Huntingdon Life Sciences 

Working for a better future 

REPORT 

Seu.~~ej ~ ~~ 

ft~ Oo cU 

3-Phenylpropyl benzoate: Acute Oral Toxicity to the Rat (Acute 

Toxic Class Method) 

~S study numbe" 

Version ID: 

Issue date: 

Page 1 of 27 

-------) 

Final 

03 January 2013 
Huntingdon Life Sciences[ J 

Sponsor and Test Facility Details 

Sponsor 

Test facility 

Emerald Performance Materials 

LLC 

2020 Front Street 

Suite 100 

Cuyahoga Falls 

OH 44221 

USA 

Huntingdon Life Sciences 

Huntingdon Research Centre 

Woolley Road 

Alcon bury 

Huntingdon 

Cambridgeshire 

PE28 4HS 

UK 

2 
Huntingdon Life Sciences[ J 

Table of Contents 

Sponsor and Test Facility Details .......................................................................................... 2 

Compliance with Good Laboratory Practice .......................................................................... 5 

Quality Assurance Statement. ............................................................................................... 6 

Contributing Scientists .......................................................................................................... 7 

Summary .............................................................................................................................. 8 

1. Introduction .................................................................................................................. 9 

1.1 Objective ........................................................................................................... 9 

1.2 Regulatory compliance ..................................................................................... 9 

1.3 Test system ...................................................................................................... 9 

1.4 Route of administration ..................................................................................... 9 

1.5 Treatment groups and doses .......................................................................... 10 

1.6 Control animals ............................................................................................... 10 

2. Experimental Procedure ............................................................................................. 11 

2.1 Study schedule ............................................................................................... 11 

2.1.1 Time schedule ..................................................................................... 11 

2.2 Test substance and formulation ...................................................................... 11 

2.2.1 Test substance .................................................................................... 11 

2.2.2 Vehicle ................................................................................................ 12 

2.2.3 Formulation ......................................................................................... 12 

2.3 Animal management ....................................................................................... 12 

2.3.1 Animal supply, acclimatisation and allocation ...................................... 12 

2.3.2 Animal housing, diet and water supply ................................................ 13 

2.3.3 Administration ..................................................................................... 14 

2.4 Serial observations ......................................................................................... 14 

2.4.1 Mortality .............................................................................................. 14 

2.4.2 Clinical observations ........................................................................... 14 

2.4.3 Body weight ........... :, ........................................................................... 14 

2.5 Necropsy and macropathology ....................................................................... 14 

2.5.1 Method ofkill ....................................................................................... 14 

2.5.2 Macroscopic pathology ........................................................................ 14 

2.6 Computer systems .......................................................................................... 15 

2.7 Quality Assurance and archiving procedures .................................................. 15 

2. 7.1 Quality Assurance ............................................................................... 15 

2. 7.2 Archives .............................................................................................. 15 

2.8 Deviations from protocol ................................................................................. 15 

3. Results ....................................................................................................................... 16 

3. 1 Mortality .......................................................................................................... 16 

3.2 Clinical signs ................................................................................................... 16 

3.3 Body weight.. .................................................................................................. 16 

3.4 Macroscopic pathology ................................................................................... 16 

4. Conclusion ................................................................................................................. 17 

5. References ................................................................................................................. 18 

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List of Tables 

Huntingdon Life Sciences[ 

Table 1 Clinical signs ................................................................................................ 19 

List of Appendices 

Appendix 1 

Appendix 2 

Appendix 3 

Appendix 4 

Individual and group mean body weights (g) ................................................ 20 

Individual body weight changes (g) .............................................................. 21 

Macroscopic findings ................................................................................... 22 

Test procedure ............................................................................................. 23 

List of Annexes 

Annex 1 

Annex2 

Certificate of analysis ................................................................................... 24 

GLP Compliance Statement. ........................................................................ 26 

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Huntingdon Life Sciences{:~ J 

Compliance with Good Laboratory Practice 

3-Phenylpropyl benzoate: Acute Oral Toxicity to the Rat (Acute Toxic 

Class Method) 

The study described in this report was conducted in compliance with the following Good 

Laboratory Practice standards and, with the exception noted below; I consider the data 

generated to be valid: 

The UK Good Laboratory Practice Regulations (Statutory Instrument 1999 

No. 3106, as amended by Statutory Instrument 2004 No. 994). 

