1 / 89



# I U C L I D

Data Set

Existing Chemical : ID: 124

- 68

- 5

CAS No. : 124

- 68

- 5

EINECS Name : 2

- amino

- 2

- methylpropanol

EC No. : 204

- 709

- 8

TSCA Name : 1

- Propanol, 2

- amino

- 2

- methyl

Molecular Formula : C4H11NO

Producer related part Company : Dow Chem ical, TERC

Creation date : 01.03.2004

Substance related part Company : Dow Chemical, TERC

Creation date : 01.03.2004

Status : Memo : AMP

Printing date : 30.11.2006

Revision date : Date of last update : 30.11.2006

Number of pages : 89

Chapter (profile) : Chapter: 1, 2, 3, 4, 5, 6, 7, 8, 10 Reliability (profile) : Reliability: without reliability, 1, 2, 3, 4 Flags (profile) : Flags: without flag, confidential, non confidential, WGK (DE), TA

- Luft (DE), Material Safety Dataset, Ri sk Assessment, Directive 67/548/EEC, SIDS

## 1. General Information Id 124

- 68

- 5

Date 30.11.2006

2 / 89 1.0.1 APPLICANT AND COMPANY INFORMATION

Type : manufacturer

Name : The Dow Chemical Company Contact person : Dr. William T. Stott

Date : 27.04.2005

Street : 1803 Building

Town : 48674 Midland, MI

Coun try : United States

Phone : 989 636 1000

Telefax : Telex : Cedex : Email : Homepage : http://www.dow.com/Homepage/index.html

27.04.2005

1.0.2 LOCATION OF PRODUCTION SITE, IMPORTER OR FORMULATOR

Type : manufacturer Name of plant : ANGUS Chemical Company

Street : Town : Sterlington, Louisiana Country : United States

Phone : Telefax : Telex : Cedex : Email : Homepage : http://www.dow.com/Homepage/index.html

Source : Frauson, L. (2003). Internal E

- mail of the Dow Chemical Company to C.

Houtman. Reliability : (2) valid with restrictions 2g

27.04.2005

Type : manufacturer Name of plant : Angus Chemie GmbH

Street : Town : Ibbenburen Country : Germany

Phone : Telefax : Telex : Cedex : Email : Homepage : http://www.dow.com/Homepage/index.html

Source : Frauson, L. (2003). Internal E

- mail of the Dow Chemical Company to C.

Houtman. Reliability : (2) valid with restrictions 2g

27.04.2005

## 1. General Information Id 124

- 68

- 5

Date 30.11.2006

3 / 89 1.0.3 IDENTITY OF RECIPIENTS

1.0.4 DETAILS ON CATEGORY/TEM PLATE

1.1.0 SUBSTANCE IDENTIFICA TION

IUPAC Name : 2

- Amino

- 2

- methyl

- 1

- propanol

Smiles Code : OCC(N)(C)C

Molecular formula : C4H11NO Molecular weight : 89.14

Petrol class :

Reliability : (2) valid with restrictions 2g

28.06.2004 (1)

1.1.1 G ENERAL SUBSTANCE INFORMATION

Purity type : measured for specific batch Substance type : organic Physical status : liquid Purity : ca. 100 % w/w

Colour : colourless

Odour : faint amine odor

Remark : The test material is sold in formulations of >99% ai, 95% ai, 90% ai, and

75% ai.

AMP

- Regular is 92% minimum ai, maximum 8% impurities.

Source : MSDS of The Dow Chemical Company. 2003. Reliability : (2) valid with restrictions 2g

27.04.2005

1.1.2 SP ECTRA

1.2 SYNONYMS AND TRADENA MES

2

- Amino

- 2

- methyl

- 1

- propanol

Reliability : (2) valid with restrictions

18.03.2004 (1)

Aminomethylpropanol

Source : ANGUS Chemie GmbH Ibbenburen European Commission

- European Chemicals Bureau Ispra(VA)

Relia bility : (2) valid with restrictions

01.03.2004

AMP

Source : ANGUS Chemie GmbH Ibbenburen

## 1. General Information Id 124

- 68

- 5

Date 30.11.2006

4 / 89 European Commission

- European Chemicals Bureau Ispra(VA) Environmental Evaluation Report, DR

- 0001

- 9476. The Dow Chemical Company. Reliability : (2) valid w ith restrictions

01.03.2004

AMP 100R

27.04.2005

AMP

- 75

27.04.2005

AMP

- 90

27.04.2005

AMP

- 90 Textile

27.04.2005

AMP

- 95

Source : Environmental Evaluation Report, DR

- 0001

- 9476. The Dow Chemical Company. Reliability : (2) valid with restrictions

01.03.2004

AMP

- 95 Diagnostic

27.04.2005

AMP

- 95 SG

27.04.2005

AMP

- Regular Diagnostic

27.04.2005

AMP

- ULTRA PC 1000 Neutralizing Amine

27.04.2005

AMP

- ULTRA PC 2000 Neutralizing Amine

27.04.2 005

AVT

- 75 Amino Alcohol

27.04.2005

Corrguard

- 100 Animo Alcohol

27.04.2005

Corrguard

- 75 Animo Alcohol

27.04.2005

Corrguard

- 95 Animo Alcohol

## 1. General Information Id 124

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- 5

Date 30.11.2006

5 / 89 27.04.2005

Corrguard

- A Animo Alcohol

27.04.2005

P

- 1826

27.04.2005

1.3 IMPURITIES

Purity : typical for marketed substance

CAS

- No : 124

- 68

- 5

EC

- No : 204

- 709

- 8

EINECS

- Name : 2

- amino

- 2

- methylpropanol

Molecular formula : C4H11NO

Value : 75

- 100 % w/w

Source : MSDS o f The Dow Chemical Company. 2003. Reliability : (2) valid with restrictions 2g

18.11.2004

Purity : typical for marketed substance

CAS

- No : 27646

- 80

- 6

EC

- No : EINECS

- Name : MMAMP

Molecular formula : Value : < 6.9 % w/w

27.04.2005

1.4 ADDITIVES

Purity type : other

CAS

- No : 7732

- 18

- 5

EC

- No : 231

- 791

- 2

EINECS

- Name : water

Molecular formula : H2O

Value : 0

- 25 % w/w

Function of additive : S olvent

Source : MSDS of The Dow Chemical Company. 2003. Reliability : (2) valid with restrictions 2g

01.03.2004

1.5 TOTAL QUANTITY

1.6.1 LABELLING

Labelling : as in Directive 67/548/EEC Specific limits : yes

## 1. General Information Id 124

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- 5

Date 30.11.2006

6 / 89 Symbols : Xi, , ,

Nota : C,

, R

- Phrases : (36/38) Irritating to eyes and skin (52) Harmful to aquatic organisms (53) May cause long

- term adverse effects in the aquatic environment

S

- Phrases : (2) Keep out of reach of children (61) Avoid release to the environment. Refer to specia l instructions/Safety

data sets

Source : European Commission

- European Chemicals Bureau Ispra (VA)

01.03.2004

1.6.2 CLASSIFICATION

Classified : as in Directive 67/548/EEC Class of danger : irritating

R

- Phrases : (36/38) Irritating to eyes and skin (52) Harmful to aquatic organisms (53) May cause long

- term adverse effects in the aquatic environment Specific limits :

Remark : Phrases apply for concentrations of AMP > 25%.

Source : European Commission

- European Chemicals Bureau Ispra (VA)

27.04.2005

1.6.3 PACKAGING

1.7 USE PATTERN

Type of use : industrial Category : Paints, lacquers and varnishes industry

27.04.2005

Type of use : industrial Category : other: paper coating, personal care, metalworking fluid

Reliability : (2) valid with restrictions 2g

01.03.2004 (1)

1.7.1 DETAILED USE PATTERN

1.7.2 METHODS OF MANUFACTURE

1.8 REGULATORY MEASURES

1.8.1 OCCUPATIONAL EXPOSURE LIMIT VALUES

Type of limit : other

## 1. General Information Id 124

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Date 30.11.2006

7 / 89 Limit value :

Remark : none established

Source : ANGUS Chemie GmbH Ibbenburen European Commission

- European Chemicals Bureau Ispra (VA)

01.03.2004

1.8.2 ACCEPTABLE RESIDUES LEVELS

1.8.3 WATER POLLUTION

1.8.4 MAJOR ACCIDENT HAZARDS

1.8.5 AIR POLLUTION

1.8.6 LISTINGS E.G. CHEMICAL INVENTORIES

1.9.1 DEGRADATION/TRANSFORMATION PRODUCTS

1.9.2 COMPONENTS

1.10 SOURCE OF EXPOSURE

Source of exposure : Human: exposure by production Exposure to the : Substance

Source : Frauson, L. (2003). Internal E

- mail of the Dow Chemical Company to C.

Houtm an. Reliability : (2) valid with restrictions 2g

01.03.2004

Source of exposure : Human: exposure of the consumer/bystander Exposure to the : Substance

Source : Frauson, L. (2003). Internal E

- mail of the Dow Chemical Company to C.

Houtman.

R eliability : (2) valid with restrictions 2g

01.03.2004

1.11 ADDITIONAL REMARKS

1.12 LAST LITERATURE SEARCH

## 1. General Information Id 124

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Date 30.11.2006

8 / 89 1.13 REVIEWS

## 2. Physico

- Chemical Data Id 124

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- 5

Date 30.11.20 06

9 / 89 2.1 MELTING POINT

Value : 30.5 °C Decomposition : no, at 31 °C

Sublimation : no Method : other: unknown

Year : 1989

GLP : no data

Test substance : as prescribed by 1.1

- 1.4

Reliability : (2) valid with restrictions 2 (data from a collection) Flag : Critical study for SID S endpoint

10.05.2006 (2)

2.2 BOILING POINT

Value : 165.5 °C at 1013 hPa Decomposition : no Method : other

Year : 1989

GLP : no data

Test substance : as prescribed by 1.1

- 1.4

Reliability : (2 ) valid with restrictions 2 (data from a collection) Flag : Critical study for SIDS endpoint

10.05.2006 (2)

2.3 DENSITY

Type : relative density

Value : = .934 at 40 °C

Method : other:unknown

Ye ar : 1989

GLP : no Test substance : as prescribed by 1.1

- 1.4

Source : Budavari, S. (1989) The Merck Index (Eleventh Edition). Merck & Co., INC. Rahway, NJ, USA. Not GLP, Published Reliability : (2) valid with restrictions 2 (data from a collect ion) Flag : Critical study for SIDS endpoint

17.05.2005

Type : density

Value : = .928 g/cm3 at 40 °C

Method : other:unknown

Year : GLP : no Test substance :

Source : ANGUS Chemie GmbH Ibbenbur en European Commission

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable

## 2. Physico

- Chemical Data Id 124

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Date 30.11.20 06

10 / 89 4 (documentation insufficient for assessment)

17.05.2005

2.3.1 GRANULOMETRY

2.4 VAPOUR PRESSURE

Value : = .45 hPa at 20 °C Decomposition : Method : other (measured): gas saturation method

Year : 2005

GLP : Test substance : as prescribed by 1.1

- 1.4

Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, wel l

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoint

17.05.2005 (3)

Value : = 1.33 hPa at 20 °C Decomposition : Method : other (measured): EPA Chemical Fate Test Guidel ines, CG

- 1600, "Vapor Pressure", August 1982 and Federal Register, Vol. 45, 77345, No.

772.122

- 3 (Nov. 21, 1980)

Year : 1997

GLP : yes Test substance : as prescribed by 1.1

- 1.4

Reliability : (2) valid with restrictions 2 (data from a collection)

28.06.2004 (4)

2.5 PARTITION COEFFICIENT

Partition coefficient : octanol

- water Log pow : ca.

- .63 at 20 °C pH value : Method : OECD Guide

- line 107 "Partit ion Coefficient (n

- octanol/water), Flask

shaking Method"

Year : 1996

GLP : Test substance : as prescribed by 1.1

- 1.4

Reliability : (1) valid without restriction 1 (well

- documented and acceptable for assessment) Flag : Critical study for SIDS en dpoint

17.05.2005 (5)

Partition coefficient : octanol

- water Log pow : ca.

- .74 at 20 °C pH value : Method : Year : 1999

GLP : Test substance : as pr escribed by 1.1

- 1.4

## 2. Physico

- Chemical Data Id 124

- 68

- 5

Date 30.11.20 06

11 / 89 Source : Meylan, W. (1999) SRC

- KOW for Microsoft Windows, v1.65, Octanol/Water partition coefficient estimating software.

Not GLP / Published Reliability : (2) valid with restrictions 2 (accepted calculation method)

17.05.2005

2.6.1 SOLUBILITY IN DIFFERENT MEDIA

Solubility in : Water

Value : at °C pH value : concentration : at °C Temperature effects : Examine different pol

. : pKa : at 25 °C Description : miscible

Stable : yes Deg. product : Method : Year : 1989

GLP : Test substance : as prescribed by 1.1

- 1.4

Reliability : (2) valid with restrictions 2 (data from a collection) Flag : Critical study for S IDS endpoint

10.05.2006 (6)

Solubility in : Water

Value : = 0

- 99.9 vol% at 20 °C pH value : = 11.3

concentration : .1 other:molar at 20 °C Temperature ef fects : Examine different pol. : pKa : 9.72 at 25 °C Description : miscible

Stable : Deg. product : Method : other:unknown

Year : 1989

GLP : no data

Test substance :

Source : ANGUS Chemie GmbH Ibbenburen European Commission

- European Chemi cals Bureau Ispra (VA) Reliability : (4) not assignable 4 (original reference not available)

01.03.2004

2.6.2 SURFACE TENSION

2.7 FLASH POINT

Value : ca. 78 °C Type : closed cup

Method : other: Tag Closed Cup

## 2. Physico

- Chemical Data Id 124

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Date 30.11.20 06

12 / 89 Year : 1993

GLP : Test sub stance : no data

Source : ANGUS Chemie GmbH Ibbenburen European Commission

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable 4 (documentation insufficient for assessment)

01.03.2004

2.8 AUTO FLAMMABILITY

Value : = 437.7 °C at 1013 hPa

Method : other: ASTM D 2155

Year : 1980

GLP : no Test substance :

Source : ANGUS Chemie GmbH Ibbenburen European Commission

- European Chemicals Bureau Ispra (VA) Reliability : (4) not a ssignable 4 (documentation insufficient for assessment)

01.03.2004

2.9 FLAMMABILITY

Method : Year : 1993

GLP : Test substance : as prescribed by 1.1

- 1.4

Remark : The material is considered combustible.

Source : ANGUS Chemie GmbH Ibben buren European Commission

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable 4 (documentation insufficient for assessment)

27.04.2005

2.10 EXPLOSIVE PROPERTIES

Result : not explosive

Method : Year : 1993

GLP : Test su bstance :

Remark : Based on the structure of the chemical and our experience this substance is not explosive.

Source : ANGUS Chemie GmbH Ibbenburen Reliability : (4) not assignable 4 (documentation insufficient for assessment)

01.03.2004

2.11 OXIDIZING PROPERTIES

## 2. Physico

- Chemical Data Id 124

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Date 30.11.20 06

13 / 89 Result : no oxidizing properties

Method : Year : 1993

GLP : Test substance : as prescribed by 1.1

- 1.4

Remark : Material is considered corrosive to copper, brass, and aluminum.

Source : ANGUS Chemie GmbH Ibbenburen

Reli ability : (4) not assignable 4 (documentation insufficient for assessment)

27.04.2005

2.12 DISSOCIATION CONSTANT

Acid

- base constant : 9.81

Method : other

Year : 1993

GLP : no data

Test substance : as prescribed by 1.1

- 1.4

Method : Potass ium nitrate (5.05g, 0.05 mole) was added to a 500mL volumetric flask, which was subsequently purged with N2 to exclude CO2. Approximately 490mL of purified water that had been previously boiled was added to the flask. A N2 purge ensured a good mixing of the solution. The test material and addition water were added, bringing the total volume of

the solution to 500mL. Half of the solution was transferred to a 400mL jacket beaker containing a magnetic stir bar, the pH electrode, the ATC probe, and a N2 pur ge. A Neslab headed / refrigerated circulator was used to jeep the temperature of the solution from 25.0

- 25.3C. The solution was titrated with 0.2000N hydrochloric acid and the pH and temperature was

recorded with the addition of each mL of acid.

pKa wa s calculated from the experimental data. Reliability : (2) valid with restrictions 2 (calculations based on experimental titration data)

01.03.2004 (7)

Acid

- base constant : 9.88 @ 293K

Method : other

Year : 1990

GLP : no data

Test substance : as prescribed by 1.1

- 1.4

Method : The pKa value of the conjugate acid was determined by potentiometric titration of the basic compounds at constant ionic strength. Titration curves were recorded by computerized equipment. The pH meter was read by a digital voltmeter connected to a computer interface. The motorburet was also driven by this computer interface. The sample and buffer solutions were thermostated at the required temperature before use. CO2 was excluded from the titrations using a N2 pur ge. The pKa values were calculated from titration data. The experiments were run in triplicate at each temperature, and the mean of the 3 at each temperature are reported.

Result : The triplicates were reproducible within +/

- 0.02pKa. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment)

01.03.2004 (8)

## 2. Physico

- Chemical Data Id 124

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Date 30.11.20 06

14 / 89 2.13 VISCOSITY

Test type : other Test procedure : Method : Year : 1993

GLP : Test substance :

Rema rk : Viscosity at 30C = 102 centipoise

Source : ANGUS Chemie GmbH Ibbenburen European Commission

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable 4 (documentation insufficient for assessment)

01.03.2004

2.14 ADDITIONAL REMAR KS

Memo : Henry's Law Constant

Result : 6.95 x 10^

- 3 Pa*m3 / mole

29.08.2006 (9)

## 3. Environmental Fate and Pathways Id 124

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Date 30.11.2006

15 / 89 3.1.1 PHOTODEGRADATION

Type : air Light source : Sun light Light spectrum : ca. 200

- 225 nm Relative intensit y : based on intensity of sunlight

Method : The experimental technique involved formation of OH radicals in a flow system by pulsed vacuum ultraviolet photolysis of H2O at >105nm and monitoring their resulting decay rates by detection of the OH resonance fluorescence band at 306.4nm with a cooled multiplier tube. The reaction

was studied at 50+

- 1 torr total pressure of Argon. The concentration of the amines in the argon flow was determined by UV absorption at 200

- 225nm.

Result : k1(AMP) = (2.8+

- 0.5)x10^

- 11 cm3molecule

- 1s

- 1 Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoint

01. 06.2006 (10)

3.1.2 STABILITY IN WATER

Type : abiotic t1/2 pH4 : at °C t1/2 pH7 : at °C t1/2 pH9 : at °C Deg. product : Method : Year : 2006

GLP : Test substance : as prescribed by 1.1

- 1.4

Remark : AMP lacks functional groups that would be subject to hydrolysis under typical environmental conditions. Reliability : (2) valid with restrictions 2 (dat a from a handbook or collection) Flag : Critical study for SIDS endpoint

01.06.2006 (11)

3.1.3 STABILITY IN SOIL

3.2.1 MONITORING DATA

3.2.2 FIELD STUDIES

3.3.1 TRANSPORT BETWEEN ENVIRONMENTAL COMPARTMENTS

Type : fugacity model level III

Media : other: air, water, soil sediments

Air : % (Fugacity Model Level I)

Water : % (Fugacity Model Level I)

## 3. Environmental Fate and Pathways Id 124

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Date 30.11.2006

16 / 89 Soil : % (Fugacity Model Level I)

Biota : % (Fugacity Model Level II/III)

Soil : % (Fugacity Model Level II/III)

Method : other: Mackay fugacity model

Year : 2006

Method : Input Parameters for Level III Model:

Property Value Data Temperature (°C) 25 Chemical Type 1 (can partition into all environmental compartments)

Molecular Ma ss (g/mol) 89.1 Water Solubility (g/m3) 1 x 10^6 Vapor Pressure @ 25°C (Pa) 78 Melting Point ( C) 30.5 Henry's Law Constant (Pa*m3/mole) 6.95 x 10^

- 3 Log Kow

- 0.63

Simulat ed Emission Rate (kg/hr) 1,000 (Default)

Simulated Environment Level III Default environment

Reaction Half

- lives (hr) Input to Level III Model:

Air (vapor phase) 5 Water (no susp. solids) 3,600* Soil 7,200* Sediment 7,200* Suspended Sediment 1.0 x 10^11

Fish 1.0 x 10^11

Aerosol 1.0 x 10^11

Result : Predicted Distribution to air, water, soil, and sediments

1000 kg/hr to water (most likely emission scenario) Air <0.1% (2 kg) Water 99.9% (8.38 x 10^5 kg) Soil <0.1% (144 kg) Sediment <0.1% (333 kg)

1000 kg/hr to air Air 0.5% (2770 kg) Water 59.1% (3.58 x 10^5 kg) Soil 40.5% (2.45 x 10^5 kg) Sediment <0.1% (142 kg)

1 000 kg/hr to soil Air <0.1% (278 kg) Water 58.1% (7.49 x 10^5 kg) Soil 41.8% (5.39 x 10^5 kg) Sediment <0.1% (298 kg)

1000 kg/hr simultaneously to water, air, and soil Air 0.1% (3050 kg) Water 71.2% (1.94 x 10^6 kg) Soil 28.7% (7. 84 x 10^5 kg) Sediment <0.1% (773 kg)

Conclusion : This substance has high water solubility, a low vapor pressure, and low log Kow. The substance has a low potential for adsorption to soil or sediments, and a low potential to volatilize from water or soi l to the atmosphere. If released to air, the substance will react with hydroxyl radicals. If released directly to water, the most probable emission route based on physical properties and use patterns, most of the substance will remain in the water compar tment and is expected to be biodegraded. If

## 3. Environmental Fate and Pathways Id 124

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Date 30.11.2006

17 / 89 released to soil, the substance is expected to be biodegraded. Reliability : (2) valid with restrictions 2 (accepted calculation method) Flag : Critical study for SIDS endpoint

29.08.2006 (9)

Type : f ugacity model level I

Media : other: air, water, soil sediments

Air : .1 % (Fugacity Model Level I)

Water : 99.8 % (Fugacity Model Level I)

Soil : .1 % (Fugacity Model Level I)

Biota : % (Fugacity Model Level II/III)

Soil : % (Fugacity Model L evel II/III)

Method : other: Mackay fugacity model

Year : 2006

Method : Input Parameters for Level I Model:

Property Value Data Temperature (°C) 25 Chemical Type 1 (can partition into all envir onmental compartments) Molecular Mass (g/mol) 89.1 Water Solubility (g/m3) 1 x 10^6 Vapor Pressure @ 25°C (Pa) 78 Melting Point ( C) 30.5 Henry's Law Constant (Pa*m3/mole) 6.95 x 10^

- 3 Log Kow

- 0.63 Simulated Emission Rate (kg/hr) 1,000 (Default)

Simulated Environment Level I Default environment

Result : Predicted Equilibrium Distribution Among Air, Water, Soil, and Sediments

100 000 kg total emissions

Air 0.1% (140 kg) Water 99.8% (99800 kg) Soil <0.1% (21 kg) Sediment <0.1% (<1 kg)

Conclusion : This substance has high water solubility, a low vapor pressure, and low log Kow. In the absence of advective and reactive processes, these physical

p roperties dictate that the substance will be distributed primarily in the water compartment at equilibrium. The substance has a low potential for adsorption to soil or sediments, and a low potential to volatilize from water. Reliability : (2) valid with restrictions 2 (accepted calculation method)

29.08.2006 (9)

3.3.2 DISTRIBUTION

3.4 MODE OF DEGRADATION IN ACTUAL USE

3.5 BIODEGRADATION

Type : aerobic

Inoculum : activated sludge, domestic, non

- adapted

Concentration : 2 mg/l related to Test substance

related to

## 3. Environmental Fate and Pathways Id 124

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Date 30.11.2006

18 / 89 Contact time : Degradation : ca. 40 (±) % after 28 day(s)

Result : other: not readily biodegradable Deg. product : Method : OECD Guide

- line 301 D "Ready Biodegradability: Closed Bottle Test"

Year : 1989

GLP : no Test substance : as prescribed by 1.1

- 1.4

Method : The study followed OECD Guide

- line 301 D "Ready Biodegradability:

Closed Bottle Test".

AMP was dissolved in mineral nutrient medium at a concentration of 2 mg/L. The mineral medium had been prepared from aerated de

- ionized

water amended with nutrient salts and trace element stock solutions. The reaction mixtures were inoculated with a few drops of inoculum from the discharge of a community wastewater treatment pla nt. Corresponding control mixtures were prepared with the nutrient medium; nutrient medium with inoculum; and nutrient medium with inoculum and the reference substance sodium acetate to correct for background oxygen consumption and confirm the viability o f the microbial inoculum. Equal volumes of the various reaction mixtures were dispensed into replicate bottles, sealed and incubated in the dark for 28 days at 20

- 21 oC.

