strain other: QSAR Sex no data Test system Traditional sensitisation test Route of induction exposure other: QSAR Route of challenge exposure other: QSAR Vehicle other: QSAR Concentration Not applicable No. of animals per dose Not applicable Results and discussion Traditional sensitisation test Results of test (except LLNA) Reading Hours after challenge Group Dose level No. with + reactions Total no. in group Clinical observations

other: QSAR 0 other: QSAR not applicable 0 0 based on chemical structure, skin sensitisation is not very likely Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, NTAN-OECDToolbox-SkinSensitisation, ref.docx / 37.61 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document)Applicant's summary and conclusion Interpretation of results not sensitising Conclusions Based on QSAR, it is very unlikely that NTAN has skin sensitising properties.

7.5 Repeated dose toxicity Endpoint summary: Repeated dose toxicity UUID IUC5-7c599930-7d66-48e8-a38a-61f02b571006 Dossier UUID 0 Author ArtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2011-03-30 11:57:52 CEST Remarks

Administrative Data Short description of key information A combined 28-day oral repeated dose study and reproductive/development study in rats was available. The dose levels for this study were based on a 14-day range-finding study. Key value for chemical safety assessment Repeated dose toxicity: oral

Effect

level NOAEL 10 mg/kg bw/day

Test t y pe subacute

Species rat Discussion 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 th e 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 incidentalchanges commonly observed in rats of this age and strain. Based on the results of this study, the noobserved-adverse-effect level (NOAEL) for systemic toxicity of NTAN was 10 mg/kg/day. the NOAEL in the 14 -day range-finding study was 25 mg/kg bw/day. Justification for classification or non-classification Based on the severe toxicity observed at 45 mg/kg bw (death and severe clinical signs of toxicity), NTAN should be classified with R48/22 and STOT repeated exposure Cat. 2.

7.5.1 Repeated dose toxicity: oral Endpoint study record: 7327-60-8, Repeated dose toxicity: oral, 14-day, Doyle, 2010, SS UUID IUC5-f1f5dd5e-85ea-416a-91a8-bfacfde25380 Dossier UUID 0Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 14:56:19 CET Remarks

Administrative Data

Purpose flag supporting study Study result type experimental result Stud y period July 2009 Reliability 1 (reliable without restriction)

Rationale for reliabilit y incl. deficiencies Well conducted and reported study Data source Reference Referenc

e type Autho

r Yea

r Title Bibliographi

c source Testing laborator y Repor t no. Owner compan y Compan y study

no. Repor t date

study report Doyle A 201 0 14-Day rangefinding oral (gavage) toxicity study of Nitrilotriacetonitril e (NTAN) in rats Experimur , Chicago, IL, USA 09- 570 Akzo Nobel Function al Chemicals LLCChelates 2010- 07-13

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Test type subacute Limit test

no Test guideline Qualifier Guideline Deviations no guideline followed Principles of method if other than guideline range-finding test, primarily according to OECD 407 GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Batch no.: CFC 9161Expiry date: 02 June 2012Purity: 99.6 +/- 1.5%Water: 0.4 +/- 0.2%Test animals Species
rat Strain Sprague-Dawley Sex male/female Administration / exposure Route of administration oral: gavage Vehicle corn oil Analytical verification of doses or concentrations
yes Duration of treatment / exposure 14 days Frequency of treatment daily Doses/concentrations 1, 5, 25, 50 mg/kg bw

Basis actual ingested No. of animals per sex per dose
5 Control animals yes, concurrent vehicle Examinations Any other information on materials and methods incl. tables clinical signs body weight food intake gross necropsy Results and discussions Effect levels Endpoint Effect level Based

on Sex Basis for effect level / Remarks

NOAEL 25 mg/kg bw/day (actual dose received) test mat. male/female clinical signs and death in one male at 50 mg/kg bw Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, repeated dose toxicity, 14-day, Doyle, 2010, ref.docx / 37.83 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Conclusions Based on the results of this study, the no-observed- adverse-effect level (NOAEL) for oral administration of NTAN to Sprague- Dawley rats for 14 consecutive days was 25 mg/kg/day.Executive summary The objective of this study was to evaluate the potent ial toxicity of Nitrilotriacetonitrile (NTAN) following repeated daily oral administration for a period of 14 days and to select dose levels for a longer term study. NTAN was administered by oral gavage as a suspension in a vehicle of corn oil to Sprague-Dawley rats at dose levels of 0 (vehicle only), 1, 5, 25 and 50 mg/kg/day for 14 consecutive days. Each group consisted of 5 rats/sex and were dosed once daily, 7 days/week, for 14 days at a dosing volume of 2 ml/kg. Standard toxicological evaluations included daily clinical observations, weekly body weights and food consumption, and gross evaluations at necropsy. Slight decreases in food consumption, body weight and body weight gain were noted during the first week of treatment in animals receiving NTAN; however, the alterations were transient in nature and generally resolved by the end of the study. Treatment-related clinical signs of toxicity included salivation and red discoloration around the mouth and were noted for one male in the 50 mg/kg group that died spontaneously on day 13 of the study. Treatment-related gross necropsy findings were also noted for this animal and included red fluid in the cranial cavity. All other clinical signs and gross necropsy observations were considered unrelated to treatment with NTAN. Based on the results of this study, the no-observed- adverse-effect level (NOAEL) for oral administration of NTAN to Sprague- Dawley rats for 14 consecutive days was 25 mg/kg/day.

Endpoint study record: 7327-60-8, Repeated dose toxicity: oral, 28-day, Doyle, 2010, RS UUID IUC5-4f3b4c37-b4e3-4ef6-a845-ccf32fe51e0b Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 15:01:19 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period September-November 2009 Reliability 1 (reliable without restriction) Rationale for reliability incl. deficiencies Well conducted and reported study

Data source Reference Referen ce type Auth

or Yea

r Title Bibliograp hic source Testing laborato ry Repo rt no. Owner compan y Compa ny study

no. Repo rt date

study report Doyle A 201 0 Combined oral repeated dose and reproductive/developm ental toxicity screening test of Nitrilotriacetonitrile ( NTAN ) in rats Experim ur, Chicago, IL, USA 09- 574 Akzo Nobel Function al Chemicals LLCChelates 2010- 10-19

