TITLE PAGE FINAL REPORT

Acute oral toxicity study in rats TEST GUIDELINE(S) OECD 423 Commission Regulation (EC) No 440/2008 US EPA OPPTS 870.1100 Japan MAFF 8147 STUDY COMPLETION DATE April 17 2013 SPONSOR

TEST FACILITY

AUTHOR(S)

TEST FACILITY PROJECT IDENTIFICATION

Project No.: 13-BF-OT010 PROJECT IDENTIFICATION

Page 01 of 24 PAGETLE PAGE

This document is directed to USEPA pursuant the Toxic Substances Control Act (TSCA). This document is the property of the sponsor and may be used only for that purpose intended by the sponsor and as authorized under applicable law. Every other or additional use, exploitation, reproduction, publication or submission to other parties requires the written permission of the sponsor, except as otherwise required by applicable law (e.g., TSCA). The use of these studies/documents for regulatory, registration or other commercial purposes [including but not limited to Registration, Evaluation, Authorization and Restriction of Chemical Substances (REACH)], without the express written permission of the sponsor is unauthorized and strictly prohibited.P age 2 of 24

TH IS PAGE IS INTENTIONALLY LEFT BLANKP age 3 of 24

GL P COMPLIANCE STATEMENT
This study was conducted in accordance with the OECD Principles of Good Laboratory Practice and the GLP Principles of the German "Chemikaliengesetz" in the current edition (Chemicals Act), which meet the United States Environmental Protection Agency Good Laboratory Practice Standards [40 CFR Part 160 (FIFRA) and Part 792 (TSCA)], with the exception that recognized differences exist between the GLP Principles/Standards of OECD and the Principles/Standards of FIFRA and TSCA.

Ty ped name of Study Director:
T yped name of Laboratory:

To be filled for USA EPA submission only:

Sponsor:

_______ Date : Typed name of Sponsor: Typed Name of Company:

Submitter:

_______ Date : Typed name of Submitter: Typed Name of Company:P age 4 of 24

Th e following statement is for US EPA submissions only:

FLAGGING CRITERIA
I have applied the criteria of 40 CFR 158.34/40 CFR161.34 for flagging studies for potential adverse effects to the results of the attached study. This study neither meets nor exceeds any of the applicable criteria. meets or exceeds the criteria numbered X

.

Submitter: _______ Date: Typed Name of Signer: Typed Name of Company:

St atement for US EPA submissions only:

FLAGGING CRITERIA The flagging criteria of 40 CFR 158.34/40 CFR161.34 are not applicable for this study type.Pa ge 5 of 24

STA TEMENT OF THE QUALITY ASSURANCE UNIT Title of the report:

Acute oral toxicity study in rats Study director: Date of the final report: April 17, 2013
The study was inspected regularly in accordance with the principles of Good Laboratory Practice for the testing of chemicals as specified by national (ChemG, in its current version) and international (OECD Principles on GLP as revised in 1997 (ENV/MC/CHEM(98)17) standards. The conduct of this study was inspected at intervals to assure the integrity of the study. Inspections were performed according to the testing facility’s Quality Assurance program (i.e., the standard operating procedures of the Quality Assurance Unit). The final report was audited in detail against the approved study protocol and all pertinent raw data. The dates of all study inspections and of the final report as well the dates on which the respective findings were reported to management and to the study director are shown below. Phase of study Inspections Reports Study plan: February 25, 2013 February 25, 2013 Inlife phase March 19, 2013 March 19, 2013 Amendment to the plan: - - Study report: March 27, 2013 March 27, 2013Pa ge 6 of 24

GLP CERTIFICATE (FROM THE COMPETENT AUTHORITY)P age 7 of 24

SI GNATURE PAGE

St udy Director:P age 8 of 24

CO NTRIBUTORS TO THE STUDY / SUPERVISORY LABORATORY PERSONNEL

Head of / Management: Study director: Pathology: Quality Assurance Unit (QAU): Acute Toxicology (Conduction):P age 9 of 24

CO NTENTS
TITLE PAGE 1 THIS PAGE IS INTENTIONALLY LEFT BLANK 2 GLP COMPLIANCE STATEMENT 3 STATEMENT OF THE QUALITY ASSURANCE UNIT 5 CONTRIBUTORS TO THE STUDY / SUPERVISORY LABORATORY PERSONNEL 8 CONTENTS 9 1. SUMMARY 11 2. INTRODUCTION 12 2.1. OBJECTIVES 12 2.2. SELECTION OF DOSES/CONCENTRATIONS 12 2.3. TEST GUIDELINES 12 2.4. TIME SCHEDULE 13 2.5. RETENTION OF RECORDS 13 2.6. ANIMAL WELFARE 13 3. MATERIALS AND METHODS 14 3.1. TEST ITEM 14 3.2. TEST SYSTEMS 14 3.3. HOUSING AND DIET 15 3.4. TEST GROUPS AND DOSES/CONCENTRATIONS 16 3.5. TEST ITEM PREPARATION 16 3.6. ANALYSES 16 3.7. EXPERIMENTAL PROCEDURE 17 3.8. EVALUATION OF RESULTS 18 4. RESULTS 18 4.1. ANALYSES 18P age 10 of 24