The dissolved oxygen concentration in duplicate samples was measured with an oxyge n electrode on days 0, 5, 15, and 28. The percent biodegradation of the test substance was determined using the following

formula:

Dt = (mg of BODx/L) o 100 (mg of substance/L) o TOD

Dt = percent biodegradation of the test substance BODx = biological oxygen demand after x days. TOD = Theoretical oxygen consumption

Result : The results for replicate samples, expressed as percent biodegradation,

are as follows:

AMP: 5 days (0.1%, 0.1%); 15 days (1.9%, 1.9%); 28 days (40.7%, 38 .9%) Average biodegradation after 28 days was 40%.

Sodium acetate: 5 days (51.5%, 51.5%); 15 days (70.9%, 58.0%); 28 days (77.3%, 83.6%) Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoint

01.06.2006 (12)

Type : aerobic

Inoculum : activated sludge, domestic, non

- adapted

Concentration : 560 mg/l related to COD (Chemical Oxygen Demand)

related to

Contact time : Degradation : ca. 50 (±) % after 28 day(s)

Result : other: moderate degradation Deg. product : Method : other: DIN 38 412 part 25 (static test)

Year : 1986

GLP : no Test substance : as prescribed by 1.1

- 1.4

## 3. Environmental Fate and Pathways Id 124

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Date 30.11.2006

19 / 89 Source : ANGUS Chemie GmbH Ibbenburen European Commission

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable 4 (documentation insufficient for assessment)

19.05.2006

Type : aerobic

Inoculum : other: soil

Contact ti me : Degradation : ca. 50 (±) % after 35 day(s)

Result : Deg. product : Method : other: GSF

- Test

Year : 1989

GLP : no Test substance : as prescribed by 1.1

- 1.4

Source : ANGUS Chemie GmbH Ibbenburen Reliability : (4) not assignable 4 (documentation insufficient for assessment)

19.05.2006

Type : anaerobic

Inoculum : other: soil

Contact time : Degradation : ca. 15 (±) % after 35 day(s)

Result : Deg. product : Method : other: GSF

- Test

Year : 1989

GLP : no Test substance : as prescribed by 1.1

- 1.4

Source : ANGUS Chemie GmbH Ibbenburen

EUROPEAN COMMISSION

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable 4 (do cumentation insufficient for assessment)

19.05.2006

Type : anaerobic

Inoculum : activated sludge, domestic, non

- adapted

Concentration : 1 g/l related to Test substance

related to

Contact time : Degradation : ca. 4 (±) % after 5 day(s)

Result : other: low under test conditions Deg. product : Method : other: degradation in activated sludge

Year : 1989

GLP : no Test substance : as prescribed by 1.1

- 1.4

Source : ANGUS Chemie GmbH Ibbenbure n EUROPEAN COMMISSION

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable 4 (documentation insufficient for assessment)

19.05.2006

3.6 BOD5, COD OR BOD5/COD RATIO

## 3. Environmental Fate and Pathways Id 124

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20 / 89 BOD5

Method : other:unknown

Year : 1979

Concentration : 1 g/l related to Test substance

BOD5 : < 10 mg/l

GLP : no COD

Method : other: dichromate method

Year : 1979

COD : = 2050 mg/g substance

GLP : no RATIO BOD5 / COD

BOD5/COD : ca. .005

Source : ANGUS Chemie GmbH Ibbenburen Reliability : (4) not assignable 4 (documentation insufficient for assessment)

19.05.2006

3.7 BIOACCUMULATION

Species : Leuciscus idus (Fish, fresh water) Exposure period : 3 day(s) at °C

Concentration : 50 μg/l

BCF : < 1

Elimination : no data

Method : other: GSF

- test; BCF

- 3

- days

Year : 1989

GLP : no Test substance : as prescribed by 1.1

- 1.4

Method : Five Gold orfes were raised in a test vessel with 8L of water, in which the

14

- C

- labeled chemical was dissolved a t a concentration of 50ppb. The fish were not fed during the 3

- day test. The vessel was closed and the water surface was stirred with a blade stirrer gently to provide air exchange and movement of water. At the end of the experiment, the concentration o f AMP was determined by measuring the radioactivity of the fish and in the water and, from that, the accumulation factor was determined. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and

ac ceptable for assessment)

01.06.2006 (12)

Species : other: algae (Chlorella fusca) Exposure period : 1 day(s) at °C

Concentration : 50 μg/l

BCF : ca. 320

Elimination : no data

Method : other: GSF

- Test, BCF

- 1

- day

Year : 1989

GLP : no Test substance : as prescribed by 1.1

- 1.4

Source : ANGUS Chemie GmbH Ibbenburen

EUROPEAN COMMISSION

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable 4 (documentation insufficient for asses sment)

19.05.2006

## 3. Environmental Fate and Pathways Id 124

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21 / 89 3.8 ADDITIONAL REMARKS

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22 / 89 4.1 ACUTE/PROLONGED TOXICITY TO FISH

Type : static Species : Lepomis macrochirus (Fish, fresh water) Exposure period : 96 hour(s)

Unit : mg/l

NOEC : = 100 measured/nominal

LC50 : = 190 calculated

Limit test : Analytical monitoring : no Method : Year : 1980

GLP : yes Test substance : as prescribed by 1.1

- 1.4

Method : Bluegill sunfish were obtained from a commercial supplier, and held in

culture tanks with a 16

- hour light cycle, and acclimated for 14 days prior to exposure to the test material. Fish received a standard commercial fish food until 48 hours prior to testing, at which time feeding was discont inued. The fish had a mean weight of 0.21 grams and a mean standard length of

22 mm.

The static bioassay was conducted in 5

- gallon glass vessels containing 15 liters of reconstituted water (48mg NaHCO3, 30mg CaSO4, 30mg MgSO4, and 2mg KCl). Vessels we re kept in a water bath at 21

- 23C, and the fish were acclimated to the dilution water and test temperature, and fasted for 48 hours prior to testing. Dissolved oxygen was 8.8 mg/L, pH was 7.0, total hardness was 45 mg/L as CaCO3, and total alkalinity was 25 mg/L

CaCO3.

Dose levels for the definitive 96

- hour study were set in a 72

- hour range

finding test at 1, 10, 100, 560, 1000 mg/L. Dose levels for the definitive assay were 32

- 560 mg/L in logarithmic series. Concentrations are reported

based on nomin al dose.

Result : The LC50 data was reported as follows:

AMP 24

- hour LC50 = 220 (170

- 280) mg/L

AMP 48

- hour LC50 = 220 (170

- 280) mg/L

AMP 96

- hour LC50 = 190 (150

- 250) mg/L

Antimycin A 24

- hour LC50 = 0.000056 mg/L Antimycin A 48

- hour LC50 = 0.000037 mg/L Antimycin A 96

- hour LC50 = 0.000028 mg/L

There were no sublethal effects reported. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for S IDS endpoint

18.09.2006 (13)

Type : semistatic Species : Pleuronectes platessa (Fish, marine) Exposure period : 96 hour(s)

Unit : mg/l

LC50 : = 184 calculated

Limit test : no Analytical monitoring : no Method :

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23 / 89 Year : 1983

GLP : no Test substance : other TS:AMP

- 95

Result : LC50 values were calculated based on nominal concentrations and observed mortality, at a 95% CI: 24 hour LC50 = 231 mg/L 48 hour LC50 = 184 mg/L 72 hour LC50 = 184 mg/L

96 hour LC50 = 184 mg/L

Nominal temperature of the test solution was 15C (14.4

- 15C during the test). Dissolved O2 range during the test was 7.0

- 8.2 mg/L. The pH range during the test was 8.1

- 10.3. Hardness was not reported.

Test condition : In a 96

- hour, sem i

- static seawater test system, the toxicity of AMP

- 95 to Plaice was studied. Fish (20 per dose level, mean weight of 4.1g and mean length of 63.2mm) were placed in 18L glass vessels containing 10L of the test solution. The pH of the system was 8.08

- 8.14, and salinity was

34.76

- 34.82 o/oo. Concentrations tested were 100, 320, 560, 1000 mg/L. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment)

01.06.2006 (14)

Type : static Species : Leuciscus idus (Fish, fresh water) Exposure period : 48 hour(s)

Unit : mg/l

LC0 : = 320

LC50 : = 331

LC100 : = 340

Method : other: DIN 38 412 L 15

Year : 1986

GLP : Test substance : no data

Source : ANGUS Chemie GmbH Ibbenburen European Commission

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable 4 (documentation i nsufficient for assessment)

19.05.2006

4.2 ACUTE TOXICITY TO AQUATIC INVERTEBRATES

Type : static Species : Daphnia magna (Crustacea) Exposure period : 48 hour(s)

Unit : mg/l

NOEC : = 100 measured/nomi nal

EC50 : = 193 calculated Analytical monitoring : yes Method : other

Year : 1983

GLP : yes Test substance : as prescribed by 1.1

- 1.4

Method : Test procedures per "Methods of Acute Toxicity Tests with Fish, Macroinvertebrates, and Amphibians" (USEPA, 1975) were followed. The daphnia used in the test were cultured at the testing facilities, and the adult daphnia were fed a suspension of trout chow and alfalfa daily until 24 hours

## 4. Ecotoxicity Id 124

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24 / 89 prior to testing.

The static Daphnia bioassay was conducted in 250mL beakers containing 200mL of test solution. The vessels were kept at 19

- 21C, with a 16

- hour light / 8 hour dark photocycle.

An initial range

- finding study was conducted using 10 Daphnia per

concentration leve l. The range was found by beginning at 0.1 mg/L and increasing the amount of test material by a factor of 10 until a toxic level was found. Once this level had been determined, five concentrations, in duplicate, of the test compound with 10 Daphnia per b eaker were selected for their respective bioassay. These concentrations were a logarithmic series ranging from 100 to 1000 mg/L.

Remark : There was no additional data provided on the results of the range

- finding study referenced in the methods section.

Result : A computerized program calculated the LC50 for Daphnia based on the method of Stephan et.al. with a 95% CI using nominal concentrations. The

24

- hour LC50 was calculated to be 240 mg/L, and 48

- hour LC50 was calculated to be 193 mg/L.

Examinatio n of water quality parameters revealed dissolved oxygen levels ranging from 8.3 mg/L in the control at study start, and levels of 8.5 (control), 8.5 (100mg/L AMP), 8.6 (320 mg/L AMP), and 8.6 mg/L O2 (1000 mg/L AMP) at 48 hours. The pH of the control at s tudy start was 7.8. At 48 hours, the pH data collected was: 8.1 in the control, 8.3 (100mg/L AMP), 8.9 (320 mg/L AMP), and 9.4 (1000 mg/L AMP). Hardness was 225 ppm as CaCO3, and held at a temperature of 19

- 20C.

Test condition : Daphnias (Daphnia magna) first instar < 24 hours old. Test conditions were a temperature range of 19

- 20C, and a photoperiod of 16 hours light. Test solution (200mL) was placed in 250mL beakers, and ten daphnias were added per test solution. Concentrations tested were in duplic ate at 0, 100, 180, 320, 560, and 1000 mg/L AMP

- 95.

Test substance : AMP

- 95 (94.31% Active, 5.69% water, NVM 0.0004%.

Lot 216, the same as referenced in the report, "Acute Toxicity Effects of

AMP

- 95 on Bluegill Sunfish, Ninety

- Six Hour LC50" (C. Parekh, 1980).

Received as a clear liquid and stored at room temperature. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoint

01. 06.2006 (15)

Type : semistatic Species : Crangon crangon (Crustacea) Exposure period : 96 hour(s)

Unit : mg/l

EC50 : ca. 179 calculated

Limit Test : no Analytical monitoring : no data

Method : Year : 1983

GLP : no Test substance : as prescribed by 1.1

- 1.4

Method : Concentrations of AMP

- 95 tested were 0, 56, 100, 320, and 560 mg/L

under semi

- static conditions for 96 hours. The 18L glass vessel was folled with 10L of the test solution, and 20 bro wn shrimp were added to each

vessel after an on

- site acclimation period of 5 days on a specially

- prepared diet. Food was withheld during the test. The test solution was comprised of local filtered seawater (pH=8.10

- 8.16, salinity 34.86

- 34.90 o/oo), and

## 4. Ecotoxicity Id 124

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Date 30.11.2006

25 / 89 w as maintained within a temperature range of 14.7

- 15.8C. The solutions were aerated with compressed air and dissolved O2 ranged from 7.2

- 8.4 mg/L. The pH range during the test was 8.0

- 10.1. Hardness was not reported.

Remark : Detailed information regard ing pH and dissolved oxygen were not provided. Only target ranges were given.

Result : Results were as follows, with a 95% CI, based on nominal concentrations

of test material:

24

- hour LC50 = 241 mg/L

48

- hour LC50 = 179 mg/L

72

- hour LC50 = 179 mg/L

96

- ho ur LC50 = 179 mg/L The control mortality was 0% at 96 hours.

Test substance : The purity and identification of the test material is not noted. The test material was confirmed by the authors to be AMP

- 95 (2

- Amino

- 2

- methyl

- 1

- propanol). Reliability : (2) v alid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment)

01.06.2006 (16)

4.3 TOXICITY TO AQUATIC PLANTS E.G. ALGAE

Species : Scenedesmus sp. (Algae) Endpoint : growth rate Exposur e period : 72 hour(s)

Unit : mg/l

EC50 : ca. 520

Limit test : Analytical monitoring : no Method : OECD Guide

- line 201 "Algae, Growth Inhibition Test"

Year : 1989

GLP : no Test substance : as prescribed b y 1.1

- 1.4

Method : The investigations according to the guideline give false results in the case of volatile substances due to the decreasing concentration. Closing of the test vessel led to growth inhibition or death in the absence of the test

materia l. By previous passage of CO2

- enriched air and variation of the air space in the test vessel, the conditions could be optimized for the growth of the algae in the closed vessel during the 96 hour test so that the growth rate was comparable to that in an o pen vessel.

The chemical was added at increasing concentrations from a test solution prepared from a continuously illuminated algae stock culture, prepared with nutrient solution, and was placed in front of a light wall. The test vessels

were shaken sl ightly from time to time in order to prevent sedimentation of the algae. The determination of the algae concentrations was carried out, based on a calibration curve and nominal concentrations, by measuring the
extinction at time 0, 24, 48, 72, 96 hours. The inhibition was calculated with the trapezoidal formula in the Guideline. The EC50 after 72 hours was determined graphically by plotting the percent inhibition linearly against the concentration expressed logarithmically.

Result : No data other than E C50. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoint

01.06.2006 (12)

## 4. Ecotoxicity Id 124

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26 / 89 4.4 TOXICITY TO MICROORGANISMS E.G. BACTERIA

Type : aquatic Species : Pseudomonas putida (Bacteria) Exposure period : 120 hour(s)

Unit : mg/l

EC50 : ca. 101

- 202 calculated Analytical monitoring : no Method : Year : 1986

GLP : no Test subst ance : as prescribed by 1.1

- 1.4

Method : No detailed method information was provided in the report. Reliability : (4) not assignable 4 (documentation insufficient for assessment)

01.06.2006 (12)

Type : aquatic Species : other bacteria: activ ated sludge filtrate Exposure period : 120 hour(s)

Unit : mg/l

EC50 : ca. 132 Analytical monitoring : no Method : other: Sapromat

Year : 1989

GLP : no Test substance : as prescribed by 1.1

- 1.4

Source : ANGUS Chemie GmbH Ibbenburen European Commission

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable 4 (documentation insufficient for assessment)

19.05.2006

4.5.1 CHRONIC TOXICITY TO FISH

4.5.2 CHRONIC TOXICITY TO AQUATIC I NVERTEBRATES

4.6.1 TOXICITY TO SEDIMENT DWELLING ORGANISMS

4.6.2 TOXICITY TO TERRESTRIAL PLANTS

4.6.3 TOXICITY TO SOIL DWELLING ORGANISMS

4.6.4 TOX. TO OTHER NON MAMM. TERR. SPECIES

Species : other avian: Redwinged Blackbird Endpoint : mortality Exposure period : Unit : mg/kg bw

## 4. Ecotoxicity Id 124

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27 / 89 LC50 : ca. 102 measured/nominal

Method : Year : 1983

GLP : no data

Test substance :

Method : Wild

- trapped birds were preconditioned to captivity for 2

- 6 weeks and were dosed via gavage with solutions of the test chemical in propylene glycol. LD50 values were calculated by the method of Thompson (1948) and Thompson and Weil (1952).

Remark : There was no discussion of any in

- life observations or pathological findings.

Result : The calculated LD50 based on experimental results was 102 mg/kg. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoi nt

01.06.2006 (17)

Species : Musca domestica (arthropod (Diptera)) Endpoint : other: incorporation into phospholipids of larvae Exposure period : Unit : other: umol/g

Method : Year : 1965

GLP : no data

Test substance :

Method : Larvae we re grown in diets with and without choline, and growth rate and mortality were observed.

Result : When no choline is included in the diet, growth retardation is noted. The test material in the diet causes death of the larvae before the sixth day after

ha tching. If choline chloride is added to the diet with the test material, growth is normal.

Decreasing the level of choline in the diet causes an increase by 7

- 24% (of total lipids) in the proportion of phosphatidyl

- aminobutanols found in larval lipids. Reliability : (4) not assignable 4 (documentation insufficient for assessment)

26.07.2006 (18)

4.7 BIOLOGICAL EFFECTS MONITORING

4.8 BIOTRANSFORMATION AND KINETICS

4.9 ADDITIONAL REMARKS

## 5. Toxicity Id 124

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28 / 89 5.0 TOXICOKINETICS, METABOLISM AND DISTRIBUTION

I n Vitro/in vivo : In vivo Type : Metabolism Species : rat

Number of animals

Males : 42

Females : Doses

Males : The test material at a specific activity of 5uCi/umole was de livered via an IP dose of 5.1x10^6 counts/minute in 0.2mL water per rat

Females : Vehicle : water

Route of administration : i.p. Exposure time : Product type guidance : Decision on results on acute tox. tests : Adverse e ffects on prolonged exposure : Half

- lives : 1 st :
2 nd :
3 rd : Toxic behaviour : Deg. product : no Method : other

Year : 1970

GLP : no Test substance : other TS: 14C

- AMP

Method : The 42 rats were divided into 2 equal groups. One group of 5 0g Sprague

Dawley rats were given a standard rodent diet, and the other group was given a choline

- deficient diet (via a supplement of 13.2mL of ADC drops).

The test material ((3H)2

- amino

- 2

- methylpropanol) at a specific activity of

5uCi/umole was deliver ed to the rats via one intraperitoneal injection of

5.1x10^6 counts/min in 0.2mL water. The rats were sacrificed at 30 minutes, and at 1, 2, 3, 6, 24, 96 hours following dosing.

The rats were sacrificed via decapitation. Serum was stored at

- 20 and the organs and tissues were removed and immediately frozen in tubes and immersed in a dry

- ice / acetone bath. Urine and feces were collected on paper towels and the matter was extracted from the paper using a 95%

ethanol solution until no more than 5% of the initial radiation on the paper remained. The given tissues (0.5

- 1g each of the brain, heart, kidney, liver, intestine, spleen, and skeletal muscle) were homogenized with 3mL of 95% ethanol and the homogenate was centrifuged. To extract lipids from the

br ain tissue, the homogenate was was mized with a 2:1 chloroform : methanol solution, and the filtrate was stored until use. To fractionate the livers and kidneys, the mitochondria were prepared via peer

- reviewed method (Hogeboom et al.) and extracted using 95% ethanol. Microsomes were isolated by centrifugation, and the microsomes and mitochondria were washed with sucrose, then repeatedly extracted using ethanol. The particulate

- free supernatant fluid was extracted reoeatedly with ethanol

mixtures to insu re the maximum recovery possible from the cytosol. Determining the radioactivity of the serum was possible using trichloroacetic acid to make the serum protein

- free, and transferred into an ethanol solution. Aliquots of urine and tissue extracts were tra nsferred into ethanol and were similarly treated. Isolation and hydrolysis of phospholipids of mitochondria and microsomes of the liver was accomplished by removing the mitrochondrial and microsomal solvents in

## 5. Toxicity Id 124

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29 / 89 vacuo, and residues taken up in a chloroform : methanol (2:1) mixture. Aliquots from each sample were developed on a silicic acid plate and stained with I2. Stained areas were scraped from the plates, and extracted with ethanol, then chloroform:methanol. Solvents were removed, and the phospholipi d residues were hydrolyzed with HCl and stored at 105 degrees for 6hr. Petroleum ether extraction removed the fatty acids and unhydrolyzed lipids. The aqueous layer was dried, and residues were extracted in ethanol. Individual components were separated via paper chromatography, and the location of the compound was identified by staining them with I2 vapor. The papers were placed in scintillation liquid

for radioactive determinations.

Remark : Technical experts that reviewed the paper noted that there a re 2 main deficiencies in the methods employed by the authors. An IP dose, as administered in this study, gave the reviewers no data regarding the oral bioavailability of AMP, and it is assumed that accidental ingestion is one of the most likely routes of exposure to the compound. It is also noted that a lack of mass balance determination is a significant methodological deficiency, as it is impossible to determine the extent of storage of the test material, or the extent of excretion of the material. Whi le the metabolic fate of the AMP in the tissues and urine is known (AMP is stored unchanged and excreted unchanged), the relative per cent of material stored versus

excreted cannot be determined due to lack of a mass

- balance method.

The reviewers note t hat this is a reputable study with acceptable analytical and experimental methods except where noted, and is generally a useful study to use as a reference.

Result : Serum Tritium appeared in the serum 30 minutes post

- dosing. After an initial uptake of t he tritium, a large portion disappeared quickly. The radioactivity

in the serum of rats fed the choline

- deficient diet was always lower than rats on a normal diet, except at 6 hours, when the levels were about the

same.

Urine

The tritium levels in urine (cumulatively) for the normal

- diet group was greater than the choline

- deficient group up to 6 hours, at which time the levels were essentially the same. From 6

- 24 hours, the accrued radioactivity levels in the normal

- diet group exceeded that of the choli ne

deficient group. Data analyses showed that in both groups, the test material was excreted in the urine unchanged.

Tissues & Organs

Tissues from the choline

- deficient diets had a greater retention of the

amino alcohol than the normal

- diet group. The radioactivity incorporated into the mitochondria and microsomal phospholipid fractions of liver from

choline

- deficient rats increased with time, while the radioactivity in the corresponding fractions in normal rats decreased with time. The

distribution of tritium among the liver & kidney fractions was different and suggested an incorporation of the test material into various types of phospholipids. The livers from the choline

- deficient group showed a higher

level of tritium in all subcellular fractions th an the normal

- diet group at all times. Most of the tritium was found in the cytosol of both liver and kidney.

While the amount of tritium decreased with time in the mitochondrial and microsomal fractions of livers from animals fed normal diet, the amount of radioactivity increased with time in the corresponding subcellular fractions of the livers from animals receiving the choline

- deficient diet.

Test substance : The authors confirm the test substance as 2

- amino

- 2

- methylpropanol, but do not give purity o r lot. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoint

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30 / 89 01.06.2006 (19)

5.1.1 ACUTE ORAL TOXICITY

Type : LDLo

Va lue : ca. 1000

- 2000 ml/kg bw Species : rabbit

Strain : Sex : Number of animals : Vehicle : Doses : Method : Year : 1940

GLP : Test substance : no data

Method : Oral administration was ca rried out on rabbits, and the test material was delivered by stomach tube. The materials were administered in 25% w/v aqueous solutions. The solutions were neutralized in repeat experiments with HCl due to high alkalinity. Animals were fasted 24 hours prior to dosing. Following administration, animals were observed for 2

- 3 hours prior to caging, and were kept for observation more than 5 weeks after initial dosing.

Result : The lowest lethal dose was observed to be 1000

- 2000 mg/kg with both the

alkalin e and neutral solutions. Twenty to 40 minutes following dosing, animals exhibited progressive weakness to collapse, unsteadiness and incoordination ending in complete ataxia, and changes in respiration first by slowing and later by increasingly rapid rat e. No deviations in blood findings were noted, and no methemoglobin formation. There were no other findings in the post

- exposure observation period. Reliability : (4) not assignable 4 (documentation insufficient for assessment)

30.11.2006 (20)

Type : LD50

Value : ca. 2150 ml/kg bw Species : mouse Strain : Sex : Number of animals : Vehicle : Doses : Method : other: Miller and Tainter, Proc.Soc.Exper.Biol. and Med. 57:261. 1944

Year : 195 5

GLP : no Test substance : as prescribed by 1.1

- 1.4

Source : ANGUS Chemie GmbH Ibbenburen European Commission

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable 4 (documentation insufficient for assessment)

30.11.2006 (21)

Type : LD50

Value : = 2175 mg/kg bw Species : mouse Strain : Sex : Number of animals :

## 5. Toxicity Id 124

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Date 30.11.2006

31 / 89 Vehicle : Doses : 1250, 1875, 2500, 3750 mg/kg

Method : Year : 1976

GLP : no Test substance : as prescri bed by 1.1

- 1.4

Method : Mice were dosed once at dose levels of 1250, 1875, 2500, or 3750 mg/kg AMP. No further information was provided.