Data access data submitter is data owner Data protection claimed yes, but willing to share Cross-reference to same studyA lso used for developmental/reproduction toxicity Materials and methods Test type subacute Limit test

no Test guideline

Qualifier Guideline Deviations according to OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test) Principles of method if other than guideline range-finding test, primarily according to OECD 407 GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Batch no.: CFC 9161Expiry date: 02 June 2012Purity: 99.6 +/- 1.5%Water: 0.4 +/- 0.2% Test animals Species
rat Strain Sprague-Dawley Sex male/female Details on test animals and environmental conditions See further Administration / exposure Route of administration oral: gavage Vehicle corn oil Details on oral exposure Dose formulations of the test material were prepared approximately every 2 weeks as suspensions in a vehicle of corn oil at graded concentrations of 0.5, 5.0 and 22.5 mg/ml, which corresponded to target doses of 1, 10 and 45 mg/kg/day, when administered at a dosing volume of 2 ml/kg. Dose levels of the test material were selected by the Sponsor based on the results of a 14-day range-finding study (Experimur Study No.: 09-570). Test material formulations of NTAN were previously shown to be stable for a minimum of 4 weeks when stored at room temperature. Test material formulations were stored at room temperature (approximately 22 ± 3 degrees C), and thoroughly mixed on a stir plate prior to and during dosing.Analytical verification of doses or concentrations
yes Details on analytical verification of doses or concentrations Samples of the test material formulations were collected from the first preparation (i.e., prior to dosing, week 0), mid-way through the study (approximately week 3), and towards the end of the study (week 4), and analyzed for concentration. Samples for homogeneity (homogeneity is performed on duplicate samples collected from the top, middle and bottom of the test material formulation) were collected from the low and high dose formulations of the 1st preparation and analyzed concurrently with the first concentration analyses. A validated gas chromatography (GC) with flame ionization detection (FID) method was used to determine concentration and homogeneity of the NTAN dose formulations. The analytically-determined concentrations met the acceptance criterion of ± 15% of the target concentration as specified in the study protocol. Samples for homogeneity were collected from the low and high dose formulations at study initiation and were found to be within the protocol-specified acceptance criteria of ± 15%. Duration of treatment / exposure 2 weeks prior to mating, during mating, and until sacrifice (males: 31 doses; females: a minimum of 48 doses) Frequency of treatment daily Doses/concentrations 1, 10, 45 mg/kg bw

Basis actual ingested No. of animals per sex per dose 10 rats/sex main study, 5 additional rats/sex in the control and high dose groups (recovery animals kept for 14 days after the last treatment; these animals were not mated) Control animals yes, concurrent vehicle Details on study design See below Positive control Not used Examinations Observations and examinations performed and frequencyCAGE SIDE OBSERVATIONS: Yes- Time schedule: dailyDETAILED CLINICAL OBSERVATIONS: YesTime schedule: weeklyBODY WEIGHT: Yes - Time schedule for examinations: weeklyFOOD CONSUMPTION: Yes, weekly FOOD EFFICIENCY: NoWATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): NoOPHTHALMOSCOPIC EXAMINATION: NoHAEMATOLOGY: Yes - Time schedule for collection of blood: prior to necropsy- Anaesthetic used for blood collection: sodium pentobarbital- Animals fasted: Yes- How many animals: 5 rats/sex/group (main and recovery)CLINICAL CHEMISTRY: Yes - Time schedule for collection of blood: prior to necropsy- Animals fasted: Yes- How many animals: 5 rats/sex/group (main and recovery)URINALYSIS: NoNEUROBEHAVIOURAL EXAMINATION: Yes - Time schedule for examinations: males after 29 doses, females after post-partum Day 4- Dose groups that were examined:- Battery of functions tested: sensory ac tivity / grip strength / motor activityOTHER: see at developmental/reproduction toxicity Sacrifice and pathology GROSS PATHOLOGY: Yes HISTOPATHOLOGY: Yes Statistics See below Any other information on materials and methods incl. tablesData were manually collected, entered into Excel spreadsheets, and means and standard deviations were calculated using the same software. Data were analyzed for homogeneity of variance using Levene's test. When the variances were homogeneous (p > 0.001), the data were further analyzed by one-way analysis of variance (ANOVA). When a significant F value was observed (p<=0.05) each treatment group was compared to the vehicle control group using Dunnett’s two-tailed t-test. Statistical significance was declared at p<=0.05 for Dunnett’s test. When Levene’s test was significant (p<=0.001), an appropriate transformation was applied to the data(e.g., log-transformation), and the analyses were performed on the transformed data. When the variancestabilizing transformation was not achieved, Kruskal-Wallis test was performed followed by Mann-Whitney U (MWU) statistic. Minimal statistical significance was declared at p<=0.05). Total motor activity data were analyzed using ANOVA, while interval motor activity data were analyzed using repeated measures analysis of variance. All analyses were performed with Systat 10. Minor differences during recalculation of results, as presented in this report, may be noted due to digital storage of numbers in computers or calculators as well as software calculating or rounding issues. Results and discussions Effect levels Endpoint Effect level Based

