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### REPORT

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

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Final 03 January2013Huntingdon 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 Alconbury Huntingdon Cambridgeshire PE28 4HS 

## UK

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# 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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# J List ofTables Huntingdon Life Sciences[

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

## List ofAppendices 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 ofAnnexes Annex 1 Annex2 Certificate ofanalysis...................................................................................24 GLP Compliance Statement.........................................................................26

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# 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 LaboratoryPractice standards and, withthe exceptionnoted below; I consider the data generated to be valid: The UK Good Laboratory Practice Regulations (StatutoryInstrument 1999 No. 3106, as amended by StatutoryInstrument2004 No. 994). OECD Principles ofGood Laboratory Practice (as revised in 1997), ENV /MC/CHEM(98)17. EC Commission Directive 2004/10/EC of 11 February2004 (Official Journal No. L 50/44). Theseprinciples ofGood Laboratory Practice are accepted bythe regulatory authorities of the United States ofAmerica and Japan onthe basis ofintergovernmental agreements. Chemical analysis ofsamples ofthe formulated test substance for determination ofstability, homogeneity and concentration was notundertaken for this study.

David G ColemanBSc (Hans) Study Director Huntingdon Life Sciences

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## QualityAssurance 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 ofInspection

Protocol Audit 07Nov2013 ReportAudit 30 Dec 2013 Date ofReporting to Study Director and Management 07 Nov2013 30 Dec 2013
Process based inspections: At or about the time this studywas inprogress inspections of procedures employed onthis type ofstudy were carried out. These were conducted and reported to appropriate Company Management as indicated below: Process Based Inspections Date(s) oflnspection Date ofReporting to Management Acute studyterminal 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 22Nov 2013 22 Nov 2013 post dose observations
In addition, an inspection ofthe facility where this study was conducted was carried outon an annual basis. These inspections were promptly reported to Company Management.

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# ~ 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 StudyManager

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## Summary The studywas performed to assess the acute oral toxicity of3-Phenylpropyl benzoate a plasticizer/coalescent, to the rat.



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# 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 ofthe following guidelines: EEC Methods for the determination oftoxicity and otherhealth effects. CommissionRegulationNo. 440/2008, Part B, Method B.! tris. Acute Oral Toxicity: Acute Toxic Class Method. 30 May 2008. OECD Guideline for Testing ofChemicals No.423 'Acute Oral Toxicity- Acute Toxic Class Method' Adopted 17 December 2001. EPAHealth Effects Test Guidelines OPPTS 870.1100 Acute Oral Toxicity EPA712-C-02-190. 2002. Japanese Ministry ofAgriculture, Forestry and Fisheries, Test Datafor Registration ofAgricultural 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 ofa single sex per step. Depending onthe mortality and/or the moribund status ofthe animals, on average two to four steps may be necessary to allow ajudgementon the acute toxicity ofthe 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 withthe requirements ofcurrent, internationally recognised Good Laboratory Practice Standards, and the applicable sections ofthe United Kingdom Animals (Scientific Procedures) Act 1986 (Amendment Regulations (2012)). 

## 1.3 Test system The rat was chosen as it has been shownto be a suitable model for this type ofstudy 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 Sponsorthe initial dose was 2000 mg/kg. 

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1.6 Control animals 

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## 2. Experimental Procedure 2.1 Study schedule 2.1.1 Time schedule Study initiation: (Protocol signedby Study Director) Experimental start date: (Animal allocation) Initial day oftreatment: Experimental completion date: (Date oflast necropsy) Study completion: Huntingdon Life Sciences[

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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 Sponsorregarding 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:

113-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-propanolbenzoate 3-PPB 60045-26-3 Plasticizer/coalescent Clear, colourless liquid Room temperature undernitrogen SponsorLot number: Expiry date: Purity: Sample receipt: Huntingdon Life Sciences[_ 

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EH101912A 19 October 2014 99.1% 22 October 2013 The Sponsorwas responsible for the characterisation ofthe test substance and the documentation ofthe methods ofsynthesis, fabrication "or derivation and stability. 

### 2.2.2 Vehicle The vehicle used for the study was corn oil. 

### 2.2.3 Formulation Thetest substance was formulated at a concentration of200 mg/mL inthe vehicle and administered ata volume of 10 mL/kg body weight. The test substance formulation was prepared on the day ofdosing. The absorption ofthe test substance was not determined. Determination ofthe homogeneity, stability and purity ofthe test substance ortest substance formulations were not undertaken as part ofthis study. Detailed records oftest substance usage were maintained. The amount oftest substance necessary to preparethe 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

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### 2.3.3 Administration 

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2.4 Serial observations 2.4.1 Mortality



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# .. 2.4.2 Clinical observations

2.4.3 Body weight 

# [2.5 Necropsy and macropathology 2.5.1 Method of kill 

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# J 2.6 Computer systems The computer systems that were used onthis study to acquire and quantify data include:

System name* System function Xybion Pristima Used for Pharmacy test substance management. 

# • All version numbers ofthe systems are maintained by Huntingdon Life Sciences 

### 2.7 2.7.1 

### Quality Assurance and archiving procedures Quality Assurance Details ofthe Quality Assurance inspections and audits are presented on the Quality Assurance Statement. 

### 2.7.2 Archives Following completion ofthis study all raw data were stored in the archives of HuntingdonLife Sciences.



### 2.8 Deviations from protocol There were no deviations from protocol.

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## 3. Results Huntingdon Life Sciencet 

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3.1 Mortality 

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### Clinical signs Table l 

# - J 3. Body weight A endix l and Appendix 2 

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3.4 Macroscopic pathology Appendix 3 

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## 5. References Huntingdon Life Sciences[ 

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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

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No. ofrats in groups of3 showingsigns Dose (mg!kg)Huntingdon Life Sciences[ 

# J Appendix 1 Individual and group mean body weights (g)

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# J Appendix 2 Individual body weight changes (g) 

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### Appendix 3 Macroscopic findings 

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Annex 1 Certificate of analysis

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## (**Em~ald** 

## Performance MaterialsTM

## ~ Kalama Chemical Huntingdon Life Sciences[ 

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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



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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 42Annex2 

# - Huntingdon Life SciencesL GLP Compliance Statement 

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### HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2012 

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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 Toxlclty Ecosystems

Toxicology Residue Studies ReissuedFebruary2013 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).

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# I,._J•:3 Dr. Andrew J. Gray

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