Result : All dose levels produced irritation, slowed activity and response to a tail pinch. All mice were grossl y normal at necropsy. Reliability : (4) not assignable 4 (documentation insufficient for assessment)

30.11.2006 (22)

Type : LD50

Value : = 2400 mg/kg bw Species : mouse Strain : Sex : female

Numbe r of animals : Vehicle : Doses : 1875, 1950, 2100, 2400, 3000 mg/kg

Method : Year : 1976

GLP : no Test substance : as prescribed by 1.1

- 1.4

Method : Mice were dosed perorally with doses of 1875, 1950, 2100, 2400, or 3000 mg/kg AMP

- 95 (5% added water). No further methods information was provided.

Result : The calculated LD50 was 2400 +/

- 89 mg/kg. Reliability : (4) not assignable 4 (documentation insufficient for assessment)

30.11.2006 (23)

Type : LD50

Value : ca. 2900 mg/kg bw Species : rat
Strain : no data

Sex : male

Number of animals : 50

Vehicle : physiol. saline

Doses : 2200, 2400, 2800, 3600, 4000 mg/kg

Method : Year : 1976

GLP : no data

Test substance : other TS: P1826 (6C 29

- 9B)

Method : The acute oral toxicity of P

- 1826 was determined using young adult male fasted rats. Five groups of 10 animals each were administered orally a single dose of P

- 1826 diluted in saline using equal volumes. Animals were observed closely fo r four hours and daily thereafter for 14 days.

Result : Doses exceeding 2800 mg/kg orally results in rapid absorbtion into the circulatory system resulting in gross damage to the liver, kidney, spleen, and respiratory system, followed by respiratory colla pse. The compound produces irritation to the stomach and duodenum at doses of 2800 mg/kg and greater.

In life observations included: traces of blood on the nostrils, labored breathing, ataxia, and death due to respiratory collapse.

## 5. Toxicity Id 124

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Date 30.11.2006

32 / 89 Necropsy findings (a nimals that died during the observation period): gas in stomach, inflamed and gaseous duodenum, liver and spleen dark, blood

clot in heart.

Necropsy findings (survivors until a scheduled necropsy): No findings in dose levels less than 2800 mg/kg. At 2800 mg/kg, 3 were grossly normal and 3 had a small amount of gas in the stomach with a slightly inflamed duodenum. At 3600 mg/kg, 1 was grossly normal and 1 was grossly normal except a stomach adhered to the liver.

The following data are reported:

LD0 = 2 200 mg/kg

LD50 = 2900 +/

- 140 mg/kg LD100 = 4000 mg/kg Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoint

01.06.2006 (24)

5.1.2 ACUTE INHALATION TOXICITY

5.1.3 ACUTE DERMAL TOXICITY

Type : LD50

Value : > 2000 mg/kg bw Species : rabbit

Strain : no data

Sex : male/female

Number of animals : 12

Vehicle : other: no vehicle

Doses : 1000, 1500, 2000 mg/kg

Method : other: Pharmacology Lab Protocol

Year : 1980

GLP : yes Test substance : as prescribed by 1.1

- 1.4

Method : Each group of rabbits was treated with 1000, 1500, or 2000 mg of test material per kg body weight (m g/kg). The desired dose was spread over the prepared abdominal skin area (abraded or smooth as designated). The skin was covered with a gauze and a sheet of impervious rubberized cloth to prevent any loss of the test material. The trunk was further encl osed with a flexible wire screen held in place by tape. The animals were returned to individual cages.

After 24 hours dermally exposed, the bindings and patches were removed, the exposed areas gently cleaned, and observed for skin irritancy. The

animals were observed for another 14 days for any gross symptoms of toxicity. At the end of the 14 day observation period, the animals were weiged, sacrificed, and the organs examined for gross pathology.

Result : At the end of the 24 hour exposure period, the intact and abraded treated skin sites were severely irritated and black in color. The sites became necrotic within two to three days and remained necrotic for the 14 days. The treated sites had severe eschar formation by the 14th day. The rabbits

in the three treatment groups lost body weight over the 14 day observation period. The animals in all treated groups showed no signs of toxicity or

## 5. Toxicity Id 124

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33 / 89 abnormal pharmacological behavior. At necropsy, all organs in all rabbits were grossly normal. The treated skin sites in all rabbits were necrotic.

Test condition : 12 Rabbits weighing 3.0 +/

- 0.5 kg were divided into 3 groups of 4 each,

and their abdomens were shaved free of hair. The skin of 2 rabbits were further prepared by abrasions. The abrasions were made 2

- 3 cm apart over the area of exposure with a blunt hypodermic needle without bleeding.

Conclusion : The acute dermal LD50 for P

- 1826 for the rabbit was >2000 mg/kg. The test material was dermally nontoxic, but was a severe skin irritant. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoint

01.06.2006 (25)

5.1.4 ACUTE TOXICITY, OTHER ROUTES

Type : other

Value : Species : rat
Strain : Sprague

- Dawley

Sex : male

Number of animals : Vehicle : water

Doses : 0.76

- 1.52 mmol : 100g body weight

Route of admin. : i.p. Exposure time : Method : Year : 1979

GLP : Test s ubstance : as prescribed by 1.1

- 1.4

Method : The test material was obtained from Aldrich Chemical Co., Milwaukee, WI. The material was used as received, no additional purification. The compound was dissolved in distilled water, and neutralized to a p H 7

- 7.2, and injected intraperitoneally. All compounds were freshly prepared, and used immediately after neutralization. The compound was tested at 0.76mmol / 100g body weight. The dosage is equivalent to that used for d

sterine in previous experiments, which always resulted in proximal tubular necrosis. Evidence of any cellular necrosis was evaluated by light microscopic examination of hemotoxylin

- and eosin

- stained sections of kidneys from animals sacrificed at 24 hours after administration of the tes t

material.

Result : 2

- Amino

- 2

- methyl

- 1

- propanol did not cause noticeable tubular necrosis

under these test conditions.

Test condition : Male Sprague

- Dawley rats (Charles River), 100

- 120g, were used. The

animals had free access to feed and water.

Relia bility : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment)

01.06.2006 (26)

Type : LC50

Value : ca. 325 mg/kg bw Species : mous e Strain : Sex : Number of animals : Vehicle : Doses : Route of admin. : i.p.

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34 / 89 Exposure time : Method : Miller and Tainter, Proc. Soc. Exper. Biol. and Med. 57:261. 1944

Year : 1955

GLP : no Test substance : as prescribed by 1.1

- 1.4

S ource : ANGUS Chemie GmbH Ibbenburen European Commission

- European Chemicals Bureau Ispra (VA) Reliability : (4) not assignable 4e (documentation insufficient for assessment)

06.03.2006

5.2.1 SKIN IRRITATION

Species : rabbit

Concentration : Exposure : Exposure time : 4 hour(s)

Number of animals : Vehicle : PDII : Result : highly irritating

Classification : irritating

Method : Year : 1940

GLP : Test substance : as prescribed by 1.1

- 1.4

Method : Animals were prepared by cl osely clipping the skin over the anterior abdominal wall. After securing the animal in the supine position, a measured amount of the test material was pipetted onto the surface of the skin. The test material was allowed to evaporate in still air, and the animal was released to the cage after the skin was dry. The treatments were repeated daily until 5 had been made.

Result : No illness resulted from skin applications of the test material, and

observations were limited to irritation. Two animals showed chemical dermititis, infiltration of the skin and subcutaneous tissues, superficial redness, and a considerable loss of hair. Burrowing lesions in the subcutaneous tissues were produced by the introduction of the test

material into small wounds on an abra ded dermal test site. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, and acceptable for assessment) Flag : Critical study for SIDS endpoint

01.06.2006 (20)

5.2.2 EYE IRRITATION

Species : rabbit

Conc entration : 100 %

Dose : Exposure time : Comment : Number of animals : Vehicle : none Result : highly irritating

Classification : risk of serious damage to eyes

Method :

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35 / 89 Year : 1975

GLP : no Test substance : as prescribed by 1.1

- 1.4

M ethod : Animals were dosed with the test material, in one study the eyes were

flushed 15

- 30 seconds after application, and in the other, they were not. No further methods information was given.

Remark : Based on known toxicity and use patterns, it is ass umed that the free amine was tested in this case and that the high pH was likely the cause of the serious eye damage noted.

Result : Unflushed eyes scored a 110 on the grading scale used, the highest possible. The test material had destroyed the vision. Flushed eyes scored 69.3/89.3 when flushed at 15 and 30 seconds, respectively. A score of 16 or higher is considered a severe irritant.

Conclusion : The test material is considered to be a severe eye irritant, and there is little

beneficial effect even w hen eyes are almost immediately flushed with

water. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoint

01.06.2006 (27)

Species : rabbit

Concentration : Dose : Exposure time : Comment : Number of animals : Vehicle : none Result : highly irritating

Classification : risk of serious damage to eyes

Method : Year : 1975

GLP : no Test substance : as prescr ibed by 1.1

- 1.4

Method : The data source does not specify other than citing Draize Techniques for

rabbits.

Remark : Based on known use patterns and toxicity of the compound, it is assumed that the serious eye damage noted in this evaluation was due to testing of the free amine at a pH~11. The high pH is likely the cause of the eye damage.

Result : Animals treated with AMP scored a 110 for eye irritation. It is considered a severe eye irritant, with a high potential for eye damage.

Conclusion : The potential for eye damage is high. Reliability : (4) not assignable 4 (documentation insufficient for assessment)

01.06.2006 (28)

5.3 SENSITIZATION

Type : Patch

- Test Species : guinea pig

Concentration : 1 st : Induction 5 %

2 nd : Challenge 2. 5 %

3 rd : Challenge 5 %

Number of animals : 30

Vehicle : water

Result : not sensitizing

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36 / 89 Classification : not sensitizing

Method : other: Buehler

Year : 1982

GLP : no Test substance : as prescribed by 1.1

- 1.4

Method : One group was treated with 0.5mL of 10% P

- 1826 solution, a negative control group was treated with saline, and a positive control group was treated with dinitrochlorobenzene (DNCB solubilized in alcohol and made to volume with saline). After 24 hours, the patches were removed, and sites were cleaned and scored at 24 and 48 hours for erythema and edema according to Draize (Draize, JH, "Appraisal of the Safety of Chemicals in Foods, Drugs, and Cosmetics". Assoc. of Food and Drug Officials of the United States, p. 48, 1957). At 48 hours, the application was repeated with each group, and continued 2

- 3 times per week until 10 applications were made. Animals were allowed a 2 week recovery period, and then challenged at a virgin site. The test and negative control animals were

chal lenged with 0.5mL of 2.5% and 5% solutions of P

- 1826. Positive and negative control animals were also challenged with 0.03% DNCB solution. After 24 hours, the test material was cleaned away, depilated, and three hours later scored for erythema and edema. Sites were scored again at 48

hours.

Test material is considered a sensitizer if the challenge elicits skin reactions in a large number of test animals when compared to the negative

control.

Result : During the induction, the 10% P

- 1826 solution was found to be mildly irritating to all animals in the test group, so the remaining 8 doses during the induction were made with a 5% solution. The positive control, DCNB, elicited a mild to strong reaction during the 10 applications. There was one

death in the positive control group, but was deemed not treatment

- related via necropsy.

At challenge with 2.5% and 5% solutions of P

- 1826, none of the animals in the test or negative control groups showed any skin reactions at 24 hours, but the positive control an imals showed mild skin reactions at 48 hours.

Nine of ten positive controls when challenged with DNCB at 24 hours showed skin reactions, and 7/10 showed reactions at 48 hours. Only 4/10 negative controls showed reactions when challenged with DNCB at 24

h ours, and none at 48 hours.

Test condition : Thirty male guinea pigs (250

- 300g each) were divided into 3 groups of 10 each. The animals' backs and flanks were shaved free of hair. The guinea pigs were topically treated with the solutions applied under a n occlusive patch.

Conclusion : 2

- Amino

- 2

- methyl

- 1

- propanol was a non

- sensitizer in the topical sensitization test in guinea pigs, under these test conditions. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoint

18.09.2006 (29)

Type : Intracutaneus test Species : guinea pig

Concentration : 1 st : Induction .1 %

2 nd : Challenge .05 %

3 rd : Challenge .01 %

Number of animals : 30

Vehicle : water

Result : not sensitizing

Classification : not sensitizing

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37 / 89 Method : Year : 1982

GLP : no Test substance : as prescribed by 1.1

- 1.4

Method : One group was treated with 0.05mL of 1% P

- 1826 solution, a negative

contr ol group was treated with saline, and a positive control group was treated with dinitrochlorobenzene (DNCB solubilized in alcohol and made to volume with saline). After 24 hours, sites were cleaned and scored for erythema and edema according to Draize (Dr aize, JH, "Appraisal of the Safety of Chemicals in Foods, Drugs, and Cosmetics". Assoc. of Food and Drug Officials of the United States, p. 48, 1957). At 48 hours, the application was repeated with each group, and continued 2

- 3 times per week until 10 ap plications were made. Animals were allowed a 2 week recovery period, and then challenged at a virgin site. The test and negative control animals were challenged with 0.1mL of 0.05% and 0.01% solutions

of P

- 1826. Positive and negative control animals wer e also challenged with 0.3% and 0.03% DNCB solution. After 24 hours, they were depilated, and three hours later scored for erythema and edema. Sites were scored again at 48 hours.

Test material is considered a sensitizer if the challenge elicits skin reactions in a large number of test animals when compared to the negative

control.

Result : During the induction phase, the first injection at 1% and second injection at

0.5% P

- 1826 induced necrotic lesions, so the remaining 8 injections were

made with 0. 1% solutions. The 0.3% DNCB sites were necrotic for the entire 10 injections.

At challenge with 0.05% and 0.01% P

- 1826, one animal in the test group showed mild reactions with 0.05%, but none of the negative controls challenged with P

- 1826 showed any rea ctions at 24 or 48 hours. In the repeat challenge, none of the animals in the test group showed any

reactions with P

- 1826, but of the negative control group, 4 animals at

0.05% and 1 animal at 0.01% showed skin reactions at 24 hours.

A challenge with the positive control induced skin reactions in the positive control group. The 0.03% solution did not elicit any skin reaction at 48

hours.

Test condition : Thirty male guinea pigs (250

- 300g each) were divided into 3 groups of 10

each. The animals' backs a nd flanks were shaved free of hair. The guinea pigs were intradermally

- injected with the solutions. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Cri tical study for SIDS endpoint

01.06.2006 (30)

5.4 REPEATED DOSE TOXICITY

Type : Sub

- chronic Species : rat
Sex : male/female

Strain : other: CD

Route of admin. : gavage Exposure period : 13 weeks Frequency of treatm. : 5 days / week Post expo sure period : Doses : 0, 500, 750, 1100, 1700 mg/kg at pH of 7 & 11+ Control group : yes, concurrent vehicle

## 5. Toxicity Id 124

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38 / 89 LOAEL : < 500 mg/kg

Method : Year : 1977

GLP : no Test substance : other TS: P

- 1826 (assumed to be AMP

- Regular)

Method : Dose Preparation The dose solution for pH=11+ was prepared by adding the neat test material to distilled water in order to achieve a concentration adequate to

dose the animals 1

- 5mL of dose solution according to the animals' bod y weights and dose group. The dose solution of pH=7 was prepared by adding the neat test material to distilled water, neutralizing the solution to pH=7. Because the resulting stock solution is purple in color, the solution was decolored using activated c harcoal. The stock solution was then diluted as appropriate to obtain solutions adequate to dose the animals

with 1

- 5mL solution according to their body weights and the targeted dose.

Dose Administration The solutions were administered 5 days per week fo r 13 weeks, once per day by oral gavage. The dose was adjusted to the body weight of the rat by administration of different volumes of solution (1

- 5mL).

In

- life Observations & Measures Animals were observed 5 days/week for pharmacological effects of the test material. The observations were general, including signs of altered food and water consumption, irritability, sedation, hair and cutaneous alterations, external inflammation, hyperventilation, convulsions, neuromuscular paralysis, and altered activit y. Body weights were recorded weekly, and dead animals were recorded and autopsied as soon as

discovered.

Hematology & Clinical Chemistry Measures The following blood parameters were measured: hemaglobin, packed cell colume (hematocrit), erythrocyte coun t, leucocyte count, differential count, transaminases, glutamic

- oxalacetic and glutamic

- pyruvic, total protein, blood urea nitrogen (BUN), Sodium, Potassium, Chloride, and Creatinine. For urine, Ph, protein, occult blood, glucose, bilirubin, sediment, an d specific gravity were measured.

Pathology Only gross pathology was reported.

Result : Mortality A high mortality rate was recorded for animals with the pH=11 doses. No

treatment

- related mortality was noted for the pH=7 groups.

Behavior and Appearance General observations on the pH=11 groups included hyperventilation and respiratory difficulty, neither of which could be confirmed at necropsy. Non

treatment related pneumonia was noted in some rats. Rats generally showed hyperirritability and hyperacti vity at the time of AMP administration. In the pH=11 rats receiving more than 750mg/kg, abdominal swelling, nasal and mouth bleeding were noted the first few days of the study. The abdominal swelling was always followed by death. Some loss of hair

aroun d the mouth and face was noted for pH=11 rats, and assumed to be

treatment

- related. General observations for rats receiving the dose at pH=7 were hyperventilation (lesser extent than pH=11 rats), and limited hyperactivity and hyperirritability. Some of th e rats also showed loss of hair around the mouth and face. In a few rats, the effect was accompanied by skin ulceration due likely to excessive salivation, or a local irritancy by the compound.

Body Weight, Food Consumption Trends

## 5. Toxicity Id 124

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39 / 89 There was no significan t difference in the rate of body weight change between all dose groups relative to the controls. Food and water consumption were qualitatively noted. Food consumption appeared
normal, while AMP

- administered rats appeared to drink more frequently

than con trols.

Hematology & Clinical Chemistry Neither the pH=7 or pH=11 females showed significant differences in hematocrit, hemoglobin, erythorcyte counts, or leukocytes. The males from the pH=11 group in the 1100 mg/kg dose group showed a significant

decreas e in hematocrit, hemoglobin, and red blood cell count due to a loss

of blood.

pH 11 group males Group HCT HgB RBC(x10^6) WBC Control (10) 44+

- 1 16.5+

- 0.4 7.41+

- 0.29 8.2+

- 1.2 500mg/kg (8) 43+

- 1 16.7+

- 0.9 6 .20+

- 0.47 13.6+

- 2.6 750mg/kg (7) 43+

- 1 16.2+

- 0.5 6.31+

- 0.34 9.4+

- 2.5 1100mg/kg(2) 40+

- 1 14.9+

- 0.3 5.94+

- 0.10 9.0+

- 1.6 1700mg/kg (none surviving)

A slight but significant decrease is seen in the RBC of rats receiving 500

and 75 0 mg/kg AMP. There was no significant effect on hematocrit at these exposure levels. The pH=7 group (only at 1700mg/kg) showed significant decreases in hematocrit and hemoglobin.

pH 7 group Group HCT HgB Control (10) 46+

- 1 17.1+

- 0.1 500mg/kg (9) 43+

- 2 16.4+

- 0.5 750mg/kg (10) 42+

- 1 16.4+

- 0.6 1100mg/kg(10) 42+

- 1 16.4+

- 0.8 1700mg/kg (9) 40+

- 1 15.1+

- 0.2

No significant differences were measured in Na, K, Cl, Ca, total serum protein , BUN, or creatinine for either pH group at any dose level. SGPT values were within normal ranges for all dose groups except for pH=7 females at 1700mg/kg, and pH=7 males at 1000 and 1700 mg/kg, where the values were significantly elevated.

Urinalysis

Ur inalysis was performed only for the pH=11 groups. The data indicated some proteinuria.

Gross Pathology

There were no treatment

- related grossly

- recognizable changes observed at

necropsy. Test condition : CD rats were received from Charles River weighing 90

- 100g, and were acclimated a minimum of one week to the laboratory prior to the experimental start. The rats were divided into groups of 10 per sex per dose level, for each of 2 pH's. They were housed in groups of 2

- 3 per plastic cage, and fed via over head racks. Food and water was provided ad libitum and animals were housed in rooms with controlled temperature, humidity, and photocycles.

Conclusion : LOAEL and NOAEL cannot be established because the authors do not specifically state, or provide detai led data regarding the lowest dose level

at which in

- life observations (respiratory difficulty, hyperirritability, hyperactivity, etc.) were seen. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- document ed, and acceptable for assessment) Flag : Critical study for SIDS endpoint

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40 / 8 9 01.06.2006 (31)

Type : Sub

- chronic Species : dog

Sex : male/female

Strain : Beagle

Route of admin. : oral feed Exposure period : 90 days Frequency of treatm. : continuo us Post exposure period : Doses : 0, 25, 600, 2500 ppm Control group : yes, concurrent vehicle

NOAEL : = 25 ppm LOAEL : = 600 ppm Method : Year : 1981

GLP : no Test substance : other TS: AMP

- 95/HCl 48.6% ai. Lot 109

- 1

Method : The AMP was administered orally via the diet as AMP

- 95/HCl to 32 Beagle dogs (4/sex/dose group) at dose levels of 0, 25, 600, and 2500 ppm for a three month period. The control animals received an untreated diet. Body weights were measured pretest, weekly thereafter, and after fasting at termination; food consumption was measured daily and presented at weekly intervals. Feed samples of each dose level were assayed for homogeneity, stab ility, and verification of dose concentration. Results were presented in a subsequent report from the performing laboratory, and not presented in this report. Animals were observed for mortality and gross signs of toxicologic or pharmacologic effects twi ce daily, and dogs were given a detailed physical examination pre

- test and weekly thereafter. Ophthalmoscopic examinations were performed pretest and at termination. Hematology, clinical chemistry, and urinalysis parameters were analyzed twice pre

- test, and Month 1 and 3. Blood was obtained via jugular venipuncture. Urine was collected in stainless steel metabolism pans attached to each animal's permanent cage. Dogs were fasted overnight prior to blood collections, and were not dosed until after sample s were collected. Hematology parameters evaluated included: hemoglobin, hematocrit, erythrocytes, platelets, clotting time, prothrombin time, and total and differential leukocytes. Clinical Chemistry parameters included: serum glutamic oxaloacetic transa minase, serum glutamic pyruvic transaminase, alkaline phosphatase, lactic acid dehydrogenase, blood urea nitrogen, fasting glucose, cholesterol, total protein, albumin, globulin, A/G ratio, total bilirubin, direct bilirubin, sodiom, potassium, chloride, an d calcium. Urine was evaluated for the following: Gross appearance, specific gravity, pH, protein, glucose, ketones, bilirubin, occult blood, urobilinogen, and microscopic analysis.

On days 96

- 99, animals were necropsied and tisses were preserved for

we ights (brain, ovaries, pituitary, thyroid, heart, adrenals, spleen, testes, kidneys, liver) and microscopic evaluation (the above plus aorta, bone, sternum, epididimus, esophagus, eye, colon, duodenum, ileum, lungs, mesenteric lymph node, mammary gland, sc iatic nerve, pancreas, parathyroids, prostate, salivary gland, skeletal muscle, skin, spinal cord, stomach, thymus, thyroid, trachea, urinary bladder, and gross lesions).

Statistical analyses were performed on data collected for body weight, food

consu mption, hematology, clinical chemistry, organ weights, and organ:body weight ratios. Mean values of all dose groups were compared to control at each time interval (where appropriate). Statistically significant

differences from the controls are indicated.

Species and strain of test animals, route and method of test substance

## 5. Toxicity Id 124

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41 / 89 administration, and dose levels were established by the sponsor. The study was performed at Bio/dynamics, INC.

Remark : Results section references Appendix tables and data, but none is included in the Dow Chemical copy of the report. Test material intake, therefore,

cannot be estimated.

Result : Mortality: All animals survived the duration of the study.

Physical Observations Included emesis, loose stool, alopecia, and slight penil e or vaginal discharge. These findings occurred sporadically in the control and/or treated animals and are not considered by the authors to be treatment

related.

Ophthalmology Pretest and terminal examinations revealed no significant ocular changes

throu gh the study. There is no evidence of compound

- induced ocular damage. Body Weight: No statistically

- significant differences were found in any dose group when compared to animals of the same sex in the control group. There was a 10% mean body weight reduc tion noted in the high dose group relative to the control for males and females in latter phases of the study, suggesting to the authors a slight compound

- related effect.

