## REPORT

PP

- R

- 001 (CAS# :

Acute Oral Toxicity in the Rat

– Acute Toxic Class Method

Study Director : A Sanders

Test Facility : Harlan Laboratories Ltd.

Shardlow Business Park

Shardlow

Derbyshire

DE72 2GD

UK

Sponsor: Akzo Nobel Functional Chemicals bv

Stationsstraat 77, 3811 MH

P.O. Box 247

3800 AE Amersfoort

THE NETHERLANDS

Study Monitor: Raj Krishnaraj, DVM, PhD, DABT

Akzo Nobel Services Inc.

Phone: 312

- 544

- 7162

raj.krishnaraj@akzonobel.com

Harlan Study Number : 41402888

Study Completion Date: 26 January 2015

Page 1 of 46Harlan Study Number: 41402888 R eport Page 2



## CONTENTS

CONTENTS

................................ ................................ ................................ ................................ .... 2

STUDY DIRECTOR STATE MENT OF GLP COMPLIAN CE

................................ .................... 4

QUALITY ASSURANCE ST ATEMENT

................................ ................................ ...................... 5

SU MMARY

................................ ................................ ................................ ................................ .... 6

Introduction

................................ ................................ ................................ ............................ 6

Methods

................................ ................................ ................................ ................................ . 6

Results

................................ ................................ ................................ ................................ . 6

Conclusion

................................ ................................ ................................ ............................. 6

GENERAL INFORMATION

................................ ................................ ................................ ......... 7

Schedule

................................ ................................ ................................ ................................ . 7

Animal Welfare

................................ ................................ ................................ ...................... 7

Deviations from Study Plan

................................ ................................ ................................ ... 7

Archiving

................................ ................................ ................................ ............................... 7

1 PREFACE

................................ ................................ ................................ .............................. 8

1.1 Study Information

................................ ................................ ................................ .... 8

1.2 Stat ement of Data Confidentiality Claim

................................ ................................ 8

2 INTRODUCTION AND PUR POSE

................................ ................................ ..................... 9

2. 1 Guidelines / Regulations

................................ ................................ .......................... 9

3 TEST ITEM

................................ ................................ ................................ ......................... 10

4 MATERIALS AND METHOD S

................................ ................................ ......................... 10

4.1 Test System

................................ ................................ ................................ ............ 10

4.1.1 Animals and Animal Husbandry

................................ ............................. 10

4.1.2 Justification

................................ ................................ .............................. 11

4.2 Test Item Formulation and Experimental Preparation

................................ .......... 11

4.3 Procedure

................................ ................................ ................................ ............... 11

4.4 Evaluation of Data

................................ ................................ ................................ . 12

5 RESU LTS

................................ ................................ ................................ ............................ 13

5.1 Mortality

................................ ................................ ................................ ................ 13

5.2 Clinical Observations

................................ ................................ ............................ 13

5. 3 Body Weight

................................ ................................ ................................ .......... 13

5.4 Necropsy

................................ ................................ ................................ ................ 13

6 CONCLUSION

................................ ................................ ................................ .................... 13

7 REFERENCES

................................ ................................ ................................ .................... 14

TABLES

................................ ................................ ................................ ................................ ........ 15

APPENDICES

................................ ................................ ................................ ............................... 18Harlan Study Number: 41402888 R eport Page 3



## LIST OF TABLES

Table 1 Individual Clinical Observations and Mortality Data

................................ ................. 15

Table 2 Individual Body Weights and Body Weight Changes

................................ ................ 16

Table 3 Individual Necropsy Findings

................................ ................................ ..................... 17



## LIST OF APPENDICES

Appendix 1 Test Procedure with a Starting Dose of 2000 mg/kg Body Weight

...................... 18

Appendix 2 Copy of Study Plan

................................ ................................ ................................ 19

Appendix 3 Certificate of Analysis

................................ ................................ ........................... 45

Appendix 4 Monitoring Authority Statement of GLP Compliance

................................ .......... 46Harlan Study Number: 41402888 R eport Page 6



## SUMMARY

Introduction

The study was performed to assess the acute oral toxicity of PP

- R

- 001 (CAS# ) in

the Wistar strain rat.



### Methods

A group of three fasted female Wistar (RccHanTM:WIST) strain rats was treated with

PP

- R

- 001 (CAS# at a dose level of 2000 mg/kg body weight. This was followed

by a further group of three fasted females at the same dose level. Dosing was performed

sequentially.

The test item was administered orally undiluted. Clinical signs and body weight development

were monitored during the study. All animals were subjected to gross necropsy.