on Sex Basis for effect level / Remarks

NOAEL 10 mg/kg bw/day (actual dose received) test mat. male/female death, clinical signs, decreased food intake and BW, and motor activity changes at 45 mg/kg bw Results of examinations Clinical signs and mortality
yes Body weight and weight gain
yes Food consumption and compound intake (if feeding study)
yes Food efficiency not examined Water consumption and compound intake (if drinking water study) not examined Ophthalmoscopic examination not examined Haematology no effects Clinical chemistry no effects Urinalysis not examined Neurobehaviour
yes Organ weights no effects Gross pathology
yes Histopathology: non-neoplastic no effectsHistopathology: neoplastic no effects Details on results CLINICAL SIGNS AND MORTALITY: one male and three females were found dead. Clinical signs consisted of discoloration around the mouth and nose, discolored/wet inguinal fur, convulsions, labored breathing, irritability, head weaving, loss of righting reflex, repetitive behavior, cyanosis, salivation, hypoactivity, discolored paws and rough hair coat. Ho wever, these signs were primarily on a single but different day of the study, which is suggestive of the acute nature of this observed toxicity.BODY WEIGHT AND WEIGHT GAIN: reduced at 45 mg/kg bwFOOD CONSUMPTION: reduced at 45 mg/kg bwFOOD EFFICIENCY: not measuredWATER CONSUMPTION: not measured OPHTHALMOSCOPIC EXAMINATION: not measuredHAEMATOLOGY: no treatment-related changesCLINICAL CHEMISTRY: no treatment-related changesURINALYSIS: not measuredNEUROBEHAVIOUR: No treatment-related differences in grip strength or F.O.B. parameters were noted across treatment groups for core or recovery animals. Statistically significant differences in motor activity assessments were observed for males in the 45 mg/kg group, compared to controls, and included decreased totals for move episodes, stereotypy 1 moves and episodes, and stereotypy 2 moves and time, suggestive of a change in pattern of behavior for these animals.The mean values for these parameters generally did not fall within their respective concurrent control ranges and were likely treatment-related. However, this change was only present at the dose level that produced overt systemic toxicity (death). Following the recovery period, total move episodes remained significantly decreased for males of the 45 mg/kg group, compared to controls, indicating that effects on total mo ve episodes were not reversible wi thin the 14-day period following cessation of dosing. No significant differences in motor activity parameters were noted for females across treatment groups for core or recovery animals.ORGAN WEIGHTS: no treatment-related changesGROSS PATHOLOGY: limited to the 45 mg/kg group; 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. Pigmentation of the thymus was also observed in one male of the 45 mg/kg group animal following the recovery period; whereas, one female in the 45 mg/kg group was observed with this finding at the end of the treatment period.HISTOPATHOLOGY: NON-NEOPLASTIC, no treatment-related changes Overall remarks, attachments Overall remarks It should be 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 whic h such changes occur and the poss ibility that damage to other organ systems may contribute to or cause such change s indirectly” (Guidelines for Neurotoxicity Risk Assessment, EPA/630/R-95/001F, April 1998, page 20). 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. Attached background material Attached document Remarks 7327-60-8, repeated dose toxicity, 28-day, Doyle, 2010, ref.docx / 37.81 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Conclusions Based on the results of this study, the no-observed- adverse-effect level (NOAEL) for parental systemic toxicity following oral administration of NTAN to Sprague-Dawley rats for 31 days (males) or at least 48 days (females) was 10 mg/kg/day.Executive summary Nitrilotriacetonitrile (NTAN) was administered to male and female Sprague-Dawley rats by oral gavage at concentrations of 0.5, 5.0 or 22.5 mg/ml equivalent to doses of 1, 10 or 45 mg/kg/day, when administered at a constant dosing volume of 2 ml/kg. The vehicle control group received vehicle (corn oil) only. Ten rats/sex/group were selected for cohabitation (i.e., core/parental animals) and 5 additional rats/sex in the control and high dose groups were designated as recovery animals in order to evaluate reversibility of any NTAN-induced effects. The parental rats were administered NTAN for 2 weeks prior to mating, during the mating period, and until sacrifice (males: 31 doses; females: a minimum of 48 doses). Recovery animals were treated for the same duration as the parental rats, but were not mated, and were held for 14 days after treatment to evaluate recovery from any adverse treatment-related effects. Due to the spontaneous deaths of 2 high dose recovery females, only 3 unmated recovery females remained. In order to increase the size of the female high dose recovery group, one core parental high dose dam was held without treatment for approximately 14 days and was necropsied with the recovery animals. By doing this, there was one additional high dose female (n = 4 instead of n = 3) to evaluate for recovery potential from treatment-related effects. Body weights, body weight gain and food consumption were measured during the study. In addition, clinical observations, a functional observational battery with automated motor activity, organ weights, and clinical and diagnostic pathology were evaluated. Four rats in the 45 mg/kg/day group died spontaneously during the study; male #536 was found dead on study day 19 (during treatment/ cohabitation), female #592 was found dead on day 3 (during treatment/pre-cohabitation), female #599 (designated as a recovery animal) was found dead on day 30 (during the treatment period), and female #586 was found dead on lactation day 3. All four deaths were considered to be treatment-related, although microscopically the cause of death for these animals was undetermined. Treatment-related clinical signs of toxicity were limited to animals in the 45 mg/kg group (6/15 females) and were often noted within a few hours following dose administration. These clinical signs consisted of discoloration around the mouth and nose, discolored/wet inguinal fur, convulsions, labored breathing, irritability, head weaving, loss of righting reflex, repetitive behavior, cyanosis, salivation, hypoactivity, discolored paws and rough hair coat. However, these signs were primarily on a single but different day of the study, which is suggestive of the acute nature of this observed toxicity. Treatment-related decreases in mean food consumption were observed for male and female rats in the 45 mg/kg group during the study. These reductions in food consumption began over study days 0-4 for both sexes and continued for the remainder of the treatment period for females. Corollary reductions in body weight were also observed: mean body weight and/or body weight gain were reduced compared to concurrent control values in both sexes in the 45 mg/kg group beginning over study days 1-4 and these changes generally persisted for males and females throughout the treatment phase of the study. No treatment-related changes in clinical chemistry or hematology profiles were observed in animals treated with NTAN. No treatment-related differences in grip strength or F.O.B. parameters were noted across treatment groups for core or recovery animals, or in motor activity parameters for females. Statistically significant differences in motor activity assessments were observed for males in the 45 mg/kg group, compared to controls, and included decreased totals for move episodes, stereotypy 1 moves and episodes, and stereotypy 2 moves and time, suggestive of a change in pattern of behavior for these animals. Following the recovery period, total move episodes remained significantly decreased for males of the 45 mg/kg group, compared to controls. These changes in motor activity were likely treatment-related but were only present at the dose level that produced overt systemic toxicity (death). Absolute and relative organ weights were not considered to be affected by treatment with NTAN. 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 parental systemic toxicity of NTAN was 10 mg/kg/day.7.6 Genetic toxicity Endpoint summary: Genetic toxicity UUID IUC5-2be2f523-0fe5-4bcb-9e5c-f86d6da8cf38 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-11 16:31:55 CET Remarks

Administrative Data Short description of key information Two in vitro genotoxicity studies were available, an Ames test and a Chromosome Aberration test in CHO cells. Key value for chemical safety assessment Genetic toxicity negative Discussion Both in vitro genotoxicity studies, viz. the Ames test and the chromosome aberration test showed that NTAN was not mutagenic. Justification for classification or non-classification Because both the Ames test and the chromosome aberration test showed that NTAN was not mutagenic, no classification for genotoxicity is needed.

7.6.1 Genetic toxicity in vitro Endpoint study record: 7327-60-8, Genetic toxicity in vitro, Ames, Stankowski 2009, RS UUID IUC5-eedd3cf8-1939-475b-bdc3-ae1d64899cc0 Dossier UUID 0 Author LondenK / Akzo Nobel N.V. / Arnhem / Netherlands Date 2012-01-30 09:56:53 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period May-August 2009 Reliability 1 (reliable without restriction)

Rationale for reliabilit y incl. deficiencies Well conducted and reported study, according to GLP Data source Reference Referenc

e type Author Yea

r Title Bibliographi

c source Testing laborator y Report

no. Owner compan y Compan y study

no. Repor t date

study report Stankows ki LF 200 9 Bacterial

reverse mutation assay with

a confirmator y assay; NTAN Covance Lab. Inc., Vienna, Virgina 22182- 1699, USA 821150 6 Akzo Nobel Functiona l Chemicals LLCChelates, Chicago, 2009- 08-11Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Type of genotoxicity gene mutation Type of study bacterial reverse mutation assay (e.g. Ames test) Test guideline