Food Consumption

There was no treatment

- related effect of the test material on food consumption at any dose level.

Hematology Pretest hematology values were within the normal physiological range for all animals. There were no statistically

- significant differences between the control and treated animals at Month 1 testing. At Month 3, t here was a trend toward reduced prothrombin time observed in the treated males and females with statistically significant differences noted in the top two dose levels for males only (p<0.01). In addition, a slight trend torward reduced hemoglobin concentr ations was found in the treated females. Only the mean value for the high dose, which was slightly below the normal physiological range, was statistically significant (p<0.05). No other biologically significant differences were observed in any other hema tology parameter evaluated.

Clinical Chemistry Pretest mean clinical chemistry values were within the normal physiological range for males and females. At both the Month 1 & 3 evaluations, statistically significant elevations in serum glutamic oxaloaceti c transaminase, serum glutamic pyruvic transaminase, and alkaline phosphatase activities occurred in the high dose males and females. The magnitude of the increases ranged from 2

- 4 fold for serum glutamic
oxaloacetic transaminas, 12

- 28 fold for serum glut amic pyruvic
transaminase, and 5

- 6 fold for alkaline phosphatase. These significant increases were limited to the high dose animals only, and were similar at months 1 & 3 for both sexes. Other findings included reduced mean

cholesterol values in both sex es at the above intervals, while direct bilirubin concentrations were slightly higher in the treated animals. No other toxicologically significant differences occurred in the other clinical chemistry parameters evaluated. The above effects observed in th e high dose animals suggest a high degree of hepatocellular damage in the high dose animals with no significant changes in the low or mid dose groups. The combination of increased serum glutamic oxaloacetic transaminase and serum glutamic pyruvic transami nase levels, both of which are present in high concentrations in hepatocytes, are indicative of hepatocellular necrosis. The increased alkaline phosphatase and direct bilirubin levels

## 5. Toxicity Id 124

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42 / 89 are suggestive of cholestasis and/or bile duct epithelial necrosis. It appears, therefore, that the liver is the promary target organ, based on the clinical chemistry data, when the test substance is administered via the

diet.

Urinalysis There is no evidence of a compound

- related effect on any of the urinalysis
parameters s tudied.

Organ Weight/ Organ:Body Weight Ratios A trend towards increased liver and/or liver/body weight rations was ovserved in both the mid and high dose male and female dogs. These differences were slight but tend to support the conclusion that the liv er is the principle site of toxic action. Also, mean kidney weights and kidney/body weight ratios were increased in the high dose males, and in mid and high dose females. Other organ weight and organ/body weight differences were marginal and appear unrel ated to the administration of the

test material.

Pathology Two females and one male from the high dose group were found to have tan discoloration and mottling in liver upon gross postmortem evaluations. No other such findings were present in any of the o ther treated or control animals. Other gross findings occurred sporadically or were present in the treated and control animals to a similar degree. Microscopic findings included vacuolization, periportal cirrhosis, and bile duct hyperplasia in the

liver sections in all high

- dose males and females. Lipid deposition within the cellular vacuoles was confirmed by staining. The periportal cirrhosis was characterized by hepatocellular necrosis and fibrosis. These findings were more severe in the high

- dose fe males as compared to the males. In addition, one low dose female had liver findings similar to the above, while

2 mid

- dose males exhibited minimal fatty changes in the liver. Other microscopic findings occurred sporadically in the treated and/or control

animals and were not related to the administration of the test material. The above microscopic findings correlate well with the clinical chemistry data, as both indicate hepatotoxicity induced by the test material in the high dose animals. The findings n oted in the liver of the one low dose female, in the absense of altered clinical chemistry data, as well as the absence of similar microscopic findings in the mid dose group, are considered to be of uncertain significance (this may represent a response to treatment in an unusually sensitive animal, or may be a spontaneous finding).

Test condition : Study was run with Beagle dogs, 4/sex/dose. They were 4.5

- 5 months of age at arrival to the Bio/dynamics laboratory, and 5.5

- 6 months of age at

the initiation of the study. Weight range at the start of treatment was 7.7

- 9.6kg for males (mean 8.7kg), and was 6.3

- 9.7 for females (mean 8.1kg). The dogs were immunized previously against distemper, hepatitis, leptospirosis, and rabies by the supplier. Fecal examin ations were conducted on all dogs by the performing lab during the one month acclimation period. Baseline clinical laboratory tests were also performed. They were housed in elevated metal grid cages, and fed 400g of Purina Canine Meal Diet (#5007) prese nted for 4.5 hours. Fresh food was presented daily. Water was provided ad libitum by an automated water system. The dogs were maintained on a 12

- hour photocycle, and temperature was monitored twice daily.

Dogs were ranked by body weight and distributed into 4 groups of 4/sex so that body weights in each group were comparable. Groups were assigned to control and dose levels randomly. Each dog was assigned a unique identification number on an ear tag and on the animal's cage. In addition, each dog had an ear tatoo bearing the USDA number.

Test substance : AMP

- 95/HCl (48.6% AI). Lot #109

- 1 (11/14/80) provided by International Minerals & Chemical Corporation (the study sponsor).

## 5. Toxicity Id 124

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43 / 89 Conclusion : The results of this study agree with those from a concurrentl y

- run study (Bio/dynamics #80

- 2503), utilizing the same test material and similar dose levels in rats, which also indicated the liver is the principle target organ from dietary administration of the test material. The degree of changes in the present stud y clearly indicates that dogs are more sensitive to the hepatotoxic effects of the test material. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critic al study for SIDS endpoint

01.06.2006 (32)

Type : Chronic Species : dog

Sex : male/female

Strain : Beagle

Route of admin. : oral feed Exposure period : 1 year Frequency of treatm. : continuous Post exposure period : Doses : 0, 1.1, 11, 110 p pm Control group : yes, concurrent vehicle

NOAEL : > 110 ppm Method : Year : 1990

GLP : no data

Test substance : other TS: AMP

- HCl (47.1% AMP)

Method : Animals were dosed via the diet, which was tested by the supplier (Purina) for contaminants. The test article was incorporated into the diet on a weight/weight basis using a premix. Fresh food was prepared with the test article weekly. The animals were offered 400g of the diets daily. Any

uneaten food w as weighed and recorded. Drinking water was provided throughout the study ad libitum. Six males and six females were assigned to each of 4 dose groups (0, 1.1, 11, 110 ppm). Daily observations were made of the general appearance, behavior, the

presence o f any signs of toxicity or pharmacologic effects and mortality. Body weights were recorded prior to dosing, and weekly thereafter. The fasting terminal body weight was also recorded. While weighing, the animals were examined for lesions or other signs o f toxicity. Each animal had an ophthalmoscopic examination once pre

- study, weeks 1, month 3,6,9,12 by a veterinarian. Blood samples were obtained pre

- study, and at 3, 6, 9, 12 months via venipuncture of fasted animals. The following parameters were recor ded: hematocrit, hemoglobin, erythrocyte count, leukocyte count, platelet count, prothrombin time, MCH, MCHC, and MCV, Calcium, Phosphorous, Chloride, Sodium, Potassium, glucose, serum alkaline phosphatase, serum aspartate aminotransferase, serum analine a minotransferase, gamma glutamyl, transpeptidase, blood urea nitrogen, total protein, albumin, globulin, creatinine, bilirubin, and serum protein. Urine was examined for pH, specific gravity, volume and appearance, protein, glucose, acetone
bodies, creatin ine, and microscopic sediment.

Post

- mortem examinations included a complete gross examination of the external surfaces, all orifices, the cranial cavity, the external surface of the brain, the thoracic, pelvic, and abdominal cavities and their viscera, cer vical tissues and organs, and the carcass. Two animals per sex per dose were sacrificed at 6 months, with the remainder at 12 months. Organ weights were recorded for the liver, kidneys, testes, thyroids, adrenals, and brain.

Tissues that were removed an d preserved are the same as current OECD guidelines.

Experimental dates: 5/19/1988 to 7/25/1989

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44 / 89 Remark : Test material intake is estimated based on week 36 body weights and food consumption.

Males estimated (mg/kg/day)

Control 0 1.1ppm .031 11ppm 0.31 110ppm 2.98

Females

estimated (mg/kg/day)

Control 0 1.1ppm .029 11ppm 0.31 110ppm 2.55

Result : There were no general in

- life observations made for any treated or control animal. Likewise, there were no ophthalmoscopic observations noted for any animal. Neither an ANOVA or a Kruskal

- Wallis non

- parametric ANOVA revealed any statistical significance in food consumption patterns for any dose level in either sex. There was no statistically significant differences in body weights for any of the animals throughout the study. Clinical chemistry f indings for males revealed a slight decrease in the low

and mid

- dose groups' albumin

- globulin ratio. Since there was no dose

response (the high dose group was not different than the control), it was judged to be a spurious finding and not treatment

- relate d. High

- dose

males at 9 and 12 months showed differences in serum albumin via serum electrophoresis, but was not corroborated by actual clinical chemistry measurements, and was not considered by the authors to be biologically

significant. Throughout the study, findings for females included differences in serum sodium, serum ALT, and serum AST, but were transient, did not appear dose

- related, and were judged by the authors to be not related to

the administration of AMP. There was no effect of AMP administ ration on hematology, and likewise the urinalysis revealed no treatment

- related effects. There were no effects on organ weights or organ/body weight ratios that were attributed to test material administration by the authors. At necropsy, evaluation of the tissues revealed several isolated observations, none of which were attributed to the administration of the test material. There were no neoplastic observations in any of the dogs sacrificed at either 6 months or one year of administration in their diets.

Test condition : Twenty nine male and 29 female beagle dogs were received from Laboratory Research Enterprises, Kalamazoo, MI, and 25 of the healthiest of each sex were used for study. The animals were selected based on criteria of the US FDA (1982). T he dogs were 4 months of age at arrival to the testing laboratory. Animals were identified by an assigned identification number on a collar, housed singly for an acclimation period, and were examined by a laboratory veterinarian for general health status. Animals were stratified by body weight, and assigned randomly to their treatment

groups. Test substance : 2

- amino

- 2

- methyl

- 1

- propanol supplied by Angus Chemical Co, Northbrook, IL. Supplied as an aqueous solution of AMP

- HCl. Concentration of AMP in

th e solution was 47.1%.

Conclusion : Based on the findings under these study conditions, there is no effect at any dose level on general appearance, behavior, body weight, food consumption, ophthalmoscopic exams, clinical chemistry, hematology, organ weight s, or tissue histopathology. Based on the absence of statistically and biologically significant findings in dose

- response patterns,

the No

- Observed Effect Level for AMP in the diets of Beagle dogs in greater than 110 ppm. Reliability : (2) valid with res trictions

## 5. Toxicity Id 124

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45 / 89 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment) Flag : Critical study for SIDS endpoint

05.06.2006 (33)

Type : Sub

- acute Species : rat
Sex : male/female

Strain : Sprague

- Dawley

Route of admin. : oral feed Exposure period : 8 weeks Frequency of treatm. : continuous Post exposure period : Doses : 0, 1000, 2000, 4000, 8000, 16000 ppm Control group : yes, concurrent vehicle

LOAEL : ca. 1000 ppm Method : Year : 1976

GLP : Test substance : other TS: P

- 1826 (assumed to be AMP

- regular)

Method : The test material was fed in the diet at dosage levels of 1000, 2000, 4000, 8000, 16000 ppm. Ten male and 10 female rats were used at each dos e level and also in a control group. The compound was mixed with distilled water so that the same total volume of liquid was mixed with the food for each group. The compound

- water premix was mixed with a small amount of diet, and the food premix was mixe d with the total diet. Control rats also

received distilled water in their diet at the same volume as the treated diets.

Rats were observed daily for changes in general behavior and appearance. Individual body weights and sex group food consumption wer e recorded weekly.

At the termination of the study, all surviving rats at the 8000ppm and 16000ppm doses were necropsied. In addition, 2 males and 2 females at each of the other dose levels plus control were likewise necropsied. Livers plus any gross le sions from all animals necropsied were paraffin embedded, sectioned, and stained and examined microscopically.

Remark : Estimated test material intake on week 4, based on mean body weights and food consumption:

MALES Dose Group Material Intake (m g/kg/day)

Control 0 1000ppm 244 2000ppm 474 4000ppm 912 8000ppm 1901 16000ppm 4698

FEMALES Dose Group Material Intake (mg/kg/day)

Control 0 1000ppm 436 2000ppm 806 4000ppm 1628 8000ppm 3424 16000ppm 7579

Result : In

- life observations made beginning week 3 included emaciation, rough
hair coat, and scattered 1 x1mm lesions over the dorsal surfaces of a few rats in the 16000ppm group. After week 3, the incidences increased so

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46 / 89 that all rats in the high dose showed similar clinical signs. Hair loss on the dorsal surface was also noted in rats in this group beginn ing at week 5 and continuing thereafter. One female rat at the high dose showed general paleness and tremors prior to death at week 2. Two female rats in the 16000ppm group died during the study.

Rats in the 16000ppm group gained slightly less weight as compared to control rats. Gains in the other dose groups were similar to the control. Food consumption patterns were similar for all animals.

At necropsy, all 16000ppm surviving rats, and all rats at the 8000ppm dose level were necropsied. Two males and two females from each of the other groups were also examined. Alopecia and focal skin erosions and ulcerations found in rats at the 16000ppm dose level were viewed as possibly compound related. Hepatocyte vacuolation descrived in rats from

all the t reated groups, with increasing severity at the higher dose levels, was considered compound related. Microscopic skin lesions found at the 16000ppm dose level, like the corresponding gross lesions, were viewed as possibly compound related.

Test condition : Sixty male (133

- 183g at study start) and female (115

- 146g at study start) CD rats were received from Charles River Labs, and were housed individually in hanging canges, maintained in a temperature and humidity

controlled room. Purina rodent chow and wat er were provided to the

animals ad libitum.

Conclusion : LOAEL = 1000ppm LOAEL under these test conditions was 1000ppm. Observed effects at 1000 ppm included alopecia and focal skin erosions. Reliability : (2) valid with restrictions 2 (meets general ly accepted scientific standards, well

- documented, and acceptable for assessment)

01.06.2006 (34)

Type : Sub

- acute Species : mouse Sex : male/female

Strain : CD

- 1

Route of admin. : oral feed Exposure period : 8 weeks Frequency of treatm. : cont inuous Post exposure period : Doses : 0, 200, 400, 800, 1600, 3200 ppm Control group : yes, concurrent vehicle

NOAEL : > 3200 ppm Method : Year : 1976

GLP : Test substance : other TS: P

- 1826 (assumed t o be AMP

- regular)

Method : The test material was fed in the diet at dosage levels of 200, 400, 600, 800, 1600, 3200 ppm. Ten male and 10 female mice were used at each dose level and also in a control group. The compound was mixed with distilled

water s o that the same total volume of liquid was mixed with the food for each group. The compound

- water premix was mixed with a small amount of diet, and the food premix was mixed with the total diet. Control mice

also received distilled water in their diet at the same volume as the treated

diets.

Mice were observed daily for changes in general behavior and appearance. Individual body weights and sex group food consumption were recorded weekly.

At the termination of the study, all surviving mice at the 3200 ppm dose

## 5. Toxicity Id 124

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47 / 89 were necropsied. In addition, 2 males and 2 females at each of the other dose levels plus control were likewise necropsied. Livers plus any gross lesions from all animals necropsied were paraffin embedded, sectioned,

and stained and examined mic roscopically.

Remark : Estimated test material intake for the animals were calculated based on 4 week body weight and food consumption:

MALES Estimated mg/kg/day

Control 0 200ppm 1.244 400ppm 2.088 800ppm 3.360 1600ppm 8.950 3200ppm 14.380

FEMALES Estimated mg/kg/day

Control 0 200ppm 1.600 400ppm 3.146 800ppm 6.070 1600ppm 12.870 3200ppm 27.742

Result : No changes cons idered to be related to compound were seen in the general behavior and appearance. There was no treatment

- related mortality. Increases in body weight over the course of the study were comparable for control and all treated animals. Food consumption valu es were also similar for the control and treated animals.

There were no compound

- related gross lesions observed in any of the necropsied mice. There were likewise no microscopic lesions that were

considered treatment

- related in the livers of any mice nec ropsied.

Test condition : Sixty male (20

- 31g at study start) and female (18

- 27g at study start) CD

- 1 mice were received from Charles River Labs, and were housed individually in hanging canges, maintained in a temperature and humidity controlled

room. Pur ina rodent chow and water were provided to the animals ad

libitum.

Conclusion : The NOAEL for CD

- 1 mice under these conditions is >3200ppm. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, an d acceptable for assessment)

01.06.2006 (35)

Type : Sub

- acute Species : dog

Sex : male/female

Strain : Beagle

Route of admin. : oral feed Exposure period : 4 weeks Frequency of treatm. : continuous Post exposure period : Doses : 0, 600, 1800 , 5400, 16200 ppm Control group : yes, concurrent vehicle

NOAEL : ca. 600 ppm Method : Year : 1976

GLP : Test substance : other TS: P

- 1826 (lot 6C29

- 9B), purity not given, supplied by the study

sponsor

## 5. Toxicity Id 124

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48 / 89 M ethod : P

- 1826 was administered in the diet at doses 600, 1800, 5400, and 16,200 ppm. One male and one female was assigned to each dose level. The compound was mixed with distilled water and a small amount of chow to create a premix. The premix was used to mix the total diets at each dose level. Dogs were observed daily for changes in general behavior and appearance. Individual body weights and food consumption were recorded weekly. Blood and urine was collected pre

- study and at week 4. Hematology st udies included hemoglobin, hematocrit, total erythrocyte count, total and differential leukocyte counts and erythrocyte sedimentation rate. Clinical chemistry parameters included: fasting glucose, urea nitrogen, serum glutamic oxalacetic and pyruvic trans aminase activities, serum alkaline phosphatase activity, total protein and albumin. Albumin to globulin ratios were calculated. Urinalysis included description of color and appearance, measurement of volume, pH, specific gravity, occult blood,

and micros copic examination of the sediment.

Remark : Estimated test material intake was calculated based on week 2 body weights and food consumption data:

Males

Control estimated (mg/kg/day) 600ppm 21.9 1800ppm 59.6 5400ppm 25.37 (food consumption extremely low) 16200ppm (1 gram of food consumption reported)

FEMALES

Control estimated (mg/kg/day) 600ppm 19.0 1800ppm 70.86 5400ppm 198.0 16200ppm 168.8 (food consumption low)

Result : Soft stool and/or diarrhea frequently were noted for the female dog at 5400ppm, and for both dogs at 1800 and 16200 ppm. A dry nose and

mouth were noted the second week of s tudy for both dogs at the 16200 ppm. There was no mortality during the study. Marked losses in body weight were seen for the male dog at the 5400ppm dose level and for both dogs at the 1800 and 16200 ppm dose levels. All other dogs maintained or

showed gains in body weight. A marked decrease in food consumption was noted for the male dog at the 5400ppm dose level, and almost total anorexia was noted for both dogs at the 16200ppm dose level. Slightly elevated hemoglobin, hematocrit, and erythrocyte coun t were noted for the female high

- dose female at 4 weeks. Also at 4 weeks, a slight neutropenia for the male dog at the 5400 and 16200 ppm dose levels was noted. No other changes were seen in hematological studies. At the 600ppm dose level, the female sh owed a slight increase in SGPT activity. Moderate to marked increases were seen in alkaline phosphatase and SGPT activity for all dogs at the 1800, 5400, and 16200ppm dose levels. Slight to moderate increases in SGOT activity also were seen for all dogs at the 5400 and 16200ppm dose levels. No other changes were seen in clinical chemistry parameters. There were no unusual values seen in urinalyses. There were no compound related gross pathologic lesions observed at necropsy in any dog. There appeared to be a compound related decrease in absolute liver weights at 5400 and 16200 ppm and several dogs also had decreases in relative liver weights as compared to values from untreated dogs of similar age and weight. Microscopically, all dogs except the male at 600ppm had evidence of toxic liver injury, characterized by hepatocyte vacuolation, necrosis of scattered individual hepatocytes and accumulations of a greenish

- brown pigment in Kuppfer cells. Sinusoids in

## 5. Toxicity Id 124

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49 / 89 the centrolobular areas of some livers appeare d dilated, suggesting that atrophy of some liver parenchymal tissue had occurred. Decreased liver weight noted in some dogs would be consistent with atrophy of hepatic parenchymal tissue; with only one dog of each sex, the authors had difficulty assessing such. Microscopic lesions in tissues other than the liver were those which commonly occur spontaneously in dogs and were not considered significant.

Test condition : Four male and four female beagle dogs (6.9

- 10 kg) were housed individually in metal met abolism cages, maintained in a temperature and humidity controlled room, and offered Purina dog chow and water ad

libitum.

Test substance : P

- 1826 (lot 6C29

- 9B) purity not given, supplied by the study sponsor Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment)

01.06.2006 (36)

Type : Sub

- acute Species : rat
Sex : male/female

Strain : Long

- Evans

Route of admin. : gavage Exposure period : 5 days Frequency o f treatm. : once per day Post exposure period : Doses : 0, 500, 1000, 2500 mg/kg Control group : yes NOAEL : ca. 500

- 1000 mg/kg

LOAEL : ca. 1000

- 2500 mg/kg

Method : Year : 1977

GLP : Test substance : as prescribed by 1.1

- 1.4

Method : Four groups of rats were gavaged daily for 5 days with AMP in distilled water at dose levels of 0, 500, 1000, 2500 mg/kg. These were

administered from stock sol utions at a rate of 1mL/100g body weight. The pH's of the dosing solutions all exceeded 11. The rats were observed daily and body weights were taken through the 5 days of dosing and up to the 15th day of the study. The survivors were sacrificed and necro psied on the 15th day. Tissues were taken for further study. Body weights and organ weights were measured for livers and kidneys. Representative samples of livers and kidneys were selected for microscopic examination.

Result : Mortality

All five males at 2500mg/kg were dead by test day 3. Two females died by test day 11 at 1000mg/kg, and all five females at 2500mg/kg were dead by test day 3.

Bodyweight There were slight decreases in bodyweight noted for surviving rats in all dose groups, relative to t he controls. There were no statistically significant findings.

Necropsy All livers of the surviving rats looked normal grossly. Upon further examination of tissues, there was no data presented in the report that suggest a harmful effect of the compound u nder these experimental conditions. The gross and microscopic findings represent the usual profile of spontaneous diseases that occur in rats. There were no significant findings in kidney and liver organ weights or organ:body weight ratios for any dose l evel relative to the controls.

Source : Albany Medical College, unpublished data. FOI# F86

- 38214.

## 5. Toxicity Id 124

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50 / 89 Test condition : Forty Long

- Evans rats (90

- 100g), 5/sex/dose, were acclimated to the lab for more than one week. Rat chow and water were provided ad libit um. Conclusion : In general, no dose

- related pathological effects were seen in the livers of rats. The mortality of the higher dose animals appeared to be associated with GI tract effects. The clinical chemistry studies show significant changes in some of the parameters, and cannot be disregarded. There exists, however, a strong possibility that the clinical chemical differences as well as GI tract pathology is associated with the strong alkalinity (pH>11) of the dose preparation, rather than an effect of the AMP alone. Under conditions of this study, the following LOAEL and NOAEL are noted:

Males LOAEL = 2500 mg/kg NOAEL = 1000 mg/kg

Females LOAEL = 1000 mg/kg NOAEL = 500 mg/kg Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment)

19.05.2006 (37)

Type : Sub

- acute Species : monkey

Sex : female

Strain : other: Rhesus

Route of admin. : drinking water Exposure period : 5 days Frequency of treatm. : once d aily Post exposure period : Doses : 500, 1000 mg/kg Control group : no Method : Year : 1977

GLP : no data

Test substance : as prescribed by 1.1

- 1.4

Method : The monkeys were dosed with AMP in distilled water at dose levels of 500 and 1000 mg/k g. Dosing continued for 5 days or until the higher dose monkey died. Body weights were taken daily. The low dose monkey was observed for 14 days prior to necropsy on the 15th day, and the high dose monkey was necropsied at the time of death. Hematology and clinical chemistry parameters were evaluated, and animals were examined at necropsy for gross and microscopic findings.

Result : The high dose female died 2 hours after the 3rd dose. Cause of death was recorded as gastrointestinal hemorrhaging. Bot h monkeys showed an approximately 10% decrease in body weight from the first to last

measurement.

Hematology results showed no significant changes in RBC, HgB, or Hct and only a slight increase in WBC counts. In the surviving monkey, significant differen ces in Ca++, BUN, Creatinine, SGPT, CPK, and OCT

were seen.

The liver in both the low and high dose animals, upon necropsy evaluation, were found to be grossly and microscopically within normal limits.