Results

Mortality. There were no deaths.

Clinical Observations. There were no signs of systemic toxicity.

Body W eight. All animals showed expected gains in body weight.

Necropsy. No abnormalities were noted at necropsy.

Conclusion

The ac ute oral median lethal dose (LD 50 ) of PP

- R

- 001 (CAS# ) in the female

Wistar strain rat was es timated to be greater than 2 0 00 mg/kg body weight (Globally

Har moni z ed Classification System

- Unclassified).

PP

- R

- 001 (CAS# does not meet the criteria for classification according to

Regulation (EC) No 1272/2008, relating to the Classificati on, Labe l ling and Packaging of

Substances and Mixtures.Harlan Study Number: 41402888 R eport Page 7



## GENERAL INFORMATION

Schedule

Experimental Starting Date: 11 November 2014

Experimental Completion Date: 02 December 2014



### Animal Welfare

The study was designed and conducted to cause the minimum suffering or distress to the animals

consistent with the scientific objectives and in accordance with the Harlan Laboratories Ltd,

Shardlow, UK policy on animal welfare and the requirements of the United Kingdom’s Animals

(Scientific Procedure s ) Act 1986 A mendment Regulation s 2012 . The conduct of the study may

be reviewed, as part of the Harlan Laboratories Ltd, Shardlow, UK Ethical Review Process.

The study was conducted in accordance with the UK Home Office Guidance document on

Regulatory Toxicology and Safety Evaluation Studies and the OECD guidance document on

recognition, assessment and use of clinical signs as humane endpoints for experimental animals

used in safety evaluation.



### Deviations from Study Plan

There were no deviations (unplanned changes) from the study plan.



### Archiving

Unless instructed otherwise by the Sponsor, the study plan, all raw data (paper and electronic)

and the final report will be retained in the Harlan Laboratories Ltd., Shardlow, UK archives for

ten years after which instructions will be sought as to further retention or disposal. Further

retention or return of the data will be chargeable to the Sponsor.

No data will be discarded without contacting the Sponsor to obtain their written consent.

According to OECD Princ iples on Good Laboratory Practice (ENV/MC/CHEM(98)17) samples

of test and reference items and specimens should be retained only as long as the quality of the

preparation permits evaluation. Due to inherent properties of peroxides, retention past the

expir ation date is not recommended due to possible decomposition, in which case the resulting

decomposed material will no longer be representative of the test sample.Harlan Study Number: 41402888 R eport Page 8



## 1 PREFACE

1.1 Study Information

The study will be performed on the request of Akzo N obel Functional Chemicals, b.v. No other

party will have any right to use, copy or reproduce the study report for any purpose.

The study monitor will be Raj Krishnaraj, DVM, MSc, PhD., DABT.



### 1.2 Statement of Data Confidentiality Claim

This document holds information that is confidential and trade secret to Akzo Nobel Functional

Chemicals, b.v. It should not be reproduced or microfilmed. It should not be disclosed in any

form to an outside party, nor should information contained herein be used by a regist ration

authority to support registration of any product or substance without the written permission of

Akzo Nobel Functional Chemicals, b.v. or by a trustee duly appointed by Akzo Nobel Functional

Chemicals, b.v. to issue such a letter or access to this st udy. Submission of this information in

the framework of any registration scheme, including REACH, does not constitute a waiver of

any right to confidentiality which may exist under regulatory statute or in any country.Harlan Study Number: 41402888 R eport Page 9



## 2 INTRODUCTION AND PUR POSE

The study was performed to assess the acute oral toxicity of PP

- R

- 001 (CAS# ) in

the Wistar strain rat.



### 2.1 Guidelines / Regulations

This study was designed to be compatible with the procedures indicated by the following

internationally accepted guidelines and recommendations:

• OECD Guideline for the Testing of Chemicals No. 423 “Acute Oral Toxicity

– Acute

Toxic Class Method” (adopted 17 December 2001)

• Method B1 tr is Acute Toxicity (Oral) of Commission Regulation (EC) No. 440/2008

• Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines

for Toxicology Studies, 12 NohSan No. 8147, amended 10 December 2002Harlan Study Number: 41402888 R eport Page 10



## 3 TEST ITEM

Information as p rovided by the Sponsor. A Certificate of Analysis supplied by t he Sponsor is

given in Appendix 3

.