GLP compliance yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across) yes Test material identity

Details on test material Appearance: off-white crystalline powderPurity: 99 .6 +/- 1.5% Other:- water: 0.4 +/- 0.2% Method Target gene reverse mutations at the histidine locus and tryptophan locus Species/strain Species/strain S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 Metabolic activation with and without Metabolic activation system Aroclor 1254-induced rat liver homogenate Species/strain E. coli WP2 uvr A Metabolic activation with and without Metabolic activation system Aroclor 1254-induced rat liver homogenate Test concentrations In initital test: 1.60, 5.00, 16.0, 50.0, 160, 500, 1600, 5000 μ g/plate with and without S9-mixIn confirmatory test: 50.0, 160, 500, 1600, 5000 μ g/plate with and without S9-mix VehicleDMSO, purity 99.94% Controls

Solvent /

vehicle controls yes (DMSO)

Positive controls yes

Positive control substance other: 2-nitrofluorene, sodium azide, ICR-191, 4-nitroquinoline-N-oxide

Remarks without S9-mix

Solvent /

vehicle controls yes (DMSO)

Positive controls no (benzo(a)pyrene, 2-aminoanthracene)

Remarks with S9-mix Details on test system and conditions Plating Procedures: Each plate was labeled with a code that identified the test article, test phase, tester strain, activation condition and dose level. The S9 mix and dilutions of the test article were prepared immediately prior to their use. Treatments were performed by adding 100 μL tester strain and 50 μL test or controlarticle to 2.5 mL molten selective top agar (maintained at 45 ± 2°C). After the required components had been added, the mixture was vortexed and overlaid onto the surface of 25 mL minimal bottom agar in a 15 x 100 mm petri dish. After the overlay solidified, the plates were inverted and incubated for 52 ± 4 hours at 37 ± 2°C. Cultures were treated in the presence of S9 in an identical manner, except using 2.0 niL undiluted molten selective top agar and adding 500 μL S9 mix.Scoring the Plates: Plates which were not evaluated immediately following the incubation period were held at >0 to 10°C until such time that colony counting and bacterial background lawn evaluation could take place.Bacterial Background Lawn Evaluation: The condition of the bacterial background lawn was evaluated macroscopically and microscopically (using a dissecting microscope) for indications of cytotoxicity and test article precipitate. Evidence of cytotoxicity was scored relative to the vehicle control plate and was recorded along with the revertant counts for all plates at that dose level. Lawns were scored as normal (N), reduced (R), obscured by precipitate (0), macroscopic precipitate present (P), absent (A), or enhanced (E); contaminated plates (C) were also noted.Counting Revertant Colonies: Revertant colonies were counted by automated colony counter and by hand. Evaluation criteria See below at remarks Statistics Not done Any other information on materials and methods incl. tables Assay Acceptance Criteria: Before assay data were evaluated, the criteria for a valid assay had to be met. The following criteria were used to determine a valid assay: Tester Strain Integrity - Histidine/Tryptophan Auxotrophy (his-/trp- Mutation): Salmonella tester strains must exhibit the presence of the his mutation, as demonstrated by the ability to grow on minimal bottom agar plat es supplemented with hi stidine/biotin (but not tryptophan/biotin). Tester strain WP2uvrA must exhibit the presence of the trp mutation, as demonstratedby the ability to grow on bottom agar plates supplemented with tryptophan /biotin (but not histidine/biotin). - rfa Cell Wall Mutation: Salmonella tester strains must exhibit the presence of the rfa mutation, as demonstrated by sensitivity to crystal violet. Tester strain WP2uvrA must exhibit resistance to crystal violet. - pKM101 Plasmid: Tester strains TA98 and TA100 must exhibit the presence of the pKM101 plasmid by demonstrating resistance to ampic illin. In contrast, tester strains T A1535, TA1537, and WP2uvrA must be sensitive to ampicillin. - uvrA and uvrB Mutation: All tester strains must exhibit the presence of the uvrA or uvrB mutations, as demonstrated by their sensitivity to ultraviolet light. Characteristic Number of Spontaneous Revertants The average revertants/plate of the vehicle control cultures must be within the following acceptance limits which are based upon historical data and published reports. TA98: 8 - 60 TAI00: 60 - 240 TA1535: 4 - 45 TA1537: 2 - 25 WP2uvrA: 5 - 40 Tester Strain Culture Density Each culture must reach a target 0D (650(1:4)) = 0.4 to 0.6, which has been demonstrated to be representative of cultures in the late exponential or early stationary phase with ≥ l0E9 cells/mL. Positive Control Values The positive controls must induce a ≥ 3-fold increase in revertants/plate as compared to the concurrent vehicle controls. A 3-fold increase in the absence of S9 indicates that the tester strain can identify a mutagen, while a 3-fold increase in the presence of S9 will be considered to have demonstrated the integrity of the S9 mix as well as the ability of the tester strain to detect a mutagen. Number of Dose Levels A minimum of three non-toxic doses is required to evaluate assay data. Maximum Dose Level The highest dose evaluated was the limit dose of 5000 μg/plate. Assay Evaluation Criteria Once the criteria for a valid assay had been met, responses observed in the assay were evaluated. Criteria for a Positive Response A test article is considered to have produced a positive response if it induces a dose-dependent increase in revertant frequency that is ≥ 2.0-fold vehicle control values for tester strains TA98, TA100, and WP2uvrA, or ≥ 3.0-fold vehicle control values for tester strains TA1535 and TA1537. In addition, any response should be reproducible. Criteriafor a Negative Response A test article is considered to have produced a negative response if no dose-dependent, ≥ 2.0-fold or ≥ 3.0- fold increases are observed in tester strains TA98, TA 100, and WP2uvrA, or TA1535 and TA1537, respectively. Criteriafor an Equivocal Response Even after repeated trials, a test article may produce results that are neither clearly positive nor clearly negative (e.g., responses that do not meet the dose-dependency or fold increase requirements but are reproducible). In those rare instances, the test article may be considered to have produced an equivocal response. Other criteria also may be used in re aching a conclusion about the study results (e.g., comparison to historical control values, biological signifi cance, etc.). In such case s, the Study Director will use sound scientific judgment and clearly report and describe any such considerations. Results and discussions Test results Species/strain S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation with and without

Test s y stem all strains/cell types testedGenotoxicit y negative
C y totoxicit y no, but tested up to limit concentrations