Source : Albany Medical College, unpublished data. FOI #F86

- 38214.

Test condition : Two Rhesus monkeys were acclimated to the conducting laboratory for over one week. Baseline values for clinical chemistry, hematology, and urinalysis were performed prior to dosing.

Conclusion : In general, no dose

- relat ed pathological effects were seen in the livers of

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51 / 89 monkeys. The mortality of the high dose animal appeared to be associated with GI tract effects. The clinical chemistry studies show significant changes in some of the parameters, and cannot be disregarde d. There exists, however, a strong possibility that the clinical chemical differences as well as GI tract pathology is associated with the strong alkalinity (pH>11) of the dose preparation, rather than an effect of the AMP alone. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment)

19.05.2006 (37)

Type : Sub

- acute Species : mouse Sex : male

Strain : Swiss Webster

Route of admin. : inhalation Exposure perio d : 180 minutes Frequency of treatm. : once Post exposure period : 20 minute recovery in room air

Doses : 185

- 1160 mg/m3 Control group : yes Method : Year : 1996

GLP : no data

Test substance : as prescribed by 1.1

- 1.4

Method : The method of ae rosol generation for AMP is not specified in detail. The animals were placed in plethysmographs attached to the generation apparatus such that their heads were extended into the interior of the

chamber. A latex collar served as a seal around the neck of the mouse and held its head in place. Pressure transducers permitted the measurement of pressure changes created by mice during inhalation and exhalation. Relative tidal volume changes and respiratory frequency were measured. Each experiment consisted o f a 20 minute control period in air alone, exposed to air (sham exposed), or exposed to the test material for

180 minutes. A 20

- minute recovery period was given immediately following exposure. Animals were observed for 1 week for any mortality.

Using th e breathing patterns measured during the experiment by the plethysmograph, sensory irritation and pulmonary irritation can be

identified.

Remark : The article failed to cite detailed atmosphere generation methods. The atmosphere concentration is reported only as a range of 185

- 1160 mg/m3;

the absence of a time

- weighted average concentration makes interpretation of the results difficult, as the actual dose to the animal is not known. The analysis of the atmosphere by Miran IR does not preclude the possibi lity aerosol exposure at a higher concentration than reported.

Result : In general, exposure to the test material resulted in immediate sensory irritation, and persisted throughout the 3 hour exposures. The test material also produced pulmonary irritatio n, but the effect was not immediate (2

- 3 hour delay in onset). Animals exposed to AMP failed to return to control levels during the 20 minute recovery period following exposure, thus recovery was considered poor. The test material is considered moderate in potency relative to other test materials tested concurrently under these test

conditions.

Test substance : Supplied to the conducting lab by the United Auto Workers and General Motors Corporation National Joint Committee om Health and Safety.

Conclusi on : AMP may be considered a sensory and pulmonary irritant at the concentration range tested in mice under these study conditions. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and
accepta ble for assessment)

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52 / 89 01.06.2006 (38)

Type : Sub

- chronic Species : rat
Sex : male/female

Strain : Sprague

- Dawley

Route of admin. : oral feed Exposure period : 3 months Frequency of treatm. : Post exposure period : Doses : 0, 25, 150, 250, 250 0 ppm in feed Control group : yes, concurrent vehicle

LOAEL : = 2500 ppm Method : Year : 1981

GLP : no Test substance : other TS: AMP

- 95/HCl 66.1ai (Lot 109

- 1)

Method : Appropriate amounts of test materi al were mixed with standard lab rodent diet weekly, to provide 0, 25, 150, 250, or 2500 ppm to the animals. The animals received the test material continuously in the diet for 90

- 92 days prior to necropsy. Homogeneity of the test material in the feed was assayed by sampling all dose levels (from the top, middle, and bottom of the containers). Samples from all dose levels were assayed for stability in the feed at day 0, 3, 4, 7, 10, and 14. Concentration of the test material in the feed was analyzed for all dose levels on test weeks 1, 2, 3, 4, 6, 8, 12,

and 13.

Animals were observed twice daily for mortality and gross signs of toxicologic or pharmacologic effects. A detailed physical examination for signs of local or systemic toxicity, pharmacologic ef fects, and palpable masses was performed weekly. Ophthalmoscopic examinations were performed pretest and at the end of month 1 and 3. Body weights were recorded twice pretest, weekly during treatment, and terminally after fasting. Food consumption was r ecorded weekly beginning the week prior to study start, and test material was calculated from food consumption data

based on nominal concentration of the test material in the feed.

Blood was analyzed for the following parameters (pretest, month 1 & 3):

he moglobin, hematocrit, erythrocytes, platelets, total & differential leukocytes, erythrocyte morphology, serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, alkaline phosphatase, blood urea nitrogen, fasting glucose, total protein, albumin, globulin, A/G ratio, cholesterol, sodium, potassium, calcium, total bilirubin, and lactic acid dehydrogenase.

Urine was analyzed at month 1 and 3, and gross appearance, specific gravity, pH, protein, glucose, ketones, bilirubin, occult blood, u robilinogen, and microscopic analysis were evaluated.

Animals dying spontaneously received a gross postmortem exam. All others surviving to necropsy were euthanized under anesthesia, underwent a gross postmortem exam, and select tissues were weighed and/ or preserved for examination. Organ weight was collected for adrenals, brain, gonads, heart, kidneys, liver, pituitary, and spleen and they were also preserved for examination. In addition, the following were preserved:
aorta, blood smear, bone and bone marrow, epididymus, eye, cecum, ileum, duodenum, jejunum, lungs, mesenteric & mediastinal lymph nodes, mammary glands, sciatic nerve, pancreas, parathyroid, prostate, salivary gland, seminal vesicle, skeletal muscle, skin, spinal cord, stomach, thymus, thy roid, trachea, bladder, uterus, gross lesions, and tissue masses.

## 5. Toxicity Id 124

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53 / 89 Statistical analysis included body weight, food consumption, hematology, and clinical chemistry parameters, organ weights and organ:body weight ratios. Mean values of all dose groups were compared to control at each time interval where appropriate. Statistically significant differences from control are indicated in appendices.

Remark : Results & methods sections referenced appendix data, which is not provided in Dow's copy of the report. As such, test material intake (mg/kg/day) could not be estimated.

Result : Mortality All animals survived the test material administration, with the exception of 2

females

- one died following Month 1 blood collection, and the other was

sacrificed on test day 18 due to poor food consumption, constipation, lethargy, and swollen abdomen. The condition was not considered due to

the test material.

Physical Observations All observations noted were interpreted by the author to be related to a viral

infection (sialdacryoadenitis) in all dose groups at approximately the same frequency, primarily in males except where noted, and included: alopecia, swollen salivary glands (both sexes), opacities, excessive lactimation, and chromodacryorrhea.

Ophthalmoscopic Ex aminations
Pretest exams revealed no notable abnormalities. At months 1 & 3, a significant increase in a variety of lesions, which appeared to be infectious in origin, were noted. The observations were likely secondary to the sialodacryoadenitis infectio n which can lead to opacity and other ocular lesions. The incidence of the abnormalities was similar in all dose groups and both sexes, and therefore does not appear to be treatment

- related.

Body Weight Body weight differences between the treated and con trol animals of both sexes were slight during the course of the study. No consistent pattern

related to the administration of the test substance was evident. It is concluded by the authors that the dose levels selected did not affect body weights signifi cantly.

Food Consumption Among treated males, slight reductions in food consumption were evident during weeks 1

- 3 as compared to the control. Thereafter, mean food consumption values were comparable in all groups. Mean food consumption values tended to be slightly higher in the treated females as compared to the controls during the course of the study. The increases were slight and are not considered to be of toxicological significance.

Test Substance Intake

Test substance intake values based on nomina l dietary concentrations were approximately proportional to the dietary concentrations. Test substance intakes were highest for all groups during the early phase of the study and gradually declined reaching a plateau by approximately week 10.

Since the f emales consumed more feed on a body weight basis, their test substance intake calues were highter as compared to the males within a given group.

Hematology Pretest hematology values were within normal limits for both sexes. Occasional statistically signi ficant differences were noted in the group mean hematology data for the treated animals as compared to the control groups, for both males and females. However, as all means were generally within the normal physiological range and no consistent

treatment

- r elated pattern evident, it was concluded by the authors that the administration of the test material did not significantly alter the hematology

## 5. Toxicity Id 124

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54 / 89 parameters evaluated.

Clinical Chemistry
Pretest values were normal for both sexes. At month 1, total bilirubi n values were significantly decreased in the 2 highest dose groups while total protein and globulin levels were significantly increased in only the highest dose group. Among the females, month 1 alkaline phosphatase was slightly elevated in all but the co ntrol and low dose groups. Statistical significance was noted for only the 250ppm group. At Month 3, a trend toward increased SGOT levels was found among the treated males, while BUN levels tended to be lower in dosed animals as compared to the

controls. In addition, LDH was significantly increased in the high dose only. Among the females, SGOT, SGPT, and LDH were significantly elevated in the high dose only. In addition, total protein and globulin values were significantly decreased in the high dose. Other significant differences were
noted in both sexes at the 1 and 3 month evaluations. However, these differences were generally slight and the values within the normal physiological range for animals of this species and strain. The combination

of inc reases in SGOT, SGPT, and LDH in the males and decreased BUN in the females at Month 3 suggest that the liver may be a target organ. The magnitude of these differences was slight and generally confined to the high dose group. Therefore any alterations in liver function would be expected to be minimal in degree.

Urinalysis There were no effects on urinalysis parameters noted in any animal over the course of the study.

Organ Weight and Organ:Body weight ratios Among the males, the high dose kidney/body w eight ratio was increased, while the absolute liver weights were decreased in the 150 and 250 ppm groups. Among the females, the absolute and relative liver and spleen weights were significantly increased in the 25 and 2500 ppm groups, respectively. The changes were slight and appear due to biological

variation as no dose

- response pattern emerged. The possible exception is the tendency toward increased liver weights in the treated females. As previously discussed, the clinical biochemistry values sugges t the liver as a target organ for the test material. Frequently, liver weight changes are secondary to biochemical altherations in liver morphology and/or function.

Pathology Gross post

- mortem examinations revealed no significant compound

- related

abnorma lities. Lesions noted occurred sporadically and/or were found in both the control and treated animals at approximately the same incidence. Microscopic evaluations revealed patchy hepatocellular vacoulization in the

livers of 3/20 males and 4/20 females i n the high dose group This finding ranged from nomimal to moderate in degree. These vacuoles, presumably containing lipid, suggest that the liver may be a target organ for the test compound, as the clinical chemistry evaluations showed increased serum

c oncentrations of hepatic enzymes.

Test condition : Male and female Sprague

- Dawley rats were received from Charles River Breeding Laboratories and were 4 weeks of age at arrival to the Bio/dynamics Lab. They were allowed to acclimate to the laboratory for a minimum of 14 days, and were housed individually in elevated stainless steel cages. They were fed Purina Certified Lab Chow (#5002) ad libitum, and fresh food was presented weekly. Water was provided ad libitum via an automated water system. Photocy cle was 12 hours light per 24 hour cycle, and temperature was monitored twice daily. They were approximately 7.5 weeks of age at the start of treatment. Animals were given a pre

- assignment physical examination and unsuitable animals were

excluded from as signment to test groups. The animals were assigned randomly to groups (20/sex/dose), and were identified with an numeric

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55 / 89 metal ear tag. Reliability : (3) invalid 3 (methodological deficiencies, high incidence of viral infection in test animals)

01.06 .2006 (39)

5.5 GENETIC TOXICITY ‘IN VITRO‘

Type : Bacterial reverse mutation assay System of testing : S. typhimurium (tester strains TA98, TA100, TA1535, TA1537) and E.coli (tester strain WP2 uvrA)

Test concentration : 100, 333, 1000, 3333, 5000 ug per plate Cycotoxic concentr. : Metabolic activation : with and without

Result : negative

Method : OECD Guide

- line 471

Year : 1996

GLP : yes Test substance : as prescribed by 1.1

- 1.4

Method : Positive controls, appropriate to the given teste r strains, were used as

follows:
All strains, activated

- 2

- aminoanthracene
WP2uvrA, nonactivated

- methyl methanesulfonate
TA98, nonactivated

- 2

- nitrofluorene
TA100, TA1535, nonactivated

- sodium azide
TA1537, nonactivated

- 9

- aminoacridine

The concentr ation varied according to positive control compound. The sterility of the test article was verified.

Preliminary Toxicity Assay Ten dose levels of the test article were plated, one plate per dose, with overnight cultures of TA100 and WP2 uvrA on selectiv e minimal agar in the presence and absence of rat liver S9 activation.

Mutagenicity Assay An initial and confirmatory assay was used to evaluate the mutagenic potential of the test material. A minimum of 5 dose levels of the test article,

with vehicle an d positive controls, were plated with tester strains in the presence and absence of rat liver S9 activation. All dose levels of test article, vehicle controls, and positive controls were plated in triplicate. The test systems were exposed to the test art icle via the plate incorporation methodology described by Ames, et.al.( (1975) and updated by Maron and Ames (1983). Plates were coded according to the testing laboratory's standard procedures. Plates were incubated for 48

- 72 hours at 35

- 39C.

Plates not counted immediately following the incubation period were stored

at 2

- 6C until the colonies could be counted.

Toxicity and degree of precipation were scored relative to the vehicle control plate using a well

- defined 9

- point scale. Colonies were counted

e ither by an automated colony counter or by hand unless the assay was preliminary or the plate exhibited toxicity. Plates exhibiting test article precipitate that interferes with an automated colony counter were counted by hand.

Mean and standard deviatio n of the number of revertants per plate were calculated and reported. For a positive finding, the test material must

cause a dose

- related increase in mean revertants per plate of at least one tester strain with a minimum of 2 increasing concentrations of test article.

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56 / 89 Positive finding for TA1535 and TA1537 is an increase in mean revertants at the peak of dose

- response greater than, or equal to, 3x mean control value. Positive finding for TA98, TA100, WP2 uvrA is an increase in mean revertants at the peak of dose

- response greater than, or equal to, 2x mean

control value.

Remark : GLP's were followed except for the following exceptions: 1. The identity, strength, purity, and composition or other characteristics to

define the test or control article were n ot determined by the testing facility. 2. Analyses to determine the uniformity, concentration, or stability of the test or control mixtures were not performed by the testing facility. 3. The stability of the test or control article under the test conditi ons was not determined by the testing facility.

Result : Water was selected as the solvent of choice. The test material was soluble in water at approximately 100ug.mL, the maximum concentration tested.

Preliminary Toxicity Assay

The maximum dose teste d was 5000ug/plate, achieved by using a concentration of 100mg/mL and a 50uL plating aliquot. Based on the findings, the maximum dose plated in the mutagenicity assay was 5000ug/plate. Neither precipitate nor appreciable toxicity was observed.

Mutagenic ity Assay Neither precipitate nor appreciable toxicity was observed in the initial or confirmatory assays. In neither the initial nor confirmatory assay was there a positive response with any of the tester strains at any dose level, in the presence or abs ence of S9 activation. The mean of each positive control

exhibited at least a 3

- fold increase in the number of revertants over the mean value of the respective vehicle control, indicating a valid test.

Test condition : Salmonella tester strains were reci eved directly from Dr. Bruce Ames, University of California, Berkeley, and the E.coli was received from the National Collection of Industrial and Marine Bacteria, Aberdeen, Scotland.

Tester strains TA98 and TA1537 are reverted from histidine dependence (a uxotrophy) to histidine independence (prototrophy) by frameshift mutagens. Tester TA1535 is reverted by mutagens that cause basepair substitutions. Tester TA100 is reverted by mutagens that cause both frameshift and basepair substitution mutations. Spec ificity of the reversion mechanism in E.coli is sensitive to basepair substitution mutations, rather

than frameshift mutations.

To assure that cultures were harvested in late log phase, the length of

incubation was controlled and monitored. Each culture wasmonitored spectrophotometrically for turbidity and was harvested at a percent transmittance yielding a titer of approximately 10^9 cells per milliliter.

Acoclor 1254

- induced rat liver S9 was used for the metabolic activation system. The S9 was prepare d from male Sprague

- Dawley rats induced with a single IP injection of Acroclor 1254, 500mg/kg, five days prior to sacrifice. Each batch was assayed for its ability to metabolize.

Conclusion : The results of this study indicate that, under the conditions of this study, the test material did not cause a positive response with any of the tester strains in the presence and absence of Acrclor

- induced rat liver S9. Reliability : (1) valid without restriction 1 (guideline, GLP study) Flag : Critical study f or SIDS endpoint

01.06.2006 (40)

Type : Mammalian cell gene mutation assay System of testing : L5178Y/TK+

- Mouse Lymphoma

Test concentration : 0.5, 1.5, 5.0, 15, 50, 150, 500, 1500, 5000 ug/mL Cycotoxic concentr. : Metabolic activation : with an d without

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57 / 89 Result : negative

Method : OECD Guide

- line 476

Year : 1996

GLP : yes Test substance : other TS: AMP

- 95

Method : Aroclor 1254

- induced rat liver S9 was used as the metabolic activation
system.

A preliminary toxicity assay was used to esta blish the optimal dose levels for the mutagenesis assay. L5178Y cells were exposed to the vehicle alone and to concentrations of the test article ranging from 0.5

- 5000 uL in both the absence and presence of S9

- activation. Cell population density

was dete rmined at 24 and 48 hours after exposure to the test article. Toxicity was measured as suspension of growth relative to the growth of the solvent controls. The mutagenesis assay, initial and independent repeat, was used to evaluate the mutagenic potenti al of the test article. L5178Y mouse lymphoma cells were exposed to the vehicle alone and 10 concentrations of the test article in both the absence and presence of S9. Positive controls, with and without S9, were tested concurrently.

Tubes containing L5 178Y/TK+

- cells with F0P medium or S9 activation
mixture, and 100uL of test solution at concentrations of 0.5, 1.5, 5.0, 15, 50, 150, 500, 1500, 5000 ug/mL were prepared. Positive controls treated with MMS and 7,12

- DMBA were also prepared. Treatment tube s were gassed with CO2 and incubated in the dark for 4 hours. Cells were washed twice, resuspended, gassed with CO2 in air again, and placed on a roller. Cells were counted, and numbers were adjusted if necessary. For expression of TK+

- cells, cells wer e placed in a cloning meduim of granulated agar, mixed with Trifluorothymidine (TFT) or viable count, prewarmed and filled with cloning media. The tubes were centrifuged, supernatant decanted, and cells resuspended in cloning media. Tests were

conducted i n duplicate. TFT or Viable Count were added to the flask and placed on a shaker. The suspension was placed in petri dishes, placed in cold storage for 30 minutes, then incubated for 10

- 14 days.

Diameters and numbers of colonies were recorded, and analyze d relative to positive and solvent controls. Previous published data indicates that the large colony mutants received localized damage, likely in the form of a point mutation or small deletion within the TK locus, and small colony mutants received damage to collateral loci concordant with the loss of TK activity.

Remark : Study followed GLP's except for the following: 1. The identity, strength, purity, and composition or other characteristics to define the test or control article were not determined by t he testing facility. 2. Analyses to determine the uniformity, concentration, or stability of the test or control mixtures were not performed by the testing facility. 3. The stability of the test or control article under the test conditions was

not determ ined by the testing facility.

Result : Sterile distilled water was the solvent of choice. The test material was miscible in water at 500 mg/mL, the maximum tested concentration.

Preliminary Toxicity Assay The high dose tested was 5000ug/mL. Osmolality o f the solvent control was 300mmol/kg and of the highest dose was 396mmol/kg. Suspension growth relative to the solvent control was 13% at 5000ug/mL without S9 activation, and 31% with S9 activation. Based on these results, the dose

levels chosen for the mutagenesis assay ranged from 500

- 5000ug/mL for

both the non

- activated and S9

- activated cultures.

Mutagenesis Assay

In the non

- activated system, cultures treated with 1000

- 500ug/mL were

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58 / 89 cloned and produced a range in suspension growth of 11

- 55%. In the S 9

- activated system, cultures treated with 500

- 5000ug/mL were cloned and produced a range in suspension growth of 9

- 98%. No treated cultures exhibited mutant frequencies more than 55 mutants per

10^6 clonable cells over the solvent control. A dose

- response trend was

not observed in either the activated or non

- activated systems. Over the concentration ranges, total growth for the non

- activated cultures ranged

from 12

- 60%, and 10

- 137% for the S9

- activated cultures. Results of the independent repeat assay ar e similar (12

- 62% and 32

- 124% for non

activated and S9

- activated system suspension growth, respectively; 14

- 67% and 29

- 191% for non

- activated and S9

- activated system total growths, respectively). There was likewise no increase in mutant frequencies over t he solvent control. Colony sizing for the positive control yielded the expected increase in small colonies, thereby verifying the methods used to detect small colony mutants.

Conclusion : The results of the L5178Y/TK+

- Mouse Lymphoma Mutagenesis Assay

in dicate that, under the conditions of this study, 2

- amino

- 2

- methyl

- 1

- propanol did not cause a positive response in the non

- activated and S9

- activated systems, and was concluded to be negative. Reliability : (1) valid without restriction 1 (guideline, GL P study) Flag : Critical study for SIDS endpoint

27.07.2006 (41)

Type : Ames test System of testing : S.cerevisiae (D

- 4) and S.typhimurium (TA98, TA100, TA1535, TA1537, and TA1538)

Test concentration : 0.01, 0.1, 1.0, 5.0 microliters per plate Cy cotoxic concentr. : Metabolic activation : with and without

Result : negative

Method : Year : 1976

GLP : no Test substance : other TS:Referred to as P

- 1826 (AMP) Lot #6029

- 9B. No purity information was included in the final report.

Method : P

- 18 26 was evaluated for its potential mutagenic effect using the microbial procedure of Ames, et.al. Saccharomyces cerevisiae D

- 4 and five strains of Salmonella typhimurium (TA98, TA100, TA1535, TA1537, and TA1538)

were used for the evaluation in which the reaction mixture was tested with and without rat liver cell 9000Xg supernate metabolic activation.

Male rats were euthanized and the organs were immediately dissected and placed in cold sucrose buffered solution with TRIS at a pH of 7.4. The

organs were washed twice with the buffered solution and homogenized. The homogenate was centrifuged, the supernatant was extracted, and retained in cold storage, and were used for the activation study.

Approximately 10^9 cells from a log phase culture of each indica tor strain were added to test tubes containing 2mL of molten agar supplemented

biotin and histidine. For non

- activation tests, the four dose levels of the test compound were added to the contents of the appropriate tubes and poured over the surfaces of se lected agar plates. In activation tests, the tissue supernatant and required cofactors were added to the overlay tubes. Four dose levels of the test chemical were added to the appropriate tubes, which were then mixed and the contents poured over the surf ace of a minimal agar plate and allowed to solidify. Plates were incubated for 48 to 72 hours at 37C, and scored for colonies. Positive (nonactivated assay methylnitrosoguanidine, 2

- nitrofluorene, quinacrine mustard; activated assay 2

- anthramine, 2

- acety laminofluorene, 8

- aminoquinoline, dimethylnitrosamine) and solvent controls (water, DMSO, or saline

## 5. Toxicity Id 124

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59 / 89 depending on the positive control used) were run with each assay.

The numbers of colonies on each plate were counted and recorded. The positive and the so lvent controls are provided as reference points.

Result : The test compound, P

- 1826, was examined for mutagenic activity in a series of in vitro microbial assays employing Salmonella and Saccharomyces indicator organisms. The compound was tested directly and in the presence of liver microsomal enzyme preparations from Aroclor

induced rats. The appropriate positive controls for each system and strain elicited a strongly

- positive result in all cases, validating the assay.

The compound was tested over a se ries of concentrations such that there was either quantitative or qualitative evidence of some chemically

- induced physiological effects at the high dose level. The low dose in all cases was below a concentration that demonstrated any toxic effect. The do se range employed for the evaluation of the compound was from 0.01uL to 5uL per plate.

The results of the tests conducted on the compound in the absence of a metabolic system were all negative.

The results of the tests conducted on the compound in the pr esence of the rat liver activation system were all negative as well.

Test substance : Referred to as P

- 1826 (AMP) Lot #6029

- 9B. No purity information was included in the final report.

