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 46
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CONTENTS

CONTENTS....................................................................................................................................2

STUDY DIRECTOR STATEMENT OF GLP COMPLIANCE ....................................................4

QUALITY ASSURANCE STATEMENT......................................................................................5

SUMMARY ....................................................................................................................................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 Statement of Data Confidentiality Claim ................................................................8

2 INTRODUCTION AND PURPOSE.....................................................................................9

2.1 Guidelines / Regulations..........................................................................................9

3 TEST ITEM .........................................................................................................................10

4 MATERIALS AND METHODS.........................................................................................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 RESULTS ............................................................................................................................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...............................................................................................................................18
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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 ..........................................46


Harlan Study Number: 41402888 Report 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 Weight. All animals showed expected gains in body weight.

Necropsy. No abnormalities were noted at necropsy.

Conclusion

The acute oral median lethal dose (LD50) of PP-R-001 (CAS# ) in the female 

Wistar strain rat was estimated to be greater than 2000 mg/kg body weight (Globally 

Harmonized 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, Labelling and Packaging of 

Substances and Mixtures.
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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 Procedures) Act 1986 Amendment Regulations 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 Principles 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 

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

decomposed material will no longer be representative of the test sample. 
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1 PREFACE

1.1 Study Information

The study will be performed on the request of Akzo Nobel 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 registration 

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 study. 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.
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2 INTRODUCTION AND PURPOSE

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 tris 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 2002
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3 TEST ITEM

Information as provided by the Sponsor. A Certificate of Analysis supplied by the Sponsor is 

given in Appendix 3.

Identification:

e

CAS number:

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 METHODS

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 period 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 immediately 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, drinking 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.
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The temperature and relative humidity were set to achieve limits of 19 to 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 provided 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 the starting dose.

Groups of fasted animals were treated as follows:

Dose Level

(mg/kg) Specific Gravity Dose Volume

(mL/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.
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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 (LD50) 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, Labelling and Packaging of Substances and Mixtures.
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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 Weight

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 (LD50) of PP-R-001 (CAS# in the female 

Wistar strain rat was estimated to be greater than 2000 mg/kg body weight (Globally 

Harmonized 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, Labelling and Packaging of 

Substances and Mixtures.
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7 REFERENCES

ENVIRONMENT DIRECTORATE, 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 Experimental Animals Used in 

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

and Assessment.

The Animals (Scientific Procedures) Act 1986 Amendment Regulations 2012.

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

Evaluation Studies.
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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

Female 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

Female 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 toxicity

 
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Table 2 Individual Body Weights and Body Weight Changes 

Dose Level

mg/kg

Animal Number 

and Sex

Body Weight (g) at Day Body Weight Gain (g) During Week

0 7 14 1 2

2000

1-0 Female 169 184 192 15 8

1-1 Female 183 216 227 33 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 11
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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 detected
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APPENDICES

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 animals

2-3

2-3 0-1

0-1 2-3 0-1 2-3 0-1 2-3 0-1

0

2-3 0-1 2-3 0-1 2-3 0-1 0

Category 5

> 2000 - 5000

Category 1

> 0 - 5 GHS Category 4> 300 - 2000

Category 3

> 50 - 300

Category 2

> 5 -50

Category 5 or 

unclassified

other other 3 (at 300) 

at 1st step

other 3 (at 2000) 

at 1st step

2 (at 2000) 

at 1st step

1 0 3 (at 50) at 

1st step

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

- 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 harmonised Classification System (mg/kg b.w.)

- Infinity: unclassified

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

b.w.: bodyweight

LD50 cutoff mg/kg 

b.w.
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Appendix 2 Copy of Study Plan 
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 2 (continued) Copy of Study Plan
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Appendix 3 Certificate of Analysis 
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Appendix 4 Monitoring Authority Statement of GLP Compliance