V ehicle controls valid yes

Positive controls valid yes

Species/strain E. coli WP2 uvr A

Metabolic activation with and without

Genotoxicit y negative
C y totoxicit y no, but tested up to limit concentrations

V ehicle controls valid yes

Positive controls valid yes

Additional information on results NTAN was found to form a transparent, non-viscous, colorless solution in DMSO at a concentration of approximately 100 mg/mL, which was the highest doseformulation prepared for treatment; it remained freely soluble at all succeeding lower dilutions prepared assay. Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, Genetic toxicity in vitro, Ames, Stankowski 2009, ref.docx / 38.52 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Interpretation of results negative Conclusions Under the conditions of this test, NTAN did not show mutagenic effects. Executive summary The objective of this study was to evaluate the test article, NTAN (nitrilotriacetonitrile;CAS No.: 7327-60- 8), and/or its metabolites for their ability to induce reverse mutations at the hi stidine locus in several strains of Salmonella (TA98, TAI00, TA1535, and TA1537), and at the tryplophan locus of E. coli strain WP2uvrA, in the presence or absence of an exogenous mammalian metabolic activation system (S9). NTAN was evaluated in the initial mutagenicity assay, in all five tester strains, at doses of 1.60, 5.00, 16.0, 50.0, 160, 500, 1600, and 5000 μg/plate with and without S9. All doses of the test article, as well as the concurrent positive and vehicle controls were evaluated in duplicate plates. Normal growth was observed in all five tester strains, and the test article was found to be freely soluble in the aqueous top agar, at all doses evaluated with and without S9. Revertant frequencies for all doses ofNTAN, in all tester strains with and without S9, approximated those observed in the concurrent vehicle control cultures. NTAN was re-evaluated in the confirmatory mutagenicity assay at doses of 50.0, 160, 500, 1600, and 5000 μg/plate with and without 89. All doses of the test article, as well as the concurrent positive and vehicle controls, were evaluated in triplicate plates. Normal growth again was obse rved in all five tester strains, and the test article again was found to be freely soluble, at all doses evaluated with and without S9. Revertant frequencies for all doses of NTAN, in all tester strains with and without 89, again approximated control values. All positive and vehicle control values were within acceptable ranges, and all criteria for a valid study were met. These results indicate NTAN was negative in the Bacterial Reverse Mutation Assay with a ConfirmatoryAssay under the conditions, and according to the criteria, of the test protocol.

Endpoint study record: 7327-60-8, Genetic toxicity in vitro, CAT, Murli, 2009, RS UUID IUC5-10407bb7-94ed-476d-b431-393e13bcf509 Dossier UUID 0 Author GeltenP / Akzo Nobel T&E / Arnhem / Netherlands Date 2010-12-21 15:10:50 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period May- A ugust 2009 Reliability 1 (reliable without restriction)

Rationale for reliabilit y incl. deficiencies Well conducted and reported study according to GLP Data source Reference Referenc

e type Autho

r Yea

r Title Bibliographi

c source Testing laboratory Report

no. Owner compan y Compan y study

no. Repor t date

study report Murli H 200 9 Chromosom al aberrations in Chinese hamster ovary (CHO) cells - NTAN Covance Laboratorie s Inc., Vienna, Virginia, USA 821150 5 Akzo Nobel Functiona l Chemicals LLCChelates 2009- 11-02

Data access data submitter is data owner Data protection claimed yes, but willing to share Materials and methods Type of genotoxicity chromosome aberration Type of study in vitro mammalian chromosome aberration test Test guideline

Qualifier Guideline Deviations according to OECD Guideline 473 (In vitro Ma mmalian Chromosome Abe rration Test) GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier IdentityCAS number 7327-60-8 Details on test material Batch no.: CFC 9161Expiry date: 2 June 2010Purity: 99.6 +/- 1.5%Water: 0.4 +/- 0.2% Method Target gene Chromosome aberration Species/strain Species/strain Chinese hamster Ovary (CHO)

Details on mammalian cell lines ( if applicable ) permament cell line (University of California, San Francisco, USA)

Additional strain characteristics not applicable

Metabolic activation with and without

Metabolic activation

s y stem Aroclor 1254-induced rat liver (S9-mix)

Test concentrations Initital test: - concentrations tested: 18-2700 μg/mL- concentrations evaluated: 1320, 1890, 2700 μg/mL (- S9 mix)- concentrations evaluated: 926, 1320, 1890 μg/mL (+ S9 mix)Confirmatory test: - concentrations tested: 61-2620 μg/mL- concentrations evaluated: 1940, 2280, 2620 μg/mL (-S9 mix)- concentrations evaluated: 1940, 2280, 2620 μg/mL (+ S9 mix) Vehicle DMSO Controls

Ne g ative controls no

Solvent /

vehicle controls yes

True

ne g ative controls no

Positive controls yes

Positive control substance other: mitomycin (-S9 mix) cyclophosphamide (+S9 mix)

Details on test system and conditions Initial test: 3 h exposure, harvest at ~20 h (both with and without S9 mix)Confirmatory test: 20 h exposure, harvest at ~20 h (without S9 mix); 3 h exposure, harvest at ~20 h (with S9 mix) Evaluation criteria See below StatisticsSee below Any other information on materials and methods incl. tables Data Evaluation Mitotic index was evaluated from the vehicle controls and a range of concentrations by analyzing the number of mitotic cells in 1000 cells, if possible, and the ratio expressed as a percentage of mitotic cells. When analysis for aberrations was performed, 100 cells, if possible, from each replicate culture from three concentrations of the test article, vehicle controls, and one dose of the positive control cultures were analyzed for the different types of chromosomal aberrations. At least 25 cells were analyzed from those cultures that had greater than 25% of cells with one or more aberrations. Cells were selected for good morphology and only cells with the number of centromeres equal to the modal number 21 ± 2 (range 19- 23) were analyzed. Percent polyploidy and endoreduplication were also analyzed from the dose levels selected for analysis and controls by evaluating at least 100 metaphases, if possible, and tabulated. For control of bias, all slides were coded prior to analysis. Cells with aberrations were recorded on the data sheets by the microscope stage location. Data Presentation Data were summarized in tables showing mitotic index and percent cells with polyploidy and endoreduplication. For structural aberrations, the aberrant cells were presented in two categories: the number and percentages of aberrant cells excluding gaps (-g), and the number and percentages of aberrant cells including gaps (+g). Assay Acceptance Criteria An assay was considered acceptable for evaluation of test results only if all of the following criteria were satisfied. The metabolic activation and nonactivation sections of the aberrations assay were independent units and would be repeated independently, as needed, to satisfy the acceptance criteria. Acceptable Controls The vehicle control cultures must contain less than —5% cells with aberrations. The positive control result must be significantly higher (p<=0.01) than the vehicle controls. Acceptable High Dose If the aberration results are negative and there is no significant reduction (~>=50%) in mitotic index, the assay must include the highest applicable dose (a target dose of 10mM or 5 mg/mL, whichever is lower) or a dose exceeding the solub ility limit in culture medium. Acceptable Number of Doses The assay must include at least three analyzable concentrations. Assay Evaluation Criteria After completion of microscopic analysis for aberrations, data were decoded. The following factors are taken into account in evaluation of the test article data: • The number and percentages of aberrant cells excluding gaps (-g). • The number and percentages of aberrant cells including gaps (+g). • Evidence of a dose-response relationship. The experimental unit is the cell, and therefore the percentage of cells with structural aberrations was the basis for evaluation. Statistical analysis employed a Cochran Armitage test for linear trend and Fisher’s Exact Test to compare the percentage of cells with a berrations in treated cells to the results obtained for the vehicle controls. Statistical analysis was also performed for cells exhibiting polyploidy and/or endoreduplication in order to detect significant (p ≤ 0.01) increases in these events as indicators of possible induction of numerical aberrations. Evaluation of a Positive Response A test article was considered positive for inducing chromosomal aberrations if a significant increase (the difference was considered significant when p ≤ 0.01) in the number of cells with chromosomal aberrations is observed at one or more concentrations. The linear trend test evaluated the dose-responsiveness. A dose-response should be observed if a significant increase was seen at one or more concentrations. Evaluation of a Negative Response A test article was considered negative for inducing chromosomal aberrations if no significant increase was observed in the number of cells with chromosomal aberrations at any of the concentrations. Equivocal Evaluation Although most assays give clearly positive or negative results, in rare cases the data set would preclude making a defmitive judgment about the activity of the test article. Results might remain equivocal orquestionable regardless of the number of times the assay is repeated. Although the evaluation criteria provided here is normally sufficient, the Study Director might use additional considerations to obtain a fmal evaluation of the test article based upon the Study Director’s scientific judgment. Results and discussions Test results Species/strain Chinese hamster Ovary (CHO)