OECD Principles of Good Laboratory Practice (as revised in 1997), 

ENV /MC/CHEM(98) 17. 

EC Commission Directive 2004/1 0/EC of 11 February 2004 (Official Journal 

No. L 50/44). 

These principles of Good Laboratory Practice are accepted by the regulatory authorities of 

the United States of America and Japan on the basis of intergovernmental agreements. 

Chemical analysis of samples of the formulated test substance for determination of stability, 

homogeneity and concentration was not undertaken for this study. 

David G Coleman BSc (Hans) 

Study Director 

Huntingdon Life Sciences 

5 
Huntingdon L~e Sciences: [ ] 

Quality Assurance Statement 

3-Phenylpropyl benzoate: Acute Oral Toxicity to the Rat (Acute Toxic 

Class Method) 

The following inspections and audits have been carried out in relation to this study: 

Study Phase Date of Inspection 

Protocol Audit 07Nov2013 

Report Audit 30 Dec 2013 

Date of Reporting 

to Study Director 

and Management 

07 Nov2013 

30 Dec 2013 

Process based inspections: At or about the time this study was in progress inspections of 

procedures employed on this type of study were carried out. These were conducted and 

reported to appropriate Company Management as indicated below: 

Process Based Inspections Date(s) oflnspection Date of Reporting to 

Management 

Acute study terminal 24 Sep 2013 24 Sep 2013 

procedures 

Clinical signs examination 05 Nov 2013 05 Nov 2013 

Formulation procedures- 07Nov2013 07 Nov2013 

Liquid 

Bodyweights 21 Nov 2013 21 Nov 2013 

Dose administration and 22 Nov 2013 22 Nov 2013 

post dose observations 

In addition, an inspection of the facility where this study was conducted was carried out on an 

annual basis. These inspections were promptly reported to Company Management. 

6 
Huntingdon Life Sciences[ ~ 

Contributing Scientists 

3-Phenylpropyl benzoate: Acute Oral Toxicity to the Rat (Acute Toxic 

Class Method) 

Study management 

David G Coleman BSc (Hons) 

Study Director 

Kerstin Pryke BSc (Hons) 

Trainee Study Manager 

7 
Huntingdon Life Sciences:[ ] 

Summary 

The study was performed to assess the acute oral toxicity of3-Phenylpropyl benzoate a 

plasticizer/coalescent, to the rat. 

Conclusion 

[ J 

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Huntingdon Life Sciences[ J 

1. Introduction 

1.1 Objective 

The study was performed to assess the acute oral toxicity of3-Phenylpropyl benzoate a 

plasticizer/coalescent, to the rat. 

1.2 Regulatory compliance 

The study was designed to meet the requirements of the following guidelines: 

EEC Methods for the determination of toxicity and other health effects. 

Commission Regulation No. 440/2008, Part B, Method B.! tris. Acute Oral 

Toxicity: Acute Toxic Class Method. 30 May 2008. 

OECD Guideline for Testing of Chemicals No.423 'Acute Oral Toxicity- Acute 

Toxic Class Method' Adopted 17 December 2001. 

EPA Health Effects Test Guidelines OPPTS 870.1100 Acute Oral Toxicity 

EPA 712-C-02-190. 2002. 

Japanese Ministry of Agriculture, Forestry and Fisheries, Test Data for Registration 

of Agricultural Chemicals, Acute oral toxicity (2-1-1), 12 Nousan No 8147, 

Agricultural Production Bureau, November 24, 2000. 

The acute toxic class method is a stepwise procedure that uses three animals of a single sex 

per step. Depending on the mortality and/or the moribund status of the animals, on average 

two to four steps may be necessary to allow a judgement on the acute toxicity of the test 

substance. The acute toxic class method is based on biometric evaluations and has been 

subjected to international validation (Schlede eta!. 1995). 