Conclusion : The test compound did not demonstrate mutagenic activity in any of the assays conducted in this evaluation and was considered not mutagenic

under these test conditions. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment)

01.06.2006 (42)

5.6 GENETIC TOXICITY ‘IN VIVO‘

Type : Micronucleus assay Species : mouse Sex : male/female

Strain : ICR

Route of admin. : i.p. Exposure period : 1 ip injection

Doses : vehicle control, 16, 30, 60 mg/kg, and positive control C P (50mg/kg)

Result : negative

Method : OECD Guide

- line 474 "Genetic Toxicology: Micronucleus Test"

Year : 1998

GLP : yes Test substance : other TS: AMP

- 95 (lot 35662)

Method : In each the Pilot & Toxicity Studies (designed to set appropriate dose

le vels for the micronucleus assay), and the Micronucleus Assay, mice were weighed immediately prior to dose administration and the dose volume was based on individual body weights. They were observed after dosing, and daily thereafter for 3 days for clinica l signs of chemical effects. Body weights were recorded prior to dose administration and 1 and 3 days after dosing. Mice were given a single IP injection of the test article in water, the vehicle alone, or the positive control substance.

Pilot Study: 2 male mice were dosed with 1, 10, 100, or 1000 mg test material /kg body weight, and 5 males and 5 females were dosed with 2000 mg/kg.

## 5. Toxicity Id 124

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60 / 89 Toxicity Study: 5 male and 5 female mice were dosed with 200, 400, 600, 800 mg/kg.

Based on the results of the pilot a nd toxicity studies, the high dose for the micronucleus test was set at 60 mg/kg, which was estimated to be approximately 80% of the LD50/3.

Micronucleus Assay: Male and female mice were dosed with the positive control (5/sex/dose), vehicle control (5/sex /dose), 16 mg/kg (5/sex/dose), 30 mg/kg (5/sex/dose), or 60 mg/kg (5/sex/dose with an additional 5/sex/dose in the event of mortality) via a single IP injection of the test material or the positive control (cyclophosphamide). At the scheduled sacrifice ti mes (24 and 48 hours), up to 5 mice / sex / dose were sacrificed, the femurs exposed, and bone marrow aspirated into a syringe of fetal bovine serum. The bone marrow cells were pelleted by centrifugation and the supernatant drawn off. The

cells were resu spended and a small amount of suspension spread onto a glass slide. Two to four slides were prepared per animal. The slides were fixed in methanol, stained, and permanently mounted. Slides were coded and scored blindly. Using oil immersion, 2000 polych romatic erythrocytes were scored for the presence of micronuclei. The number of micronucleated normochromatic erythrocytes in the field of 2000 polychromatic erythrocytes was enumerated. The proportion of polychromatic erythrocytes to total erythrocytes was also recorded per 1000 erythrocytes. Statistical significance was determined using the

Kastenbaum

- Bowman tables which are based on the binomial distribution.

The test article was considered to induce a positive response if a dose

responsive increa se in mocronucleated polychromatic erythrocytes was observed and one or more doses were statistically

- elevated relative to the vehicle control (p>0.05) at any sampling time. If a single dose group was significantly elevated without a dose

- response trend, the assay was considered a suspect or unconfirmed positive and a repeat assay recommended. The test article would be considered negative if no statistically significant increase in micronucleated polychromatic erythrocytes above the concurrent vehicle con trol was observed at any sampling time.

Remark : Exceptions to GLP's included: 1. The identity, strength, purity, and composition or other characteristics to define the test or control article were not determined by the testing facility. 2. Analyses to determine the uniformity, concentration, or stability of the test or control mixtures were not performed by the testing facility. 3. The stability of the test or control article under the test conditions was not determined by the testing facility.

Result : Pilot: Mortality occurred within 4 hours following dosing: 5/5 males and 5/5 females at 2000 mg/kg, and 2/2 males at 1000 mg/kg. There were no

clinical observations noted.

Toxicity: Mortality occurred within 3 days of dosing: 5/5 males and 5/5 females at 400, 600, and 800 mg/kg, and 3/5 males and 5/5 females at 200 mg/kg. Clinical signs included: lethargy in males and females at all dose levels, piloerection in males and females, and crusty eyes in males at 200 mg/kg. The LD50/3 was calculated by pro bit analysis to be approximately 73.7 mg/kg for male and female mice.

Micronucleus Assay:

There was no treatment

- related mortality at any dose level. Clinical signs noted on the day of dosing included: lethargy in males and females at 60 mg/kg (highest dose). All other mice treated with test material and control

## 5. Toxicity Id 124

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61 / 89 articles appeared normal during the study. The number of micronucleated polychromatic erythrocytes per 2000 polychromatic erythrocytes in test

article

- treated groups was not statistically incr eased relative to their respective vehicle control in either male of female mice, regardless of dose

level or bone marrow collection time.

Test condition : ICR mice from Harlan Sprague

- Dawley, INC, were 6

- 8 weeks old at study initiation. For the pilot, t oxicity, and MNT assay, males ranged in weight from 27.4g to 31.9g, and females ranged from 23.0g to 28.0g. After arrival to the lab, the animals were monitored for parasites and infections, and were quarantined a minimum of 5 days. They were observed da ily for general health, food and water consumption patterns, and other conditions. The animals were deemed healthy prior to placement on study.

Mice were housed in an AALAC

- accredited facility with appropriate temperature, humidity, and photocycle for th e species. They were housed up to 5 per cage, and given food and water ad libitum. The feed and water contained no contaminants that influenced the study.

The mice were randomized and placed into assigned dose groups, and were identified by a uniquely n umbered ear tag.

Conclusion : Under the conditions of the assay described in this report, 2

- amino

- 2

- methyl

- 1

- propanol did not induce a sugnificant increase in the incidence of micronucleated polychromatic erythrocytes in bone marrow, and was

concluded to be negative in the micronucleus test using male and female

ICR mice. Reliability : (1) valid without restriction 1 (guideline, GLP study) Flag : Critical study for SIDS endpoint

01.06.2006 (43)

5.7 CARCINOGENICITY

5.8.1 TOXICITY TO FERTILITY

Type : other: OECD 421 Species : rat
Sex : male/female

Strain : other: CD

Route of admin. : oral feed Exposure period : 2 weeks before breeding through postpartum day 4 Frequency of treatm. : continuous Premating exposure period

Male : 2 weeks

Female : 2 weeks

Duration of test : Approximately 7 weeks No. of generation

studies :

Doses : 0, 100, 300, 1000 mg/kg/day Control group : yes, concurrent vehicle other: NOEL (repro effects) : = 100 mg/kg bw

other: NOEL (males, toxicity) : = 300 mg/kg bw

other: NOEL (females, toxicity) : < 100 mg/kg bw

Result : No effects on mating or conception. Liver effects in females at 100 mg/kg/day and higher, increased post

- implantation loss at 300 mg/kg/day and higher.

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62 / 89 Method : OECD Guide

- line 421

Year : 2005

GLP : yes Test substance : other TS: AMP

- HCl

Method : CD (Crl:CD(S D)IGSBR) rats were obtained from a commercial supplier and were approximately eight weeks of age at the time of study initiation. Each animal was evaluated by a laboratory veterinarian or a trained animal/toxicology technician, under the direct supervisio n of a lab veterinarian to determine their general health status and acceptability for study purposes upon arrival at the laboratory. The animals were housed 2

- 3 per cage in stainless steel cages, in rooms designed to maintain adequate conditions (tempera ture, humidity, and photocycle), and acclimated to the laboratory for approximately two weeks prior to the start of the study. They were offered a commercial diet and water ad libitum. During the study, animals were housed one per cage (prebreeding) or t wo per cage (one male and one female during breeding) in stainless steel cage in rooms designed to maintain adequate conditions (temperature, humidity, and photocycle). Dams were housed one per cage (with their litter) in plastic cages provided with corn cob nesting material from approximately day 19 of gestation and throughout the lactation phase of the study.

Animals were stratified by body weight and then randomly assigned to treatment groups using a computer program designed to increase the probabil ity of uniform group mean weights and standard deviations at the start of the study. Animals placed on study were uniquely identified via subcutaneously implanted transponders (BioMedic Data Systems, Seaford, Delaware) which were correlated to unique alph anumeric identification

numbers.

Groups of 12 male and 12 female CD rats were fed diets supplying 0 (control), 100, 300, or 1000 mg/kg/day of AMP. Males were exposed for at least two weeks prior to breeding and continuing throughout breeding for 37 day s. The females were exposed for two weeks prior to breeding, continuing through breeding (up to two weeks), gestation (three weeks), and lactation (four days). Effects on gonadal function, mating behavior, conception, development of the conceptus, partur ition, litter size, pup survival, sex, pup body weight and the presence of gross external morphological alterations were assessed. In addition, a gross necropsy and histopathology of the adults was conducted with an emphasis on organs of the reproductive system. Males were dosed via the diet for at least 14 days prior to mating, continuing throughout mating, for 37 days. Females were dosed by dietary exposure for 14 days prior to breeding, and continuing through breeding (up to two weeks), gestation (thr ee weeks), and lactation (four days).

Representative samples from the test diets were evaluated concurrently with the concentration verification analyses (see below) to ensure homogeneous distribution of the test material at the lowest and highest

concent rations in the feed at least once during the study. Preliminary stability analyses of the test material in rodent diets at concentrations of 0.0005% and 0.005% and 5.0% were initiated prior to the start of the

range

- finding study. Analysis of all test di ets from the first mix of the main study were initiated prior to the start of dosing using gas chromatography

mass spectrometry (GC

- MS) incorporating an internal standard to determine target concentrations.

Breeding Procedure Breeding of the adults comme nced after approximately two weeks of treatment. Each female was placed with a single male from the same dose level (1:1 mating) until pregnancy occurred or two weeks had elapsed.

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63 / 89 During the breeding period, daily vaginal lavage samples were evaluated

fo r the presence of sperm as an indication of mating. The day on which sperm were detected or a vaginal copulatory plug was observed in situ was considered day 0 of gestation. The sperm or plug

- positive (presumed pregnant) females then were separated from the male and returned to their home cages. If mating did not occur after two weeks, the animals were separated without further opportunity for mating.

Daily Observations A cage

- side examination was conducted twice daily, designed to detect significant cl inical abnormalities that were clearly visible upon a limited examination, and to monitor the general health of the animals for all males

pre

- exposure and weekly throughout the study. Clinical examinations were conducted on all females pre

- exposure and we ekly throughout the pre

breeding and breeding periods. Mated (sperm

- positive or plug

- positive) females received clinical examinations on GD 0, 7, 14 and 20. Females that delivered litters were subsequently evaluated on LD 0, 1 and 4, and on

additional da ys if warranted by observations made during daily cage

- side

examinations. Females that failed to mate or deliver a litter were examined weekly. Clinical observations included a careful, hand

- held examination of

the animal with an evaluation of abnormalit ies in the eyes, urine, feces, gastrointestinal tract, extremities, movement, posture, reproductive system, respiration, skin/hair

- coat, and mucous membranes, as well as an assessment of general behavior, injuries or palpable mass/swellings.

Body Weight s/Body Weight Gains Body weights for males were recorded on test days

- 1, 1, 4, 7, and weekly thereafter. Females were weighed on test days 1, 4, 7, and 14 during the

pre

- breeding period. During gestation, females were weighed on GD 0, 7,
14, and 20. Fe males that delivered litters were weighed on LD 1 and 4. Females that failed to mate or deliver a litter were not weighed during the gestation or lactation phases. Body weight gains were determined for the following intervals: GD 0

- 7, 7

- 14, 14

- 20, 0

- 20, and LD 1

- 4.

Feed Consumption Feed consumption for all animals was measured on test days 1, 4, 7 and 14 during the pre

- breeding period by weighing feed containers at the start and end of a measurement cycle. During breeding, feed consumption was

not meas ured in males or females due to co

- housing. Following breeding, feed consumption was measured weekly for males. For mated females, feed consumption was measured on GD 0, 7, 14, and 20. For females delivering litters, feed consumption was measured on LD 1 and 4. Feed consumption was not recorded for females that failed to mate or deliver a

litter.

Litter Data Females were observed for signs of parturition beginning on or about GD 20. In so far as possible, parturition was observed for signs of diffic ulty or unusual duration. The day of delivery was recorded as the first day the presence of the litter was noted and was designated as LD 0. Litters were examined as soon as possible after delivery. The following information

was recorded on each litter: the date of parturition, litter size on the day of parturition (day 0), the number of live and dead pups on LD 0, 1, and 4, and the sex and the weight of each pup on LD 1 and 4. Any visible physical abnormalities or demeanor changes in the neonates were recorded as they are observed during the lactation period. Any pups found dead or sacrificed in moribund condition were sexed and examined grossly, to the extent possible, for external and visceral defects and discarded.

Adult Necropsy A complete necrop sy of all the adults was performed. All males were necropsied on test day 38, while females that delivered litters were

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64 / 89 necropsied on LD 4. Females that did not deliver a litter were necropsied at least 24 days after the last day of the mating period. I n all cases, dosing continued until the day prior to sacrifice at which time the animals were fasted overnight. Fasted adult rats submitted alive for necropsy were anesthetized by the inhalation of carbon dioxide, weighed, and their tracheas exposed and c lamped. The animals were then euthanized by decapitation. A complete necropsy was conducted on all animals by a veterinary pathologist assisted by a team of trained individuals. The necropsy included an examination of the external tissues, and all orific es. The head was removed, the cranial cavity opened and the brain, pituitary and adjacent cervical tissues were examined. The eyes were examined in situ by application of a moistened microscope slide to each cornea. The nasal cavity was flushed via the nasopharyngeal duct and the lungs were distended to an approximately normal inspiratory volume with neutral, phosphate

- buffered 10% formalin using a hand

- held syringe and blunt needle. The skin was reflected from the carcass, the thoracic and

abdominal ca vities were opened and the viscera examined. All visceral tissues were dissected from the carcass, re

- examined and selected tissues

were incised. The uteri of all females were stained with a 10% solution of sodium sulfide stain for approximately two minu tes and were examined for the presence and number of implantation sites (Kopf et al., 1964). After evaluation, the uteri were gently rinsed with saline and preserved in neutral phosphate 10% formalin. Weights of the epididymides, kidneys, liver, and

test es were recorded, and organ:body weight ratios calculated.

Offspring Necropsy All pups surviving to LD 4 were euthanized by oral administration of sodium pentobarital solution, examined for gross external alterations, and then discarded. Any pups found dead were examined to the extent possible.

Histopathology Tissues with relevant gross lesions was conducted on all adult rats from the control and high

- dose groups. The histopathological examination of the testes included a qualitative assessment of sta ges of spermatogenesis. Examination of tissues from the remaining groups was limited to the liver (males and females), cervix, ovaries, oviducts, uterus, vagina, and relevant gross lesions. The histopathological examination of the testes included a quali tative assessment of stages of spermatogenesis. The presence and integrity of the 14 stages of spermatogenesis was qualitatively evaluated following the criteria and guidance of Russell et al. (1990). Microscopic evaluation included a qualitative assessm ent of the relationships between spermatogonia, spermatocytes, spermatids, and spermatozoa seen in cross sections of the seminiferous tubules. The progression of these cellular associations defined the cycle of spermatogenesis. In addition,

sections of b oth testes were examined for the presence of degenerative changes (e.g., vacuolation of the germinal epithelium, multinucleated giant cells, a decrease in the thickness of the germinal epithelium, a preponderance of Sertoli cells, sperm stasis, inflammator y changes, mineralization, and fibrosis). Selected histopathologic findings were graded to reflect the severity of specific lesions to evaluate: 1) the contribution of a specific lesion to the health status of an animal, 2) exacerbation of common natura lly occurring lesions as a result of the test material, and 3) dose

- response relationships for treatment related effects. Very slight and slight grades were used for conditions that were altered from the normal textbook appearance of an organ/tissue, but were of minimal severity and usually with less than 25% involvement of the parenchyma. This type of change would not be expected to significantly affect the function of the specific organ/tissue nor have a significant effect on the overall health of the a nimal. A moderate grade was used for conditions that were of sufficient severity and/or extent (up to 50% of the parenchyma) that the function of the organ/tissue may

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65 / 89 have been adversely affected, but not to the point of organ failure. The

health status of the animal may or may not have been affected, depending on the organ/tissue involved, but generally lesions graded as moderate would not be life threatening. A severe grade was used for conditions that were extensive enough to cause significant organ/t issue dysfunction or failure. This degree of change in a critical organ/tissue may be life threatening.

The above grading criteria were not sufficiently flexible to accurately characterize the liver microscopic vacuolization consistent with fatty change found in this study. Therefore, the following criteria were applied only to the observation of "liver vacuolization, consistent with fatty change, hepatocyte, multifocal": grade 1

- infrequently observed, grade 2

- occasionally observed, and grade 3

- rea dily observed.

STATISTICS AND CALCULATIONS Parental body weights and gestation and lactation body weight gains, litter mean body weights, feed consumption, and organ weights were first evaluated by Bartlett's test for equality of variances ( = 0.01; Wine r, 1971). Based upon the outcome of Bartlett's test, either a parametric or nonparametric analysis of variance (ANOVA) was performed. If the ANOVA was significant at alpha = 0.05, a Dunnett's test or the Wilcoxon

Rank

- Sum test with Bonferroni's correctio n (Miller, 1966) was performed (experiment

- wise alpha = 0.05). Statistical outliers were identified by a sequential test at alpha = 0.02 (Grubbs, 1969), and were excluded only for documented scientifically sound reasons. Feed consumption values were

excl uded from analysis if the feed had been spilled or scratched. Gestation length, average time to mating (precoital interval) and litter size were analyzed using a nonparametric ANOVA (Hollander and Wolfe, 1973). If the ANOVA was significant at alpha=0.05, the Wilcoxon Rank

Sum test (Hollander and Wolfe, 1973) with Bonferroni's correction was performed at experiment

- wise alpha = 0.05, two

- sided. The mating, conception, fertility and gestation indices (defined below) were analyzed by the Fisher exact probabi lity test (Siegel, 1956) with Bonferroni's correction at experiment

- wise alpha=0.05, two

- sided. Evaluation of the neonatal sex ratio on postnatal day 1 was performed by the binomial distribution test (Steel and Torrie, 1960) at alpha = 0.05, two

- sided. G ender was determined for pups born dead on postnatal day 0 and these data were

included in sex ratio calculations. Post

- implantation loss, pup survival indices and other incidence data among neonates were analyzed using the litter as the experimental unit by a censored Wilcoxon test (alpha = 0.05; Hollander and Wolfe, 1973) as modified by Haseman and Hoel (1974) with

Bonferroni's correction. Non

- pregnant females, females with resorptions only, or females found to be pregnant after staining of their uteri were excluded from the appropriate analyses. Both the Dunnett's test and Bonferroni's correction correct for multiple comparisons to the control to keep the experiment

- wise error rate at 0.05. Both were reported at the experiment

- wise alpha level. Becau se numerous measurements were statistically compared in the same group of animals, the overall false positive rate (Type I errors) was greater than the nominal alpha levels. Therefore, the final toxicologic interpretation of the data considered other

fact ors, such as dose

- response relationships, biological plausibility and consistency, and historical control values.

Calculation of Reproductive Indices Reproductive indices were calculated for female and male mating indices, male conception index, female a nd male fertility indices, gestation index, gestation survival index, post

- implantation loss, and days 1 and 4 pup

survival indices.

Result : All animals survived until scheduled termination. No treatment

- related

effects on behavior or demeanor were obse rved at any dose level during

the treatment

- period.

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66 / 89 There were no treatment related differences in the amount of feed consumed by any of the treated males or females when compared to their respective controls throughout the study. Gestation feed consumpti on of dams given 1000 mg/kg/day could not be compared due to a lack of viable

litters at this dose level.

No significant differences in body weights were observed for males at any

dose level tested. Pre

- mating body weights of treated females were not

dif ferent from controls. Dams given 300 mg/kg/day exhibited a non

statistically significant decrease (5.5%) in gestation day 20 body weight, and a statistically significant decrease in body weight gain between GD 14 and 20. This also led to a slight decreas e (not statistically identified) in body weight gain for the GD 0

- 20 interval. The mean GD 14

- 20 body weight gain in the 300 mg/kg/day group was 40% less than the control body weight gain during this period, which correlated closely with an approximate 50 % reduction in mean litter sizes observed in this group. Gestation body weights and body weight gains of dams given 100 mg/kg/day were not different from controls. Gestation body weights and body weight gains of females given 1000 mg/kg/day could not be compared due to a lack of viable litters at this dose level. There were no

treatment

- related differences in body weight or body weight gains during the lactation phase of the study.

There were no treatment

- related effects at any dose level on mating,

con ception, fertility, time to mating, gestation length or sex ratio. However, a marked increase in percent post

- implantation loss (embryo resorption) occurred in the 300 and 1000 mg/kg/day group females, with both the incidence, severity and likely timing o f this effect being dose

- related. In the 1000 mg/kg/day group, all twelve pregnant females showed evidence of complete litter resorption. In the 300 mg/kg/day group, four of twelve females exhibited complete litter loss, with the remaining eight females showing partial litter losses to varying degrees. Judging from the gross and microscopic appearance of the resorbed implantation sites, as well as the pattern of individual animal body weight gains during gestation, it is estimated that the embryonic deat hs occurred sometime toward the end of the second week of gestation (implantation is completed on GD 6; full term

is 21

- 22 days), with some of the high dose animals perhaps occurring slightly earlier. In the 100 mg/kg/day group, all reproductive parameter s including percent postimplantation loss were comparable to control values. One of 12 females in the 100 mg/kg/day group was initially considered non

pregnant based on the absence of implantation sites following sodium sulfide staining at necropsy. Howe ver, histological examination of the uterus revealed a very small amount of hemosiderin pigment in only one implantation site, suggesting that an embryo had implanted and then was resorbed almost immediately. This single case of very early resorption

doe s not fit with the timing of resorption seen in the 300 and 1000 mg/kg/day group, which occurred somewhat later. As pregnancies are not normally diagnosed histologically, there are no historical control data to directly address the significance of this ob servation. However, the likelihood that this was a spurious finding is supported by the fact that isolated occurrences of females that mate, but do no show evidence of embryo implantation following sodium sulfide staining are commonly encountered in repro ductive toxicity studies. Considering this, and the fact that all reproductive parameters in this group were unaffected by treatment, this single incident of litter resorption in the low dose group was considered spurious and unrelated to treatment

Observations recorded in the offspring occurred at low frequency and bore no relationship to treatment. There were no visible external morphologic alterations noted in any of the offspring delivered. There were no

treatment

- related effects on litter size in dams given 100 mg/kg/day. In the 300 mg/kg/day group, there were statistically identified decreases in mean

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67 / 89 number of pups born live, and mean number of pups on day 1 and 4 postpartum. These decreases in litter size were a function of the increase

in post

- implantation loss, as noted previously. No litters were produced by the 1000 mg/kg/day group females.

There were no treatment

- related effects on mean pup body weights in the 100 mg/kg/day group. Mean pup body weights in the 300 mg/kg/day group

w ere increased on days 1 and 4 postpartum, with the male mean pup weights identified as statistically different. These increases were most likely due to the decreased litter size at this dose level, as pup body weight varies inversely with litter size (Agn ish and Keller, 1997). Mean pup body weights of dams given 1000 mg/kg/day could not be evaluated due to a

lack of viable litters.

In the 1000 mg/kg/day males, mean body weight was increased approximately 5% when compared to the control group. Although not statistically identified, it was associated with statistically identified increases in absolute and relative liver and kidney weight. Similar body or organ weight effects were not observed in any of the female groups. However, it

should be noted that terminal body weight and organ weight data from the 1000 mg/kg/day group were excluded from analysis because they did not produce any viable litters and, therefore, could not legitimately be compared to the controls, which were in a post

- partum state. Al so excluded from analysis were four females in the 300 mg/kg/day group and one in the 100 mg/kg/day group which did not produce viable litters. The increased absolute and relative liver and kidney weights of the 1000 mg/kg/day males were statistically ide ntified and were outside of the historical control range for several studies, suggesting a treatment related

effect.

There were no treatment

- related gross pathologic observations. All

animals survived to the scheduled termination of the in

- life phase of the study.

Males given 1000 mg/kg/day and females given 100 mg/kg/day had

treatment

- related liver effects. Hepatocellular changes in 12/12 males of the 1000 mg/kg/day group were characterized by very slight, diffuse, cytoplasmic microvacuolization o f periportal hepatocytes. In general, the very slight effect was present in most of the periportal hepatocytes; however, in the three liver sections examined from different lobes (left lateral, middle, and right lateral), one was generally more noticeably affected. In general, the left and right lateral lobes were more affected than the middle lobe. The gradation of the effect was based upon the most severely affected lobe. The livers of a few male rats in the control, low and middle dose group were als o affected. It is likely the microscopic hepatocellular change in the 1000 mg/kg/day males was related to the significantly increased absolute and relative liver weight observed in this

group. There were 3/12 female control rats, and nearly all treated female rats with a similar microscopic hepatic effect. In addition to the aforementioned microscopic liver change in the various groups, another observation characterized by individual hepatocytes with vacuolated cytoplasm, suggestive of fatty change, was noted. The fatty change generally involved either the left or right lateral lobe most extensively. In no case was the middle lobe the most severely involved. In rats with a grade 2 or 3 involvement of hepatocytes with fatty change, the effect extended from the periportal region to the midzone of the lobule. Even in the most severely affected livers of either sex, the hepatocytes surrounding the central vein were microscopically normal. The fatty change in livers of 1000 mg/kg/day

males was observed at a higher frequency and severity when compared to lower dose groups and their respective control group. In the female rats of all treated groups, the incidence and severity of hepatic fatty change was

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68 / 89 increased compared to their respective controls.