Metabolic activation without

Genotoxicit y negative
C y totoxicit y yes at 2620 μg/mL (20 h treatment)

V ehicle controls valid yes

Ne g ative controls valid not examined

Positive controls valid yes

Species/strain Chinese hamster Ovary (CHO)

Metabolic activation with

Genotoxicit y negative
C y totoxicit y yes (at >= 2280 μg/mL in confirmatory test)

V ehicle controls valid yes

Negative controls valid not examined

Positive controls valid yes

Overall remarks, attachments Attached background material Attached document Remarks 7327-60-8, genetic toxicity in vitro, CAT, Murli, 2009, ref.docx / 38 KB (application/vnd.openxmlformats-officedocument.wordprocessingml.document) Applicant's summary and conclusion Interpretation of results negative Conclusions NTAN was considered negative for inducing chromo somal aberrations in CHO cells with and without metabolic activation. Executive summary The objective of this in vitro assa y was to evaluate the ability of NTAN (Nitrilotriacetonitrile; CAS No.: 7327-60-8) to induce chromosomal aberrations in cultured Chinese hamster ovary (CHO) cells without and with an exogenous metabolic activation system. Dimethylsulfoxide (DMSO) was the vehicle for the study. The highest concentration tested in the assay was 2700 μgImL, which was above the solubility limit of the formulated NTAN after dosing into culture medium. The stock formulations and their dilutions were dosed using a dosing volume of 1% (10.0 μL/mL) and the vehicle control cultures were treated with 10.0 μL/mL of DMSO. In the initial chromosomalaberrations assay, the treatment period was for 3 hours without and with metabolic activation and the cultures were harvested ~20 hours from the initiation of treatment. Concentrations of 18.3, 26.2, 37.4, 53.4, 76.3, 109,156, 222, 318, 454, 648, 926, 1320, 1890, and 2700 μg/mL were tested without and with metabolic activation. Cultures treated with concentrations of 1320, 1890, and 2700 μtg/mL without metabolic activation and 926, 1320, and 1890 μtg/mL with metabolic activation were analyzed for chromosomal aberrations. No significant increase in cells with chromosomal aberrations, polyploidy, or endoreduplication was observed in the cultures analyzed. In the confirmatory chromosomal aberrations assay, the treatment period was for ~ 20 hours without metabolic activation and 3 hours with metabolic activation and the cultures were harvested ~ 20 hours from the initiation of treatment. Concentrations of 60.7, 121, 243, 485, 728, 971, 1210, 1460, 1940, 2280, and 2620 μg/mL were tested without metabolic activation and 485, 971, 1210, 1460, 1940, 2280, and 2620 μg/mL were tested with metabolic activation. Cultures treated with concentrations of 1940, 2280, and 2620 μg/mL without and with metabolic activation were analyzed for chromosomal aberrations. No significant increase in cells with chromosomal aberrations, polyploidy, or endoreduplication was observed in the cultures analyzed. The vehicle control cultures were in the historical control range for cells with chromosomal aberrations and the positive control cultures had significant increase in cells with chromosomal aberrations as compared with the vehicle control cultures. The high doses selected for analysis in the assay had >50% reduction in mitotic index as compared to the vehicle control cultures and/or a precipitate at the end of the treatment period as recommended for this assay by the OECD Testing Guidelines. The test article, NTAN, was considered negative for inducing chromosomal aberrations in cultured CHO cells without and with metabolic activation.

7.8 Toxicity to reproduction Endpoint summary: Toxicity to reproduction, UUID IUC5-6c49f5a5-b56d-4c65-8ec0-f6054f181b96 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-11 16:32:34 CET Remarks

Administrative Data Effects on fertility Short description of key information A combined oral repeated dose (subacute) and reproductive/developmental toxicity screening study was available. Discussion 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 (see also the 28-day repeated dose study). Overall mating, pregnancy, offspring survival and pup body weights were not notably affected by treatment with NTAN. Based on the results of this study, the no-observed-adverse-effect level (NOAEL) for parental systemic toxicity of NTAN was 10 mg/kg/day and the NOAEL for reproductive/developmental toxicity was at least 45 mg/kg/day. Developmental toxicity / teratogeniticy Short description of key information In the combined oral repeated dose (subacute) and reproductive/developmental toxicity screening study, it was noted (see above) that offspring survical and pup body weights were not notably affected by treatment with NTAN. Gross external observations, fo llowing 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 externalobservations were attributed to NTAN treatment.Visceral examinations were performed on dead pups, when feasible. No visible abnormalities were noted during the visceral examinations. Discussion Although not a full-blown developmental toxicity study was available, the information described above indicates that NTAN did not induce 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. Toxicity to reproduction: other studies Short description of key information No info Justification for classification or non-classification NTAN did not show reproduction/developmental toxicity at a level up to 45 mg/kg bw, although this level resulted in severe parental toxicity, including death. As such no classification for reproduction toxicity is warranted.