The study was conducted in accordance with the requirements of current, internationally 

recognised Good Laboratory Practice Standards, and the applicable sections of the United 

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

1.3 Test system 

The rat was chosen as it has been shown to be a suitable model for this type of study and is 

the species recommended by the test guidelines. As there were no data to indicate males 

were likely to be more sensitive to the test substance, the testing was performed in females 

only. 

1.4 Route of administration 

The rats were dosed by oral gavage as the test substance may be ingested accidentally. 

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1.5 Treatment groups and doses 

Huntingdon Life Sciences[ 

The dose level for the study was chosen in compliance with the study guidelines. Based on 

information provided by the Sponsor the initial dose was 2000 mg/kg. 

---

1.6 Control animals 

[ ·J 

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2. Experimental Procedure 

2.1 Study schedule 

2.1.1 Time schedule 

Study initiation: 

(Protocol signed by Study Director) 

Experimental start date: 

(Animal allocation) 

Initial day of treatment: 

Experimental completion date: 

(Date of last necropsy) 

Study completion: 

Huntingdon Life Sciences[ J 

01 November 2013 

07 November 2013 

13 November 2013 

29 November 2013 

When Study Director signs final report 

2.2 Test substance and formulation 

2.2.1 Test substance 

Information supplied by the Sponsor regarding the test substance is contained in the test 

substance data sheet, which is retained in study records. 

The following information is given in summary: 

Identification: 

Alternative Identities: 

Chemical name: 

Alternative Chemical names: 

CAS registry number: 

Intended use: 

Appearance: 

Storage conditions: 

Supplier: 

11 

3-Phenylpropyl benzoate 

X-613 

K-Fiex 613 

Kalama K-Flex 613 

Kalama K-Flex 613 Plasticizer 

Benzenepropanol, !-benzoate 

!-Propanol, 3-phenyl-, benzoate 

3-Phenyl-l-propanol benzoate 

3-PPB 

60045-26-3 

Plasticizer/coalescent 

Clear, colourless liquid 

Room temperature under nitrogen 

Sponsor 
Lot number: 

Expiry date: 

Purity: 

Sample receipt: 

Huntingdon Life Sciences[_ J 

EH101912A 

19 October 2014 

99.1% 

22 October 2013 

The Sponsor was responsible for the characterisation of the test substance and the 

documentation of the methods of synthesis, fabrication "or derivation and stability. 

2.2.2 Vehicle 

The vehicle used for the study was corn oil. 

2.2.3 Formulation 

The test substance was formulated at a concentration of200 mg/mL in the vehicle and 

administered at a volume of 10 mL/kg body weight. 

The test substance formulation was prepared on the day of dosing. 

The absorption of the test substance was not determined. 

Determination of the homogeneity, stability and purity of the test substance or test substance 

formulations were not undertaken as part of this study. 

Detailed records oftest substance usage were maintained. The amount of test substance 

necessary to prepare the formulations and the amount actually used were determined on each 

occasion. The difference between these amounts was checked before the formulations were 

dispensed. 

2.3 Animal management 

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2.3.2 Animal housing, diet and water supply 

13 

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Huntingdon Life SciencesL . ~ 
2.3.3 Administration 

-

2.4 Serial observations 

2.4.1 Mortality 

c~ . .. 

2.4.2 Clinical observations 

2.4.3 Body weight 

[ 

2.5 Necropsy and macropathology 

2.5.1 Method of kill c-.. .. 

2.5.2 Macroscopic pathology 

., 

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Huntingdon Life Sciences[ J 

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Huntingdon Life Sciences:[ J 

2.6 Computer systems 

The computer systems that were used on this study to acquire and quantify data include: 

System name* System function 

Xybion Pristima Used for Pharmacy test substance management. 