In the female rats of all treated groups, the incidence and severity of hepatocellular fatty change was increased compared to their respective controls. The number of female rats in the 100 and 300 mg/kg/day groups was more frequently affected with a grade 2 and 3 hepatocellular fatty change in the 1000 mg/kg/day group. This may be due to the fact no litters were being nursed in the highest dose group. A clear dose response for the hepatic effect was not present in female 100 and 300 mg/kg/day groups. The preferential involvement of the left and right lateral lobes verses the middle lobe, in hepatocellular fatty change, may be due to normal variation in portal blood flow and/or normal variation in metabolic enzymes involved in lipid and triglyceride metabol ism. The microvacuolization and the fatty change in hepatocytes of even the most severely affected rats were not associated with evidence of increased hepatocellular necrosis or death. The observed microscopic hepatic effects were interpreted as reversib le. In the 1000 mg/kg/day male rats the microscopic liver effects observed were associated with increased absolute and relative liver weights. However, in the female rat livers of all dose levels were affected, despite

the absence of an increased liver we ight in this sex.

The microscopic hepatic fatty change was noted in essentially all 1000 mg/kg/day males and all treated groups of females; because a low
incidence was observed in controls of both sex, and low and middle dose males, it may represent nor mal variation associated with fasting. The microscopic vacuolization in the periportal hepatocytes seen in various groups of rats of both sexes may also be partially attributable to a variable degree of fasting prior to necropsy. The normal activity of c oprophagy in the rat could also contribute to periportal hepatocellular vacuolization. Therefore, greater amounts of glycogen would still remain in the periportal hepatocytes. Whether the effects within the livers of male and female rats can be totally a ttributed to indirect nutritional effects is uncertain. It is likely that the response in the liver was due to nutritional considerations and the test chemical. The presence of the test chemical in their feed likely accentuated this effect in high dose m ales, and all dose group of females, with subsequent fatty change also being observed.

An alternative explanation for the observed liver fatty change is related to the known potential of a number of aliphatic alcoholic amines to cause a deficiency in th e nutrient choline in rats and mice. AMP has been shown to disrupt phospholipid synthesis (Cosmetic Ingredient Review, 1990). As reviewed by Zeisel et al. (1995), choline requirements of pregnant rats in particular are substantially increased as the deve loping fetus is dependent upon maternal supplies of choline. The latter would explain the observed sensitivity of pregnant females to the liver fatty change relative to males.

AMP

- induced choline deficiency may also have accounted for the

increased incid ence of fetal resorbtions in the present study consistent with findings of resorptions in mice fed choline

- deficient diets.

There were no histopathological lesions per se in the uterus. However, in light of the aforementioned effect on post

- implantation loss, four sections of uterus were microscopically examined from each female rat to ascertain the presence of one or more implantation sites. In the control, low, and
middle dose animals which delivered a litter, the uterus contained small

numbers of indi vidual pigment laden macophages within the deep layer of the endometrial stroma and the myometrium, with a large aggregation of these pigmented cells in the myometrium associated with the mesometrial attachment. This is the normal appearance of a post

- par tum uterus. In the remaining middle and high

- dose females which had totally resorbed litters, the uteri did not have the typical microscopic findings of an implantation site usually located in the region of the myometrium associated with the

mesometrial a ttachment. However, a clearly recognizable change was

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69 / 89 present in the endometrium likely associated with a previous implantation. Immediately beneath the uterine mucosa, within the endometrial stroma, there were increased numbers of pigment laden macropha ges. In five high

- dose females, the pregnancies were detected by microscopic examination of the uterus. Overall, the microscopic uterine findings in rats that did not litter were consistent with embryonic death and subsequent resorption.

Normal estrou s cyclic activity in all the control and all treated female rats was observed microscopically in the tissues examined. Microscopic findings in the tissues examined from the adults did not reveal a likely mechanism for the observed effects on post

- implanta tion loss. However, the fatty changes in the liver of all dose levels of female rats and the 1000 mg/kg/day males does suggest a possible effect on normal lipid and triglyceride metabolism. Whether similar effects may have been present in

tissues of the rapidly developing fetus or involved in the normal formation of the placenta to maintain pregnancy was not determined in this study. Certainly the normal metabolism of lipids and triglycerides is a critical function for cellular membrane synthesis, and co uld have affected fetal viability. Endocrine glands were not microscopically examined in this study to determine whether a morphologic change was present.

Male reproductive organs and their accessory sex glands were unaffected by treatment. The microsco pic observations in tissues other than liver and uterus, were not interpreted to be due to treatment with the test chemical.

Conclusion : Dietary exposure of male rats to 1000 mg/kg/day of AMP caused increases in absolute and relative liver weights, accom panied by a very slight degree of microvacuolization of periportal hepatocytes, with or without vacuolization of hepatocytes consistent with fatty change. Females in all treatment groups exhibited similar histopathological changes in the liver,

but in the absence of an organ weight change. Absolute and relative kidney weights were increased in the 1000 mg/kg/day males, but these were not considered toxicologically significant due to the absence of histopathological changes. AMP had no effect on mating pe rformance or conception, but caused marked, dose

- related increases in post

implantation loss (embryo resorption). At the high dose level, all 12 pregnant females showed evidence of complete litter resorption (100%
post

- implantation loss), while at 300 mg/ kg/day, post

- implantation loss was 70% (vs. 10% in controls). Effects associated with, or secondary to the
post

- implantation loss increase at 300 mg/kg/day included decreased litter size, increased pup body weight, and decreased gestation body weight and body weight gain. There were no treatment related effects on reproductive performance in the 100 mg/kg/day group.

The no

- observed effect level (NOEL) for general toxicity in males was 300 mg/kg/day, while the general toxicity NOEL for females could not be determined, based upon the presence of very slight microscopic liver effects. The NOEL for reproductive effects was considered to be 100 mg/kg/day. Reliability : (1) valid without restriction 1 (GLP, guideline study) Flag : Critical study for SIDS e ndpoint

18.09.2006 (44)

5.8.2 DEVELOPMENTAL TOXICITY/TERATOGENICITY

Species : rat
Sex : female

Strain : other: CRL:CD(SD)

Route of admin. : dermal Exposure period : Gestation Day 6

- 20

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70 / 89 Frequency of treatm. : 6 hours per day

Duration of test : 1 5 days

Doses : 0, 30, 100, 300 mg/kg/day Control group : yes, concurrent vehicle

NOAEL maternal tox. : = 100

other: NOEL (developmental tox) : = 300

Method : OECD Guide

- line 414 "Teratogenicity"

Year : 2006

GLP : yes Test substance : as prescribed by 1.1

- 1.4

Method : Groups of 26 time

- mated female Crl:CD(SD) rats were administered AMP once daily by the dermal route at dose levels of 0, 30, 100, or 300 mg/kg/d ay for approximately

6 hours from GD 6

- 20. Dose solutions were prepared in deionized water at concentrations of 30, 100, and 300 mg/ml and pH adjusted to approximately 9.5 with HCl. The solutions were administered at a dose volume of 1 ml/kg body weigh t in order to achieve the targeted dose levels. Dose volumes were adjusted daily based on individual body weights. Dose solutions were prepared weekly during the course of the study. Homogeneity and stability were determined prior to study initiation, and concentration of the dose suspensions was

determined from the first mix.

Time

- mated female rats were obtained from a commercial supplier. Sexually mature adult rats were 10

- 11 weeks of age and weighed approximately 200

- 250 grams at study start.

Each animal was evaluated by the laboratory veterinarian to determine the general health status and acceptability for study purposes upon arrival at the laboratory. Rats were housed one per cage in stainless steel cages, in rooms designed to maintain adequate conditions (temperature, humidity, and photocycle) and acclimated for at least three days. Prior to dosing animals were acclimated to the wrapping material for approximately 2, 4, and 6 hours on GD 3, 4, and 5, respectively.

Animals were housed one per cage in stainless steel cages in rooms designed to maintain adequate environmental conditions for the species. Animals were provided feed and municipal water ad libitum. Rats were stratified by GD 0 body weight and then randomly assigned to treatment

gro ups using a computer program designed to increase the probability of uniform mean group weights and standard deviations at the start of the study. Animals that were placed on study were uniquely identified via subcutaneously implanted transponders (BioMed ic Data Systems, Seaford, Delaware) that were correlated to unique alphanumeric identification

numbers.

Sexually mature, adult virgin females were naturally mated with males of the same strain at the supplier's facility. Females were checked for in sit u copulation plugs the following morning and those found with such a plug were removed from the males' cages. The day on which a vaginal plug was detected was considered GD 0. GD 0 body weights were provided by the supplier, and maintained in the study r ecord. Rats arrived in the testing laboratory on GD 1 or 2.

A cage

- side examination was conducted twice daily, at approximately the same time each day to detect significant clinical abnormalities that were clearly visible upon a limited examination, and used to monitor the general

health of the animals. The animals were not hand

- held for these observations unless deemed necessary. In addition, all animals were observed for morbidity, mortality, and the availability of feed and water at least twice daily

.

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71 / 89 Following removal of the wrapping material at the end of each daily six

hour exposure period, the test sites were graded for erythema, edema, scaling, and fissuring.

Body weights were recorded on GD 0 by the supplier, and daily during the dosing per iod, and at necropsy (GD 21). Feed consumption were measured and statistically analyzed for the following intervals: GD 3

- 6, 6

- 9, 9

- 12, 12

- 15, 15

- 18, and 18

- 21.

On GD 21, all surviving females (not fasted) were euthanized by carbon

dioxide inhalation and a limited gross pathologic examination (necropsy) was performed. The sequence of the maternal necropsies was counterbalanced across groups (e.g., control, high, middle, low) to control for potential confounding influences of timing on fetal growth an d skeletal ossification. The maternal necropsy included an examination of the external tissues and all orifices. The stomach, liver, and kidneys were dissected from the carcass and were incised. Any obvious gross pathologic alterations were recorded, an d the weight of the liver, kidneys, and gravid uterus were recorded. The ratios of liver and kidney weights to terminal body weight were calculated. Representative sections of liver, kidneys, and gross lesions were preserved in neutral, phosphate

- buffere d 10% formalin. Microscopic examination of tissues was not conducted.

A detailed examination of the reproductive tract was performed and the number and position of implantations, viable fetuses, dead fetuses, and resorptions were recorded. Resorptions were classified as either "early" or "late" based on the presence (late resorption) or absence (early resorption) of grossly recognizable embryonic/fetal form, while a "dead fetus" would indicate a very recent death as evidenced by a lack external degener ative changes. For females with one or more viable fetuses, the number of ovarian corpora lutea were counted. The uteri of females lacking visible implantations were stained with a 10% aqueous solution of sodium sulfide (Kopf et al., 1964) and examined f or evidence of early resorptions in order to verify pregnancy status.

The sex of all fetuses was determined and the body weight of all viable fetuses recorded. All fetuses were given an external examination that included observations on body proportion s, the head and face (including closure of the palate), abdomen, spine, extremities, genitalia, rectum, and tail. At least one half of all the fetuses in each litter were chosen randomly via computer for visceral examination conducted by dissection under a low power stereomicroscope for evidence of visceral alterations. The visceral examination included observation of the thymus, trachea, esophagus, lungs, great vessels, heart (external and internal), liver, gastrointestinal tract, pancreas, spleen, kidne y (sectioned), adrenal glands, ureters, bladder, and reproductive organs. The heads of these fetuses were removed, placed in Bouin's fixative and serially sectioned to allow for inspection of the eyes, brain, nasal passages and tongue. The remaining

fetu ses not selected for visceral examination were then skinned, eviscerated, preserved in alcohol and double stained with Alcian Blue and Alizarin Red S for cartilage and bone. After staining, skeletons were macerated and cleared. A thorough evaluation of t he fetal skeleton was conducted on the remaining fetuses not selected for visceral examination.

All fetal alterations were classified as a variation or malformation. A variation is defined as a divergence beyond the normal range of structural

constitut ion that may not adversely affect survival or health. A malformation is defined as a permanent structural change that may adversely affect survival, development or function and/or which occurs at a relatively low incidence in the specific species/strain. The fetal examinations were conducted such that investigators were blind to treatment group assignment.

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72 / 89 Toxicokinetic Subgroup The first four presumed pregnant females from each dose group were selected for blood collection to evaluate systemic exposure following dermal administration of AMP. On the last day of dosing (GD20), following approximately three hours of dermal exposure, the designated animals were anesthetized using isoflurane and blood was collected from the orbital sinus. Approximately 0.5 ml of blood from each animal was collected into heparinized tubes. The samples were submitted to the analytical chemistry department for analysis and were stored frozen at approximately

- 80 C. AMP present in the blood was derivatized with pentafluorobenz oyl chloride
under basic conditions and extracted in toluene. Quantitation was performed utilizing an isotopically labeled internal standard (D6

- AMP) and matrix standards prepared in control blood. The limit of quantitation was 15 ng/g blood AMP.

STATI STICS AND CALCULATIONS Maternal body weights, maternal body weight gains, organ weights (absolute and relative), fetal body weights, and feed consumption were evaluated by Bartlett's test (alpha = 0.01; Winer, 1971) for equality of

variances. Based on the outcome of Bartlett's test, a parametric (Steel and Torrie, 1960) or nonparametric (Hollander and Wolfe, 1973) analysis of variance (ANOVA) was performed. If the ANOVA was significant at alpha = 0.05, analysis by Dunnett's test (alpha = 0.05; Winer, 1971 ) or the

Wilcoxon Rank

- Sum test (alpha = 0.05; Hollander and Wolfe, 1973) with Bonferroni's correction (Miller, 1966) were performed, respectively. Feed consumption values were excluded from analysis if the feed was spilled or

scratched. Frequency of pr e

- and post

- implantation loss (calculations shown below), and fetal alterations were analyzed using a censored Wilcoxon test (Haseman and Hoel, 1974) with Bonferroni's correction. The number of corpora lutea, implantations, and litter size were evaluated using a nonparametric ANOVA (alpha = 0.05) followed by the Wilcoxon Rank

- Sum test (alpha = 0.05) with Bonferroni's correction. Pregnancy rates were analyzed using the Fisher exact probability test (alpha = 0.05; Siegel, 1956) with Bonferroni's correction. Fetal sex ratios were analyzed using a binomial distribution test. Females lacking visible implantations at the scheduled necropsy were excluded from the appropriate analyses. Statistical outliers were identified, using a sequential method (alpha = 0.0 2; Grubbs, 1969), and if excluded, were excluded for sound scientific reasons. Both Dunnett's test and Bonferroni's correction correct for multiple comparisons to the control group to keep the experiment

- wise alpha at 0.05. Both were reported at the expe riment

- wise alpha level. Because numerous measurements were statistically compared in the same group of animals, the overall false positive rate (Type I errors) was greater than the nominal alpha levels. Therefore, the final interpretation of the data

wil l consider statistical analyses along with other factors, such as dose

response relationships and whether the results are consistent with other biological and pathological findings and historical control values.

Remark : Test material was utilized at a pH of approximately 9.5, reflecting the highest pH of most personal care and industrial formulations containing AMP. The test material represents an equilibrium between the HCl salt of AMP and the nonionized base ("AMP

- base"). As AMP

- base is expected to

be the primary form absorbed by skin, the test material in this study reflects the form of the material resulting in maximal exposure of humans from

dermal contact with an AMP

- containing product.

Result : In

- Life Observations Examinations performed on all a nimals during the course of the study

revealed no treatment

- related clinical findings. Dermal grading of the test site revealed a pattern of localized effects most pronounced in the 300 mg/kg/day dose group. The 300 mg/kg/day females showed significant

d ermal effects during the grading period, as compared to the other

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73 / 89 treatment groups. One 300 mg/kg/day female was found with very slight

edema from GD 9

- 13. This finding was not observed in any other dose group. Almost all of the 300 mg/kg/day females (9 2%) were observed with slight scaling, with nine animals (36%) progressing to moderate to severe scaling during the last half of the dosing period. One, two, and seven females at 0, 30, and 100 mg/kg/day, respectively, were also found with slight scaling, but this was not considered adverse, as the incidence was relatively low, as compared to the 300 mg/kg/day group and the finding resolved prior to necropsy in three of the seven 100 mg/kg/day animals. In addition, no animals at the 30 and 100 mg/kg/day g roups showed any signs of moderate to severe scaling. Scabbing was mainly observed at the 300 mg/kg/day dose level. Twenty 300 mg/kg/day females (77%) were noted with scabs at up to 25% of the test site. Over time, these the scabs progressed to include ever increasing areas of the test site, with one animal showing scabs covering most of the test site for the last week of dosing. The initial occurrence of scabbing varied considerably among the females, but there was a tendency for animals showing the gre atest amount of scabs to have developed these lesions earlier. With the exception of one control animal, no other females were observed with any scabbing. Overall, the dermal grading procedures revealed a treatment

- related effect at the high

- dose level of 300 mg/kg/day. The slight, transient scaling seen at lower dose levels was not considered to be toxicologically significant.

Body Weights/Body Weight Gains There were no statistically identified differences in the mean gestation body weight for female s at any dose level. Feed consumption was increased at 100 mg/kg/day during GD 3

- 9 interval. These statistically identified parameters were not considered treatment related, but attributed to relatively low control values.

There were no statistically i dentified differences in mean terminal body weight, mean liver and kidney weights, or relative liver and kidney to body weight ratios in the treatment groups, as compared to the control. Gross pathological findings were spurious in nature and not attribut ed to

treatment.

There were no significant treatment related effects on pregnancy rate, litter size, numbers of corpora lutea or implantations, percent preimplantation loss, fetal sex ratios, fetal body weights, or gravid uterine weights at any

dose level . Mean percent postimplantation loss was statistically decreased in the 300 mg/kg/day group, but this was not considered to be test article

related, as the value was lower than seen in the control group. One female in the 30 mg/kg/day group was found to be non

- pregnant, as confirmed by ammonium sulfide staining.

There were no statistically significant differences in the incidence of any fetal alteration (malformation or variation) in any of the treated groups, as compared to the control. There were no e xternal fetal alternations observed. Any reported findings were sporadic in nature, and were not considered toxicologically relevant or treatment related.

Toxicokinetic Subgroup Blood samples were obtained from four presumed pregnant females on the

las t day of dosing following approximately three hours of dermal application. The results indicate dermal absorption of AMP in a dose

responsive manner. The disproportionately higher mean blood concentration of AMP at 300 mg/kg/day may have been the result of a compromised skin barrier, as evidenced by significant dermal effects (scabbing and scaling) at the test site for that group. At this dose level, one female (animal # 7169) was noted with a blood value approximately three

times that of the others. Th e increase in the blood value for this animal is

## 5. Toxicity Id 124

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Date 30.11.2006

74 / 89 likely the result of a greater degree of compromise to the skin barrier, as evidenced by a greater degree of scabbing (up to 50%) at the test site on the day of blood collection, as compared to the other ani mals in the group.

Conclusion : Dermal administration of 300 mg/kg/day of AMP produced significant effects at the test site, as evidenced by scabbing (77% affected) and moderate to severe scaling (35% affected). The dermal finding of slight scaling at 30 and 100 mg/kg/day was not considered adverse, as the observation was transient in nature and relatively low in incidence. There was no evidence of test article related systemic maternal or developmental toxicity at any dose level tested. Under the condi tions of this study, the NOAEL for maternal toxicity based on dermal effects was 100 mg/kg/day. The NOEL for developmental toxicity was 300 mg/kg/day, the highest dose level tested. Analyses of blood samples confirmed systemic exposure to

AMP in a dose

- r esponsive manner, although the study was not designed to quantify percent absorption. Reliability : (1) valid without restriction 1 (GLP, guideline study) Flag : Critical study for SIDS endpoint

07.09.2006 (45)

Species : rat
Sex : female

Strai n : Sprague

- Dawley

Route of admin. : dermal Exposure period : GD 6

- 20 Frequency of treatm. : daily

Duration of test : 6 hours per day

Doses : 0, 100, 300 or 400 mg/kg/day Control group : yes, concurrent vehicle

Method : Year : 2005

GLP : yes Tes t substance : as prescribed by 1.1

- 1.4

Method : The purpose of this study was to make a preliminary evaluation of the maternal toxicity and embryo/fetal lethality potential of 2

- methyl

- 2

- aminopropanol hydrochloride (AMP

- HCl) in Crl:CD(SD) rats followin g repeated dermal administration. Groups of eight time

- mated female rats

received AMP

- HCl by occluded dermal application, six hours/day at targeted dose levels of 0, 100, 300 or 400 mg/kg/day on gestation days (GD) 6

- 20. In

- life parameters evaluated for all groups included clinical observations, body weight, body weight gain, feed consumption, and dermal grading of the test site. Blood samples were collected approximately three hours after dermal application, from 2

- 3 rats/group on

GD 20 to evaluate derm al absorption of AMP

- HCl. On GD 21, all surviving rats were euthanized and examined for gross pathologic alterations. Liver and kidney weights, the number of corpora lutea, implantations, resorptions, and live/dead fetuses were recorded.

Result : Dermal administration of AMP

- HCl at dose levels of 100, 300 and 400 mg/kg/day resulted in no maternal toxicity and no indication of adverse embryo/fetal effects. Administration of the test material by the dermal route was well tolerated by all animals, without s ignificant adverse dermal effects. Analyses of blood samples taken on the last day of dosing

indicated a dose

- responsive increase in systemic exposure, although the study was not designed to quantify percent absorption. Based on these
results, the no

- obs erved

- effect level (NOEL) for maternal and embryo/fetal development was 400 mg/kg/day, which was the highest dose level tested. As no significant adverse dermal effects were noted in the developmental toxicity probe study, a two

- week dermal follow

- up stu dy using male and

non

- pregnant female rats was initiated (Phase II). Solubility issues were

encountered with AMP

- HCl when it was prepared again using the

## 5. Toxicity Id 124

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Date 30.11.2006

75 / 89 hydrochloride salt. As a result, the base form of AMP was used in the prepration of the dose formula tions for this study, and this alleviated the difficulties with solubility. AMP base (pH 9.5) at 300, 400 and 500 mg/kg/day was administered once daily, by the dermal route to the animals. Significant adverse dermal effects (scabbing, redness,

tenderness to touch) were noted at the two highest dose levels (400 and 500 mg/kg/day) after a few days of dosing. This resulted in the immediate termination of treatment to these two dose groups. However, the low

- dose of 300 mg/kg/day showed only slight dermal ef fects after 11 days of dosing and was, therefore, used as the high

- dose level for the definitive developmental toxicity study.

Based on the results obtained from this follow

- up work and the developmental toxicity probe study, dose levels of 0, 30, 100, or 300 mg/kg/day of AMP base were selected for a definitive dermal developmental toxicity study in rats. Reliability : (1) valid without restriction 1 (GLP study of acceptable quality with documentation sufficient for assessment)

07.09.2006 (46)

5.8.3 TOXICITY TO REPRODUCTION, OTHER STUDIES

Type : other: dietary toxicity

In vitro/in vivo : In vivo Species : dog

Sex : male/female

Strain : Beagle

Route of admin. : oral feed Exposure period : 1 year Frequency of treatm. : continuous

Duration of test : Doses : 0, 1.1, 11.0, 110.0 ppm Control group : yes, concurrent vehicle

Result : No effect on reproductive organs

Method : Year : 1990

GLP : Test substance : as prescribed by 1.1

- 1.4

Result : There were no general in

- life observations made for any treated or control animal. There is no statistically significant differences in body weights for any of the animals throughout the study.

There were no effects on organ weights or organ/body weight ratios that

were attributed to test mater ial administration by the authors. Reproductive organs that were examined included: testes, ovaries, and uteri.

At necropsy, evaluation of the tissues revealed isolated observations, none

of which were attributed to the administration of the test materia l. There were no neoplastic observations in any of the dogs sacrificed at either 6 months or one year of administration in their diets. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and

ac ceptable for assessment)

01.06.2006 (33)

Type : other: dietary toxicity

In vitro/in vivo : In vivo Species : dog

Sex : male/female

## 5. Toxicity Id 124

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- 5

Date 30.11.2006

76 / 89 Strain : Beagle

Route of admin. : oral feed Exposure period : 90 days Frequency of treatm. : continuous

Duratio n of test : 90 days

Doses : Control group : Method : Year : 1981

GLP : Test substance : as prescribed by 1.1

- 1.4

Result : Physical Observations Included emesis, loose stool, alopecia, and slight penile or vaginal discharge. These findings oc curred sporadically in the control and/or treated animals and are not considered by the authors to be treatment

related.