Identification:

e

CAS n u mber:

Batch : BYK004856

- 98

Purity: not applicable

Physical state / Appearance: clear, colorless liquid

Expiry date: 03 April 2017

Storage Conditions: approximately 4 °C in the dark



## 4 MATERIALS AND METHOD S

4.1 Test System

4.1.1 Animals and Animal Husbandry

Female Wistar (RccHanTM:WIST) strain rats were supplied by Harlan Laboratories UK Ltd.,

Oxon, UK. On receipt the animals were randomly allocated to cages. The females were

nulliparous and non

- pregnant. After an acclimatization perio d of at least five days the animals

were selected at random and given a number unique within the study by indelible ink

- marking on

the tail and a number written on a cage card. At the start of the study the animals were eight to

twelve weeks of age. The body weight variation did not exceed ±20% of the mean body weight

of the initially dosed group

.

The animals were housed in groups of three in suspended solid

- floor polypropylene cages

furnished with woodflakes. With the exception of an overnight fast imm ediately before dosing

and for approximately three to four hours after dosing, free access to mains drinking water and

food (2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK)

was allowed throughout the study. The diet, dri nking water and bedding were routinely analyzed

and were considered not to contain any contaminants that would reasonably be expected to affect

the purpose or integrity of the study.Harlan Study Number: 41402888 R eport Page 11

The temperature and relative humidity were set to achieve limits of 19 t o 25 °C and 30 to 70%

respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was

controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve

hours darkness.

The animals were provide d with environmental enrichment items which were considered not to

contain any contaminant of a level that might have affected the purpose or integrity of the study.

4.1.2 Justification

Rats are the preferred species of choice as historically used for safety evaluation studies and are

specified in the appropriate test guidelines.



### 4.2 Test Item Formulation and Experimental Preparation

For the purpose of the study

, PP

- R

- 001 (CAS# ) was used as supplied. The

specific gravity was determined and used to calculate the appropriate dose volume for the

required dose level.



### 4.3 Procedure

Using available information on the toxicity of PP

- R

- 001 (CAS# )

, 2000 mg/kg was

chosen as t he starting dose.

Groups of fasted animals were treated as follows:

Dose Level

(mg/kg) Specific Gravity Dose Volume

(m L /kg) Number of Rats

Female

2000 0.882 2.27 3

2000 0.882 2.27 3

All animals were dosed once only by gavage, using a metal cannula attached to a graduated

syringe. The volume administered to each animal was calculated according to the fasted body

weight at the time of dosing. Treatment of animals was sequential. Sufficient time was allowed

between each group to confirm the survival of the previously dosed animals.

The animals were observed for deaths or overt signs of toxicity 1⁄2, 1, 2 and 4 hours after dosing

and subsequently once daily for fourteen days.Harlan Study Number: 41402888 R eport Page 12

Individual body weights were recorded prior to dosing and seven and fourteen days after

treatment.

At the end of the observation period the animals were killed by cervical dislocation. All animals

were subjected to gross pathological examination. This consisted of an external examination and

opening of the abdominal and thoracic cavities for examination of major organs. The appearance

of any macroscopic abnormalities was recorded. No tissues were retained.



### 4.4 Evaluation of Data

Data evaluations included the relationship, if any, between the exposure of the animal to

PP

- R

- 001 (CAS ) and the incidence and severity of all abnormalities including

behavioral and clinical observations, gross lesions, body weight changes, mortality and any other

toxicological effects.

Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD 50 ) of

PP

- R

- 001 (CAS# was made as shown in the schematic diagram in Appendix 1.

The results were also evaluated according to Regulation (EC) No 1272/2008, relating to the

Classification, Labe l ling and P ackaging of Substances and Mixtures.Harlan Study Number: 41402888 R eport Page 13



## 5 RESULTS

Individual clinical observations and mortality data are given in Table 1

.



### 5.1 Mortality

There were no deaths.



### 5.2 Clinical Observations

No signs of systemic toxicity were noted during the observation period.



### 5.3 Body W eight

Individual body weights and body weight changes are given in Table 2

.

All animals showed expected gains in body weight over the observation period.



### 5.4 Necropsy

Individual necropsy findings are given in Table 3

.

No abnormalities were noted at necropsy.



## 6 CONCLUSION

The acute oral median lethal dose (LD 50 ) of PP

- R

- 001 (CAS# in the female

Wistar strain rat was es timated to be greater than 2 0 00 mg/kg body weight (Globally

Har moni z ed Classification System

- Unclassified ).