• All version numbers of the systems are maintained by Huntingdon Life Sciences 

2.7 

2.7.1 

Quality Assurance and archiving procedures 

Quality Assurance 

Details of the Quality Assurance inspections and audits are presented on the 

Quality Assurance Statement. 

2.7.2 Archives 

Following completion of this study all raw data were stored in the archives of 

Huntingdon Life Sciences. 

2.8 Deviations from protocol 

There were no deviations from protocol. 

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3. Results 

Huntingdon Life Sciencet J 

3.1 Mortality 

J ·-

L:.2 : ' 

Clinical signs 

Table l 

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J 

3. Body weight 

A endix l and Appendix 2 

---

3.4 Macroscopic pathology 

Appendix 3 

[ 

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5. References 

Huntingdon Life Sciences[ J 

Schlede E, Mischke U, Diener Wand Kayser D (1995). The International Validation Study of 

the Acute-Toxic-Class Method (Oral). Arch Toxicol. 69, 659-670. 

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Table 1 Clinical signs 

Clinical signs 

19 

Huntingdon Life Science[ J 

No. of rats in groups of3 showing signs 

Dose (mg!kg) 
Huntingdon Life Sciences[ J 

Appendix 1 Individual and group mean body weights (g) 

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Huntingdon Life Sciences[ J 

Appendix 2 Individual body weight changes (g) r---

21 
Appendix 3 Macroscopic findings .---...: 

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Huntingdon Life Sciences[ J 

Annex 1 Certificate of analysis 

24 
( Em~ald Performance MaterialsTM 

~ Kalama Chemical 

Huntingdon Life Sciences[ J 

CER11FICATE OF ANALYSIS 

Emerald Kalama Chemical, LLC 

1296 Third Street North>vest, Kalama, WA 98625 

Phone· (360} 673-2550 Fax: (360) 673-3564 

PRODUCT: Kalama® K-Fiex® 613 Plasticizer 

This is to certify that the product identified below has been representatively sampled and analyzed according to 

our quality control programs. It meets or exceeds the s ecifications for K-Fiex 613. 

Lot Number: 

Date of Manufacture: 

CAS # 60045-26-3 

Appearance, visual 

Assay,% 

Acidity,% 

Color, APHA 

"' J 

L---

SPECIFICATIONS 

Clear and free of particulates 

99.0, Minimum 

0.3, Maximum 

100, Maximum 

Certificate computer generated thus not requiring a signature. 

25 

ANALYSIS 

Pass 

99.1 

0.1 

42 
Annex2 

- Huntingdon Life SciencesL 

GLP Compliance Statement 

J 

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Huntingdon Life ScienceL 1-] • 

HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2012 

i nH) Department 

~ ofHealth 

THE DEPARTMENT OF HEALTH OF THE GOVERNMENT 

OF THE UNITED KINGDOM 

GOOD LABORATORY PRACTICE 

STATEMENT OF COMPLIANCE 

IN ACCORDANCE WITH DIRECTIVE 2004191EC 

TEST FACILITY 

Huntingdon Life Sciences 

Woolley Road 

Alconbury 

Huntingdon 

Cambridgeshire 

PE284HS 

TEST TYPE(S) 

Analytical/Clinical Chemistry 

Environmental Fate 

Environmental T oxlclty 

Ecosystems 

Toxicology 

Residue Studies 

Reissued February 2013 to include Residue 

Studies 

DATE OF INSPECTION 20 to 22 November 2012 

An inspection for compliance with the Principles of Good Laboratory Practice was 

carried out at the above test facility as part of the UK Good Laboratory Practice 

Compliance Monitoring Programme. 

This statement confirms that, on the date of issue, the UK" Good Laboratory 

Practice Monitoring Authority were satisfied that the above test facility was 

operating in compliance with the OECD Principles of Good Laboratory Practice. 

This statement constitutes a Good Laboratory Practice Instrument (as defined in the 

UK Good Laboratory Practice Regulations 1999). 

t.:t.. I ,._J •:3 

Head, 

Dr. Andrew 

UK GLP 

J. Gray 

Monitoring Authority 

~ 

~ 

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