7.8.1 Toxicity to reproduction Endpoint study record: 7327-60-8, Toxicity to reproduction, Doyle, 2010, RS UUID IUC5-7ea14391-1a96-492e-b395-77b478fb53d2 Dossier UUID 0 Author A rtsJ / Akzo Nobel Functional Chemicals BV / Amersfoort / Netherlands Date 2010-11-10 12:10:27 CET Remarks

Administrative Data

Purpose flag key study; robust study summary Study result type experimental result Study period September-November 2009 Reliability 1 (reliable without restriction) Rationale for reliability incl. deficiencies Well conducted and reported study under GLP

Data source Reference Referen ce type Auth

or Yea

r Title Bibliograp hic source Testing laborato ry Repo rt no. Owner compan y Compa ny study

no. Repo rt date

study report Doyle A 201 0 Combined oral repeated dose and reproductive/developm ental toxicity screening test of Nitrilotriacetonitrile (NTAN) in rats Experim ur, Chicago, IL, USA 09- 574 Akzo Nobel Function al Chemicals LLCChelates 2010- 10-19

Data access data submitter is data owner Data protection claimed yes, but willing to share Cross-reference to same studyStusy also used for repeated exposure and developmental toxicity Materials and methods Test type screening Limit test

no Test guideline

Qualifier Guideline Deviations according to OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test) GLP compliance
yes Test materials Identity of test material same as for substance defined in section 1 (if not read-across)
yes Test material identity

Identifier Identity

CAS number 7327-60-8 Details on test material Batch no.: CFC 9161Expiry date: 02 June 2012Purity: 99.6 +/- 1.5%Water: 0.4 +/- 0.2% Test animals Species
rat Strain Sprague-Dawley Sex male/female Details on test animals and environmental conditions See further Administration / exposure Route of administration oral: gavage Vehicle corn oil Details on exposure Dose formulations of the test material were prepared approximately every 2 weeks as suspensions in a vehicle of corn oil at graded concentrations of 0.5, 5.0 and 22.5 mg/ml, which corresponded to target doses of 1, 10 and 45 mg/kg/day, when administered at a dosing volume of 2 ml/kg. Dose levels of the test material were selected by the Sponsor based on the results of a 14-day range-finding study (Experimur Study No.: 09-570). Test material formulations of NTAN were previously shown to be stable for a minimum of 4 weeks when stored at room temperature. Test material formulations were stored at room temperature (approximately 22 ± 3 degrees C), and thoroughly mixed on a stir plate prior to and during dosing. Details on mating procedure One male was housed with one female of the same dose level for up to 2 weeks. Daily vaginal smears, taken from the female rats during the mating period, were evaluated for the presence of sperm as well as the stage of the estrus cycle. The presence of sperm in the smear or a sperm-plug indicated a positivemating, and that day was designated as gestation day 0. Ten (10) spermpositive female rats per group were selected to produce the F1 generation. Analytical verification of doses or concentrations
yes Details on analytical verification of doses or concentrations Samples of the test material formulations were collected from the first preparation (i.e., prior to dosing, week 0), mid-way through the study (approximately week 3), and towards the end of the study (week 4), and analyzed for concentration. Samples for homogeneity (homogeneity is performed on duplicate samples collected from the top, middle and bottom of the test material formulation) were collected from the low and high dose formulations of the 1st preparation and analyzed concurrently with the first concentration analyses. A validated gas chromatography (GC) with flame ionization detection (FID) method was used to determine concentration and homogeneity of the NTAN dose formulations. The analytically-determined concentrations met the acceptance criterion of ± 15% of the target concentration as specified in the study protocol. Samples for homogeneity were collected from the low and high dose formulations at study initiation and were found to be within the protocol-specified acceptance criteria of ± 15%. Duration of treatment / exposure 2 weeks prior to mating, during mating, and until sacrifice (males: 31 doses; females: a minimum of 48 doses) Frequency of treatment Daily Details on study schedule See above Doses / concentrations 1, 10, 45 mg/kg bw/day

Basis actual ingested No. of animals per sex per dose 10 rats/sex main study, 5 additional rats/sex in the control and high dose groups (recovery animals kept for 14 days after the last treatment; these animals were not mated) Control animals yes, concurrent vehicle Positive control Not used Examinations Parental animals: Observations and examinations CAGE SIDE OBSERVATIONS: Yes DETAILED CLINICAL OBSERVATIONS: Yes BODY WEIGHT: Yes FOOD CONSUMPTION: YesWATER CONSUMPTION: Nosee for furthet details and outcome, the repeated dose toxicity study (28-day). Estrous cyclicity (Parental animals) Daily vaginal smears were collected over 3 days prio r to cohabitation to ensure and evaluate cyclicity. Vaginal smears were collected during the mating period until the presence of sperm was detected. Sperm parameters (Parental animals) No Litter observations Physical Examination of Rat Pups: Rat pups (F1 generation) were sexed and given a gross external physical examination on postnatal day 0 and again on postnatal day 4. Dead or stillborn pups were examined for gross external anomalies, and, when feasible (for fresh un-autolyzed pups), a visceral examination was performed.Postmortem examinations (Parental animals) All animals, including those found dead, received a complete necropsy. Necropsy included examination of the external surface of the body, all orifices, the cranial, thoracic, and peritoneal cavities, and their contents. The uterus of each rat was examined for evidence of implantation sites and the number of sites was recorded; in addition, the corpora lutea were counted. Rats scheduled for necropsy were fasted overnight and were euthanized using sodium pentobarbital. Seevral tissues were collected. In addition, a bone marrow smear was collected and fixed for possible future evaluation. Testes and epididymides were fixed in Bouin’s solution, and the eyes and associated tissues (e.g., optic nerve and Harderian gland) were fixed in Davidson’s solution; all other tiss ues were fixed in10% neut ral buffered formalin. Postmortem examinations (Offspring) Only in case of dead or stillborn pups (see above). Statistics See below Reproductive indices See at results Offspring viability indices See at results Any other information on materials and methods incl. tables Data were manually collected, entered into Excel spreadsheets, and means and standard deviations were calculated using the same software. Data were analyzed for homogeneity of variance using Levene's test. When the variances were homogeneous (p > 0.001), the data were further analyzed by one-way analysis of variance (ANOVA). When a significant F value was observed (p<=0.05) each treatment group was compared to the vehicle control group using Dunnett’s two-tailed t-test. Statistical significance was declared at p<=0.05 for Dunnett’s test. When Levene’s test was significant (p<=0.001), an appropriate transformation was applied to the data(e.g., log-transformation), and the analyses were performed on the transformed data. When the variancestabilizing transformation was not achieved, Kruskal-Wallis test was performed followedby Mann-Whitney U (MWU) statistic. Minimal statistical significance was declared at p<=0.05). ChiSquare analysis was not warranted for the reproductive performance parameters as mating incidence was relatively identical across groups. Minor differences during recalculation of results, as presented in this report, may be noted due to digital storage of numbers in computers or calculators as well as software calculating or rounding issues. Results and discussions Effect levels Endpoint Generation Sex Effect level Based