Urinalysis There is no evidence of a compound

- related effect on any of the urinalysis parameters studied.

Reproductive organs evaluat ed: Testes (without epididimus), ovaries, and

uterus.

Organ Weight/ Organ:Body Weight Ratios Organ weight and organ/body weight differences were marginal and appear unrelated to the administration of the test material.

Pathology

There were no observation s noted in any reproductive organ upon gross and microscopic evaluation. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment)

01.06.2006 (39)

Type : other: die tary toxicity

In vitro/in vivo : In vivo Species : rat
Sex : male/female

Strain : other: CD

Route of admin. : oral feed Exposure period : 8 weeks Frequency of treatm. : continuous

Duration of test : 8 weeks

Doses : 1000, 2000, 4000, 8000, 16000 pp m Control group : yes, concurrent vehicle

Method : Year : 1976

GLP : Test substance : as prescribed by 1.1

- 1.4

Result : Rats in the 16000ppm group gained slightly less weight as compared to control rats. Gains in the other dose groups were simi lar to the control. Food consumption patterns were similar for all animals.

At necropsy, all 16000ppm surviving rats, and all rats at the 8000ppm dose level were necropsied. Two males and two females from each of the other groups were also examined. Alopecia and focal skin erosions and ulcerations found in rats at the 16000ppm dose level were viewed as possibly compound related. Hepatocyte vacuolation descrived in rats from

## 5. Toxicity Id 124

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Date 30.11.2006

77 / 89 all the treated groups, with increasing severity at the higher dose levels,

w as considered compound related. Microscopic skin lesions found at the 16000ppm dose level, like the corresponding gross lesions, were viewed as possibly compound related.

There were no gross or microscopic observations noted in epididimus,
uteri, ovaries , or testes. Reliability : (2) valid with restrictions

2e

01.06.2006 (34)

5.9 SPECIFIC INVESTIGATIONS

Endpoint : Mechanistic Studies Study descr. in chapter : Reference : Type : other: In vitro using Chinese hamster ovary cells Species : Sex : Strain : Route of admin. : No. of animals : Vehicle : Exposure period : Frequency of treatm. : Doses : 0.8, 4, 8, 16, 24, and 32 mM (100, 500, 1000, 2000, 3000, and 4000 mg/L, respectively) in the first Trial and 0.8, 4, 8, 16, 32, 4 8, and 80 mM (100, 500, 1000, 2000, 4000, 6000, and 10000 mg/L, respectively) in the second

trial Control group : Observation period : Result : Method : Year : 2006

GLP : Test substance : as prescribed by 1.1

- 1.4

Method : The potential of A MP·HCl to inhibit the uptake of the essential nutrient choline in cultured mammalian cells was determined. Effects of AMP upon choline uptake by CHO cells were evaluated in actively growing cultures exposed to 3H

- choline and a range of concentrations of A MP·HCl (0

- 32 mM; 0

- 4000 mg/L) for approximately 10 minutes. Concentrations of AMP·HCl used were based upon the results of a cytotoxicity assay and encompassed concentrations ranging from nontoxic to moderately toxic following 48 hours exposure. The pH o f all dosing solutions was adjusted to that of the culture medium. Following treatment, cells were harvested, washed and lysed. The protein concentration of the lysate and 3H content were determined. Choline uptake as 3H activity was calculated on a per mg protein basis and treated and control data were compared statistically. In addition, the approximate effective concentration eliciting a 50% inhibition of uptake (EC50) was calculated and compared with that of several other alcohol amines.

Descriptiv e statistics (means and standard deviations) for choline uptake were reported as total 3H counts per mg protein. Control and treated values for choline uptake were evaluated by Bartlett's test (alpha=0.01; Winer, 1971) for equality of variances. Explorat ory data analysis was performed by a parametric analysis of variance (ANOVA) (alpha=0.05; Steel and Torrie, 1960) and followed, if significant, by Dunnett's test (alpha=0.05; Winer, 1971). Dunnett's test corrects for multiple

## 5. Toxicity Id 124

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Date 30.11.2006

78 / 89 comparisons to the control an d the experiment

- wise alpha level was reported. Statistical outliers were identified by a sequential test (alpha=0.02; Grubbs, 1969).

Result : A dose

- response in cell viability versus concentration of AMP·HCl was observed in both toxicity trials. Surviv al relative to untreated controls ranged from 100+% to 61

- 68% over a concentration range of 0.04

- 32 mM. A 29% and 5.5% survival was noted at 48 and 80 mM, respectively, in trial II. The positive control consisting of 5% ethanol resulted in a survival of only 28

- 36% of untreated controls in both trials. No effects upon pH or osmolarity of the treated cultures were observed. Based upon these findings, a concentration range of 0.4 to 32 mM AMP·HCl was evaluated for effects upon 3H

- choline uptake by CHO cel ls.

Uptake of 3H

- Choline There were no significant differences in protein content between treated and control cultures indicating no significant loss of cells in the 10

- minute dosing period, including the high dose which resulted in cell numbers 61

- 68% of controls following 48 hours treatment. AMP·HCl was found to

cause a dose

- related decrease in 3H

- choline uptake over the 40 to 80 fold concentration ranges examined; 0.8

- 32 mM in Choline Trial I and 0.4

- 32 mM in Choline Trial II. A statistically signific ant decrease in the uptake of

3H

- choline was achieved at concentrations of 4 mM (65

- 81% of control) to 32 mM (30

- 36% of control) in both Trials. AMP·HCl appears to be a relative weak inhibitor of choline uptake in vitro with an approximate EC50

of 8.71

- 1 4.79 mM. It is approximately 10

- 17 fold less active than triethanolamine (EC50 ~0.89 mM), approximately 17

- 28 fold less active than diisopropanolamine (EC50 ~0.52 mM), and approximately 44

- 74 fold less active than diethanolamine (EC50 ~0.20 mM) in the sam e CHO cell based in vitro assay.

Conclusion : It was concluded that AMP·HCl has the potential to inhibit uptake of the essential nutrient choline in cultured CHO cells in the absence of significant toxicity. This activity, however, is relatively weak whe n compared to several other alcohol amines under similar assay conditions. Reliability : (1) valid without restriction 1 (meets generally accepted scientific standards and is described in sufficient detail)

02.06.2006 (47)

Endpoint : Mechanistic Studies Study descr. in chapter : Reference : Type : other: in vivo Species : rat
Sex : female

Strain : Sprague

- Dawley

Route of admin. : dietary

No. of animals : Method : Year : 2006

GLP : yes Test substance : as prescribed by 1.1

- 1.4

M ethod : The potential of dietary HCl salt of 2

- amino

- 2

- methyl

- 1

- propanol (AMP

- HCl) to depress hepatocellular levels of the essential nutrient choline in pregnant rats was evaluated. Two groups of 6 female Crl:CD(SD) rats each were fed diets containing 0 ( control) or 300 mg/kg/day of AMP

- HCl for approximately two weeks prior to breeding, continuing throughout breeding (up to two weeks), and two weeks into gestation (GD 0

- 13). Clinical observation, body weight, and feed consumption data were collected. On GD 14, the females were euthanized and samples of liver were excised and frozen in liquid nitrogen or saved in 10% formalin. Uteri were examined and the number of live implants and resorptions were

## 5. Toxicity Id 124

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Date 30.11.2006

79 / 89 determined. Formalin fixed liver underwent histopatholog ical examination following standard hematoxylin and eosin staining and osmium tetroxide treatment for identification of lipid. Frozen liver was analyzed for levels of choline (Cho), betaine, glycerophosphocholine (GPCho), phosphocholine (PCho), phosphatid yl choline (PtdCho), and sphingomyelin (SM).

Result : Dietary administration of 300 mg/kg/day of AMP

- HCl resulted in no adverse clinical observations, mean body weight, or feed consumption effects.
However, AMP

- HCl ingestion caused an increased postimpla ntation loss (67.0% in treated versus 7.4% in control rats), and a slight increase in hepatic lipid vacuolation relative to controls. These changes to varying degrees, correlated with an approximately 20

- 25% statistically identified decrease in hepatic le vels of PCho which is the primary storage pool of choline. Levels of GPCho, indicative of conversion of PtdCho to Cho, were

elevated 2

- 3 fold relative to control dams. Hepatic levels of remaining metabolites were either unchanged or minimally changed rel ative to

controls.

Conclusion : The results of this study, when combined with the known choline depression caused by pregnancy, suggest that previous findings of lipid accumulation and elevated resorptions in an OECD

- 421 study of AMP

- HCl in rats (Carney e t al., 2005) may be a consequence of, and secondary to, PCho deficiency, respectively. Reliability : (1) valid without restriction 1 (GLP study, based in part on standard test methods, well

- documented and acceptable for assessment)

16.10.2006 (48)

5 .10 EXPOSURE EXPERIENCE

Type of experience : Human

Method : Skin prick tests and patch tests with the test material and common inhalant allergens were performed on all subjects. AMP

- 100 was dissolved in both water and ethyl alcohol separately at 0. 1%, 0.5%, 1%, 2%, 5%, 10%, 20%. Patch tests were applied on the back and read at 48 and 72 hours. Positive results were recorded for erythema, oedema, and blisters.

Result : The prick and patch tests were all negative except for one positive reaction

to Dermatophagoides pteronyssinus in a worker on a different production line. All patch tests carried out with substances used in the production line proved negative except for AMP

- 100 at 10% and 20% dilutions in water and ethyl alcohol. All positive resul ts manifested as erythema with oedema without blistering.

Positive results at 10% and 20% may be considered as irritative responses. There was lack of response to the lower doses (that would have been considered allergenic responses) noted in both the asymptomatic and symptomatic subjects.

Test condition : Two cases of airborne contact dermititis were noted in a cosmetic company during the hair dye production using AMP

- 100. Subjects suffered for months from periorbital oedema and facial erythema which worsened during work hours and improved with time away from work. Eight additional asymptomatic workers from the same company were studied concurrently.

Conclusion : The authors conclude that under these conditions, AMP

- 100 causes

airborne irritative co ntact dermatitis, favored in this study by elevated production temperatures and high ambient humidity. Reliability : (2) valid with restrictions 2 (meets generally accepted scientific standards, well

- documented, and acceptable for assessment)

01.06.20 06 (49)

## 5. Toxicity Id 124

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Date 30.11.2006

80 / 89 5.11 ADDITIONAL REMARKS

## 6. Analyt. Meth. for Detection and Identification Id 124

- 68

- 5

Date 30.1 1.2006

81 / 89 6.1 ANALYTICAL METHODS

6.2 DETECTION AND IDENTIFICATION

## 7. Eff. Against Target Org. and Intended Uses Id 124

- 68

- 5

Date 30.11.2006

82 / 89 7.1 FUNCTION

7.2 EFFECTS ON ORGANISMS TO BE CONTROLLED

7.3 ORGANISMS TO BE PROTECTED

7.4 USER

7.5 RESISTANCE

## 8. Meas. Nec. to Prot. Man, Animals, Environment Id 124

- 68

- 5

Date 30.11.2006

83 / 89 8.1 METHODS HANDLING AND STORING

8 .2 FIRE GUIDANCE

8.3 EMERGENCY MEASURES

8.4 POSSIB. OF RENDERING SUBST. HARMLESS

8.5 WASTE MANAGEMENT

8.6 SIDE

- EFFECTS DETECTION

8.7 SUBSTANCE REGISTERED AS DANGEROUS FOR GROUND WATER

8.8 REACTIVITY TOWARDS CONTAINER MATERIAL

## 9. References Id 124

- 68

- 5

Date 30.11.2006

84 / 89 (1) The Merck Index (Eleventh Edition). Merck & Co., INC. Rahway, NJ, USA. 1989.

(2) Budavari, S. (1989) The Merck Index (Eleventh Edition).

Merck & Co., INC. Rahway, NJ, USA. Not GLP, Published

(3) Wilson, L., Morrison, R., and Olson, J. (2005) The Vapor Press ure of 2

- amino

- 2

- methyl

- 1

-

propanol (AMP) at 20 Deg.C. The Dow Chemical Company Report No: SC AL SC

- 2005

- 000140 Not GLP / Unpublished

(4) Howard, P.H. and Meylan, W.M. (1997) Handbook of Physical Proporties of Organic

Chemicals.

Environmental Sciences Center, Syracuse Research Corp. Lewis Publishers, New York, NY, USA.

Not GLP / Published

(5) Lhotak, H. (1996) Determination of Partition Coefficient Pow (Octanol/Water) According to

OECD Method 107 of Six Angus Products. The Dow Chemical Company Repo rt No: DR

- 0366

- 3498

- 013 Not GLP / Unpublished

(6) Howard, P.H. and Meylan, W.M. (1997) Handbook of Physical Proporties of Organic

Chemicals. Environmental Sciences Center, Syracuse Research Corp. Lewis Publishers, New York, NY, USA.

Not GLP / Publishe d

(7) Lewis, G.E. (1993) Determination of Basicity of 2

- Amino

- 2

- methyl

- 1

- propanol (AMP+),

Ethyl

- 1,3

- Propanediol (AEPD+), 2

- Amino

- 1

- butanol, Tris(Hydroxymethyl)Animomethane,

and 3

- Amino

- 1,2

- Propanediol. The Dow Chemical Company Report No: 933420

Not G LP / Unpublished

(8) Littel et.al. (1990) Dissociation Constants of Some Alkanolamines at 293, 303, 318, and

333K. J. Chem. Eng. Data 35:276

- 277.

Not GLP / Published

(9) Gonsior, S. (2006) Prediction of the Environmental Distribution and Transport Between

Environmental Compartments for 2

- amino

- 2

- methyl

- 1

- propanol Using Level I and Level III Fugacity Models. Report DR

- 0001

- 9476

- 004 of the Dow Chemical Company. Not GLP / Unpublished

(10) Harris, G.W. and Pitts, J.N. (1983) Rates of Reaction of H ydroxyl Radicals with 2

-

(Dimethylamino)ethanol and 2

- Amino

- 2

- methyl

- 1

- propanol in the Gas Phase at 300+

- 2K.

Environ. Sci. Technol. 17:50

- 51.

Not GLP / Published

(11) Larson, R.A. and Weber, E.J. (1994) Reaction Mechanisms in Environmental Organic

Chemist ry. Lewis Publishers, CRC Press, INC., Boca Raton, FL, USA.

Not GLP / Published

(12) Freitag, D. and Korte, F. (1989) Investigations of the Ecotoxicological Behavior of AMP

90, Tris Nitro, Amine CS

- 1246 and Alkaterge T. Technical University Munich, D epartment of Ecological Chemistry. The Dow Chemical Company Report No: (not assigned) Not GLP / Unpublished

## 9. References Id 124

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85 / 89 (13) Parekh, C. (1980) Acute Toxicity Effects of AMP

- 95 on Bluegill Sunfish, Ninety

- six Hour

LC50. Analytical Bio Chemistry Laboratories

The Dow Chemical Company Report No: DR

- 0309

- 4391

- 002 GLP / Unpublished

(14) Young, B.E. and Tapp, J.F. (1983). The Toxicity of AMP 95 to the brown shrimp (Crangon

crangon) and plaice (Pleuronectes platessa). Imperial Chemical Industries, PLC.

The Dow Chemica l Company Report No: DR

- 0309

- 4391

- 001 Not GLP / Unpublished

(15) Parekh, C. (1980) AMP

- 95 Acute Toxicity Effects on Daphnia, 48 Hr LC50.

Analytical Bio Chemistry Laboratories The Dow Chemical Company Report No: DR

- 0309

- 4391

- 003 GLP / Unpublished

(16) Young, B.E. and Tapp, J.F. (1983). The Toxicity of AMP 95 to the brown shrimp (Crangon

crangon) and plaice (Pleuronectes platessa). Imperial Chemical Industries, PLC. The Dow Chemical Company Report No: DR

- 0309

- 4391

- 001 Not GLP / Unpublished

(17) Sch afer, et.al. (1983) The Acute Oral Toxicity, Repellency, and Hazard Potential of 998

Chemicals to One of More Species of Wild and Domestic Birds. Arch. Environ,. Contam. Toxicol. 12:355

- 382.

Not GLP / Published

(18) Bridges, R.G. and Ricketts, J. (196 5). The Incorporation of 1

- aminobutanols into the

Phospholipids of Housefly (Musca domestica) Larvae. J. Insect Physiol. 11: 225.

Not GLP / Published

(19) Yue, K.T., Mulford, D.J., and Russell, P.J. (1970). The Metabolism of 2

- Amino

- 2

-

methylpropanol in Young Rats. The Department of Biochemistry, University of Kansas Medical Center, Kansas City, Kansas, USA. The Archives of Biochemistry and Biophysics (136:47

- 53).

Not GLP / Published

(20) Machle et.al. (1940) The Physiological Response of Animal s to Some Simple

Mononitroparaffins and to Certain Derivatives of These Compounds. Journal of Industrial Hygiene and Toxicology V22, No8 (315

- 332)

Not GLP / Published

(21) Hill Top Research (1955) Acute Toxicities of Amines.

Hill Top Research Co.

The D ow Chemical Company Report No: DR

- 0309

- 4391

- 015 Not GLP / Unpublished

(22) Commercial Solvents Corporation (1975) Bioassay Report, Pharmacology Laboratories

Research and Development Number 8945. Commercial Solvents Corporation The Dow Chemical Company Re port No: DR

- 0001

- 9476

- 006 Not GLP, Unpublished

(23) IMC (1976) P

- 1826

International Minerals Corporation The Dow Chemical Company Report No: DR

- 0001

- 9476

- 005 Not GLP, Unpublished

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86 / 89 (24) Power, C. (1976) Acute Oral Toxicity of P

- 1826 (AMP) in the Rat.

Pharmacology Laboratories, Animal Science Research The Dow Chemical Company Report No: DR

- 0309

- 4391

- 014 Not GLP / Unpublished

(25) Parekh, C. (1980) Dermal Toxicity of P

- 1826 (AMP).

Pharmacology Laboratories Animal Science Research

The Dow Chemical Com pany Report No: DR

- 0309

- 4391

- 011 GLP / Unpublished

(26) Kaltenbach, J.P., Ganote, C.E., and Carone, F.A. (1979) Renal Tubular Necrosis Induced

by Compounds Structurally Related to D

- Sterine. Experimental and Molecular Pathology, 30(2):209

- 214.

Not GLP / Published

(27) Power, C. (1975). AMP Regular and AMP

- 95

- Eye Irritation Tests

- Rabbits.

Commercial Solvents Corporation The Dow Chemical Company Report No: DR

- 0.09

- 4391

- 013 Not GLP / Unpublished

(28) Sausaman, D. (1975) Skin and Eye Irritation D ata for AMP, AMP

- 95, and AEPD.

The Dow Chemical Company Report No: DR

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- 4391

- 012 Not GLP / Unpublished

(29) Parekh, C. (1982) Topical Sensitization Potential of AMP (P

- 1826).

Bio/dynamics, Inc. The Dow Chemical Company Report No: DR

- 0309

- 4391

- 010

N ot GLP / Unpublished

(30) Parekh, C. (1982) Intradermal Sensitization Potential of AMP (P

- 1826).

Bio/dynamics, Inc. The Dow Chemical Company Report No: DR

- 0309

- 4391

- 009 Not GLP / Unpublished

(31) Pittz, E. (1977) A 90

- Day Safety Evaluation Study of 2

- Amino

- 2

- methyl

- 1

- propanol (AMP)

in Rats. The Institute of Comparative and Human Toxicology, Albany Medical College, Albany, NY. The Dow Chemical Company Report No: DR

- 0309

- 4391

- 023 Not GLP / Unpublished

(32) Parekh, C. (1981) A Three Month In Diet T oxicity Study of 2

- Amino

- 2

- methyl

- 1

- propanol in

Dogs. Bio/dynamics, Inc. The Dow Chemical Company Report No: DR

- 0309

- 4391

- 017 Not GLP / Unpublished

(33) Griffin, T. (1990) A One

- Year Oral Toxicity Study of AMP in Dogs (Amended by the original

author o n April 20, 1993). Coulston Research Incorporated, Alamogordo, NM. The Dow Chemical Company Report No: DR

- 0309

- 4391

- 004 Not GLP (contains an audit certificate) / Unpublished

(34) Goldenthal, E. (1976) Eight Week Tolerance Study in Rats.

(conducting l ab>>) The Dow Chemical Company Report No: DR

- 0309

- 4391

- 022 Not GLP / Unpublished

(35) Goldenthal, E. (1976) Eight Week Tolerance Study in Mice. (conducting lab>>)

The Dow Chemical Company Report No: DR

- 0309

- 4391

- 018 Not GLP / Unpublished

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87 / 89 (36) Goldent hal, E. (1976) Twenty Eight Day Dietary Range Finding Study in Dogs.

(conducting lab>>) The Dow Chemical Company Report No: DR

- 0309

- 4391

- 020 Not GLP / Unpublished

(37) Pittz, E. (1977) A Study on the Toxic Effects of 2

- amino

- 2

- methyl

- 1

- propanol in Ra ts and

Monkeys. A report of the Institute of Comparative and Human Toxicology, Center of Experimental Pathology and Toxicology, Albany Medical College, Albany, NY.

(38) Detwiler

- Okabayashi, K. and Shaper, M. (1996) Respiratory Effects of a Synthetic

Met alworking Fluid and its Components. Arch. Toxicol. (1996) 70:195

- 201.

Not GLP / Published

(39) Parekh, C. (1981) A Three Month In Diet Toxicity Study of 2

- Amino

- 2

- methyl

- 1

- propanol in

Rats. Bio/dynamics Inc, East Millstone, NJ.

The Dow Chemical Re port No: DR

- 0309

- 4391

- 021 Not GLP / Unpublished

(40) Wagner, V. (1996) Salmonella/Escherichia Coli Plate Incorporation Mutagenicity Assay

with a Confirmatory Assay. Microbiological Associates, INC. The Dow Chemical Company Report No: DR

- 0309

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- 008

GL P / Unpublished

(41) San, R. and Clark, J. (1997) In Vitro Mammalian Cell Gene Mutation Test with an

Independent Repeat Assay. Microbiological Associates, INC. The Dow Chemical Company Report No: DR

- 0309

- 4391

- 005 GLP / Unpublished

(42) Baldwin, R. (1 976) Mutagenicity Evaluation of P

- 1826 (AMP).

Litton Bionetics, INC, The Dow Chemical Company Report No: DR

- 0309

- 4391

- 007 Not GLP / Unpublished

(43) Gudi, R. (1998) Mammalian Erythrocyte Micronucleus Test (2

- amino

- 2

- methyl

- 1

- propanol).

MA Bioservic es, INC. The Dow Chemical Company Report No: DR

- 0309

- 4391

- 006 GLP / Unpublished

(44) Carney, E.W., Quast, J.F., and Zablotny, C.L. (2005) 2

- Methyl

- 2

- Aminopropanol

Hydrochloride: A Reproductive / Developmental Toxicity Screening Test in CD Rats.

The Dow Chemical Company Report No: DR

- 0038

- 3186

- 001 GLP / Unpublished

(45) Carney, E.W. and Thorsrud, B. (2006) AMP: Dermal Developmental Toxicity Study in

CRL:CD(SD) Rats. The Dow Chemical Company Report No: DR

- 0399

- 3940

- 001 GLP / Unpublished

(46) Carney, E.W., Tornesi, B., and Yano, B.L. (2005) AMP

- HCl: Dermal Developmental

Toxicity Probe Study in CD Rats. The Dow Chemical Company Report No: 051051 GLP, Unpublished

(47) Stott, W.T. and Kan, H.L. (2006) 2

- Methyl

- 2

- aminopropanol (AMP): Effects Upon Chol ine

Uptake in Chinese Hamster Ovary Cells. The Dow Chemical Company Report No: DR

- 0038

- 3186

- 002 Not GLP / Unpublished

## 9. References Id 124

- 68

- 5

Date 30.11.2006

88 / 89 (48) Stott, W.T., Zablotny, C.L., and Thomas, J. (2006) Potential of AMP

- HCl to Alter Hepatic

Choline and Phospholipid Synthesis in Preg nant CRL:CD(SD) Rats When Administered by the Dietary Route. The Dow Chemical Company Report No: 050541 GLP, Unpublished

(49) Cipolla et.al. (1997) Airborne Contact Dermatitis from 2

- Amino

- 2

- methyl

- 1

- propanol in a

Cosmetic Company. Arh. hig. rada. tok sikol 48:205

- 209.

Not GLP / Published

## 10. Summary and Evaluation Id 124

- 68

- 5

Date 30.11.2006

89 / 89 10.1 END POINT SUMMARY

10.2 HAZARD SUMMARY

10.3 RISK ASSESSMENT