PP

- R

- 001 (CAS# does not meet the criteria for classification according to

Regulation (EC) No 1272/2008, relating to the Classification, Labe l ling and Packaging of

Substances and Mixtures.Harlan Study Number: 41402888 R eport Page 14



## 7 REFERENCES

ENVIRONMENT DIRECTORAT E, ORGANISATION FOR ECONOMIC CO

- OPERATION

AND DEVELOPMENT OECD (2000) No. 19 Guidance Document on the Recognition,

Assessment and use of Clinical Signs as Humane Endpoints for Expe rimental Animals Used in

Safety Evaluation . Paris: OECD Environmental Health and Safety Publications Series on Testing

and Assessment.

The Animal s (Scientific Proc edures) Act 1986 A mendment Regulations 2012.

UK HOME OFFICE (2005) Guidance on the Conduct of Regulatory Toxicology and Safety

Evaluation Studies.Harlan Study Number: 41402888 Report Page 15



## TABLES

Table 1 Individual Clinical Observations and Mortality Data

Dose Level

mg/kg Animal

Number

and Sex Effects Noted After Dosing

(Hours) Effects Noted During Period After Dosing

(Days)

1⁄2 1 2 4 1 2 3 4 5 6 7 8 9 10 11 12 13 14

2000 1

- 0

Female 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1

- 1

Fe male 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1

- 2

Female 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

2

- 0

F e male 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

2

- 1

Female 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

2

- 2

Female 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

0 = No signs of systemic toxicityHarlan Study Number: 41402888 Report Page 16

Table 2 Individual Body W eights and Body W eight Changes

Dose Level

mg/kg Animal Number

and Sex Body W eight (g) at Day Body W eight Gain (g) During Week

0 7 14 1 2

2000 1

- 0 Female 169 184 192 15 8

1

- 1 Female 183 216 227 3 3 11

1

- 2 Female 184 205 218 21 13

2

- 0 Female 165 196 211 31 15

2

- 1 Female 161 177 189 16 12

2

- 2 Female 151 164 175 13 11Harlan Study Number: 41402888 Report Page 17

Table 3 Individual Necropsy Findings

Dose Level

mg/kg Animal Number

and Sex Time of Death Macroscopic Observations

2000 1

- 0 Female Killed Day 14 No abnormalities detected

1

- 1 Female Killed Day 14 No abnormalities detected

1

- 2 Female Killed Day 14 No abnormalities detected

2

- 0 Female Killed Day 14 No abnormalities detected

2

- 1 Female Killed Day 14 No abnormalities detected

2

- 2 Female Killed Day 14 No abnormalities detectedHarlan Study Number: 41402888 Report Page 18



## APPENDI CES

Appendix 1 Test Procedure with a Starting Dose of 2000 mg/kg Body Weight

Start

2000 mg/kg

3 animals 300 mg/kg

3 animals 50 mg/kg

3 animals 5 mg/kg

3 animals

2000 mg/kg

3 animals 300 mg/kg

3 animals 50 mg/kg

3 animals 5 mg/kg

3 anim als 2

- 3

0

- 1 2

- 3 0

- 1 2

- 3 0

- 1 2

- 3 0

- 1 2

- 3 0

- 1

0

0

- 1 2

- 3 0

- 1 2

- 3 0

- 1 2

- 3 0

Category 5

> 2000

- 5000 Category 1

> 0

- 5 GHS Category 4

> 300

- 2000 Category 3

> 50

- 300 Cat egory 2

> 5

- 50 Category 5 or

unclassified

other other 3 (at 300)

at 1 st step other 3 (at 2000)

at 1 st step 2 (at 2000)

at 1 st step 1 0 3 (at 50) at

1 st step

50 200 300 500 1000 2000 2500 5000 Infinity 30 25 5

- per step three animals of a single sex (normally females) are used

- 0, 1, 2, 3: Number of moribund or dead animals at each step

- GHS: Globally harmon ised Classification System (mg/kg b.w.) - Infinity : unclassified

- testing at 5000 mg/kg b.w.: see appendix 5

b.w.: bodyweight LD50 cut

off mg/kg

b.w.Harlan Study Number: 41402888 Report Page 19

Appendix 2 Copy of Study PlanHarlan Study Number: 41402888 Report Page 20

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 21

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 22

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 23

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 24

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 25

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 26

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 27

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 28

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 29

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 30

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 31

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 32

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 33

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 34

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 35

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 36

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 37

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 38

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 39

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 40

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 41

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 42

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 43

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 44

Appendix 2 (continued) Copy of Study PlanHarlan Study Number: 41402888 Report Page 45

Appendix 3 Certificate of AnalysisHarlan Study Number: 41402888 Report Page 46

Appendix 4 Monitoring Authority Statement of GLP Compliance