on Basis for effect level / Remarks NOAEL P male/female >= 45 mg/kg bw/day (actual dose received) test mat. No effects on reproduction observed Results of examinations: parental animals Clinical signs (parental animals)
yes Body weight and food consumption (parental animals)
yes Test substance intake (parental animals) no effects Reproductive function: estrous cycle (parental animals) not examined Reproductive function: sperm measures (parental animals) not examinedReproductive performance (parental animals) no effects Organ weights (parental animals) no effects Gross pathology (parental animals)
yes Histopathology (parental animals) no effects Details on results (parental animals) See for general toxicity, the repeated dose study (28-day). No differences were detected in the percentage of breeding pairs that mated across the groups (100% in all groups). All females were detected as sperm positive during the mating period, with the exception of animal #575 in the 1 mg/kg group for which no evidence of mating was detected due tomisdiagnosis of a vaginal smear; this animal was pregnant, underwent successful parturition and was therefore considered to be sperm positive. Thenumber of dams that successfully underwent parturition was 9-10 in each group. One female (#558) in the vehicle control group was detected as sperm positive, but did not show any evidence of parturition or pregnancy. Another female (#581) in the 10 mg/kg group was pregnant (as detected byimplantation sites in the uterus), but never showed signs/evidence of parturition. The average number of days to achieve sperm positive status ranged from 3.5-4.1 days across treatment groups. The length of gestation was similar across all groups and averaged approximately 22 days. No significant differences in the mean number of corpora lutea/dam or total implantation sites/dam were observed. Results of examinations: offspring Viability (offspring) no effects Clinical signs (offspring) no effects Body weight (offspring) no effects Sexual maturation (offspring) not examined Organ weights (offspring) not examined Gross pathology (offspring) not examined Histopathology (offspring) not examined Details on results (offspring) Evaluations of litter survival and offspring body weights were limited to those dams that underwent successful parturition. No statistically significant di fferences in mean litter size were detected across groups. The number of dead fetuses that were observed (ranged from 2-5/group) and the percentage of pups surviving to lactation day 4 were comparable across groups. In addition, no significant differences in pre-implantation, pre-natal or post-natal loss of offspring were observed across groups. Mean sex ratios (males/females) for days 0 and 4 were also unaffected by treatment. No significant differences in offspring body weights were observed. 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 (of the F1 generation) in each treatment group, including the vehicle control group. One control fetus was noted with a blunted tail on lactation days 0 and 4. All other gross external morphology observations were common observations likely associated with delivery and consisted mainly of bruising, pale skin and a lesion/scabon the neck. Two pups in the 1 mg/kg group were noted as cannibalized on day 0; one of which was missing its left hindlimb and tail (most likely due to cannibalization). 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 stomach of one pup from dam# 590 in the 45 mg/kg group was noted to be devoid of milk; however, this observation was not attributed to NTAN treatment. Applicant's summary and conclusion Conclusions 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 (see also the 28-day repeated dose study). Overall mating, pregnancy, offspring survival and pup body weights were not notably affected by treatment with NTAN. Based on the results of this study, the no-observed-adverse-effect level (NOAEL) for parental systemic toxicity of NTAN was 10 mg/kg/day and the NOAEL for reproductive/developmental toxicity was at least 45 mg/kg/day. Executive summary Nitrilotriacetonitrile (NTAN) was administered to male and female Sprague-Dawley rats by oral gavage at concentrations of 0.5, 5.0 or 22.5 mg/ml equivalent to doses of 1, 10 or 45 mg/kg/day, when administered at a constant dosing volume of 2 ml/kg. The vehicle control group received vehicle (corn oil) only. Ten rats/sex/group were selected for cohabitation (i.e., core/parental animals) and 5 additional rats/sex in the control and high dose groups were designated as recovery animals in order to evaluate reversibility of any NTAN-induced effects. The parental rats were administered NTAN for 2 weeks prior to mating, during the mating period, and until sacrifice (males: 31 doses; females: a minimum of 48 doses). Recovery animals were treated for the same duration as the parental rats, but were not mated, and were held for 14 days after treatment to evaluate recovery from any adverse treatment-related effects. Due to the spontaneous deaths of 2 high dose recovery females, only 3 unmated recovery females remained. In order to increase the size of the female high dose recovery group, one core parental high dose dam was held without treatment for approximately 14 days and was necropsied with the recovery animals. By doing this, there was oneadditional high dose female (n = 4 instead of n = 3) to evaluate for recovery potential from treatmentrelated effects. Body weights, body weight gain and food consumption were measured during the study. In addition, clinical observations, a functional observational battery with automated motor activity, organ weights, and clinical and diagnostic pathology were evaluated. Four rats in the 45 mg/kg/day group died spontaneously during the study; male #536 was found dead on study day 19 (during treatment/ cohabitation), female #592 was found dead on day 3 (during treatment/pre-cohabitation), female #599 (designated as a recovery animal) was found dead on day 30 (during the treatment period), and female #586 was found dead on lactation day 3. All four deaths were considered to be treatment-related, although microscopically the cause of death for these animals was undetermined. Treatment-related clinical signs of toxicity were limited to animals in the 45 mg/kg group (6/15 females) and were often noted within a few hours following dose administration (see also the 28 -day repeated toxicity study). Treatment-related decreases in mean food consumption were observed for male and female rats in the 45 mg/kg group during the study. These reductions in food consumption began over study days 0-4 for both sexes and continued for the remainder of the treatment period for females. Corollary reductions in body weight were also observed: mean body weight and/or body weight gain were reduced compared to concurrent control values in both sexes in the 45 mg/kg group beginning over study days 1-4 and these changes generally persisted for males and females throughout the treatment phase of the study. No treatment-related changes in clinical chemistry or hematology profiles were observed in animals treated with NTAN (see also the 28 -day repeated toxicity study). No differences were detected in the percentage of breeding pairs that mated across the groups (100% in all groups). The number of dams that successfully underwent parturition was 9-10 in each group. No gross external or visceral observations were attributed to NTAN treatment. Overall mating, pregnancy, offspring survival and pup body weights were not notably affected by treatment with NTAN.Absolute and relative organ weights were not considered to be affected by treatment with NTAN. 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 parental systemic toxicity of NTAN was 10 mg/kg/day and the NOAEL for reproductive/developmental toxicity was 45 mg/kg/day.