4.2. CLINICAL OBSERVATIONS 18 4.2.1. Mortality 18 4.2.2. Clinical signs 18 4.2.3. Body weights 19 4.3. PATHOLOGY 19 5. CONCLUSION 19 APPENDIX I 20 MORTALITY 21 MAXIMUM INCIDENCE OF CLINICAL SIGNS 21 BODY WEIGHTS CHANGES 22 GROSS PATHOLOGY 22 APPENDIX II 23 LIST OF ABBREVIATIONS 24Pa ge 11 of 24



### 1. SUMMARY In an acute oral toxicity study performed according to the Acute Toxic Class method, 2000 mg/kg bw of the test item (preparation in corn oil Ph.Eur.) were administered to two test groups of three fasted Wistar rats by gavage. The following test substance-related clinical observations were recorded on the day of application only: 2000 mg/kg (first test group):

• No mortality occurred

• Impaired general state in all animals

• Piloerection in all animals

• Cowering position in all animals

• Diarrhea in one out of three animals
2000 mg/kg (second test group):

• No mortality occurred

• Impaired general state in one out of three animals

• Piloerection in one out of three animals
The mean body weight increased within the normal range throughout the study period. (with exception of one animal: the weight stagnated in the second observation week). There were no macroscopic pathological findings in the animals sacrificed at the end of the observation period.

The acute oral LD 50 was calculated to be
LD 50 , oral, rat > 2000 mg/kg bwP age 12 of 24



### 2. INTRODUCTION 2.1. OBJECTIVES T he objective of this study was to assess the acute oral median lethal dose following a single oral application to female Wistar rats followed by a 14 day observation period. The study was performed in rats, because this species has been shown to be suitable for this type of study and is recommended by the test guidelines.

2.2. SELECTION OF DOSES/CONCENTRATIONS By request of the sponsor a starting dose of 2000 mg/kg bw was chosen in the first step with 3 female animals. Because no mortality occurred, 2000 mg/kg bw were administered to another group of 3 female animals in the second step.

2.3. TEST GUIDELINES The study was conducted according to the following test guidelines:

- OECD Guideline for Testing of Chemicals No. 423, December 17, 2001 (“Acute Oral T oxicity – Acute Toxic Class Method”)

- Commission Regulation (EC) No 440/2008 of 30 May 2008 laying down test m ethods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorization and Restriction of Chemicals (REACH), Part B: Methods for the determination of toxicity and other health effects: Acute Oral Toxicity – Acute Toxic Class Method; Official Journal of the European Union, No. L 142

- U. S. EPA Health Effects Test Guidelines OPPTS 870.1100, December 2002 ( “Acute Oral Toxicity”)

- Japan MAFF Testing Guideline of 12 Nosan No. 8147 as this in line with OECD 423.Pa ge 13 of 24

2.4 . TIME SCHEDULE Test dates: Study initiation date

(date of signature of the study plan) February 26, 2013
Arrival of the animals and start of acclimatization period (at least 5 days before administration) Experimental starting date (selection of animals for the first treatment within a study) March 04, 2013

First administration March 05, 2013 Day of last observation March 26, 2013 Experimental completion date (documentation of check of raw data) March 27, 2013

2.5. RETENTION OF RECORDS The stability of the test item in the vehicle will be determined indirectly by concentration control analysis. For this purpose, the samples taken were subsequently deep-frozen and were sent to the sponsor for analysis. Additionally retained samples of the test item formulations were stored at at -20°C until finalization of the report and will be destroyed thereafter if no further analytical stability data are requested by the sponsor. At the end of the study (after finalization of the study report), GLP - relevant records and materials, the study plan and the final report will be shipped to the GLP archives of the sponsor:

, and archived for at least the period of time specified in the GLP principles. Copies of raw data and the final report are archived by

Details concerning responsibilities or locations of archiving can be seen from the respective SOPs of the above mentioned archiving locations.

2.6. ANIMAL WELFARE This study was performed in accordance with the German Animal Welfare Act and the European Council Directive 86/609/EEC.Pa ge 14 of 24



### 3. MATERIALS AND METHODS 3.1. TEST ITEM An alysis of the test item was carried out at

. Information on stability or characterization of the test item, which appropriately defines the test batch, is under the responsibility of the sponsor.
Name of test item: Test item No.: 12/0275-1 Batch identification: 0008043725 Purity: 93.8 area-% determined by GC with FID-analysis (for details see non-GLP analytical report No.: AU124464-1 provided by the sponsor.) Homogeneity: The test item was homogeneous by visual inspection. Storage stability: The stability of the test item under storage conditions over the test period was guaranteed by the sponsor, and the sponsor holds this responsibility. Storage conditions: Room temperature Physical state / color: Solid, melt / white Expiry date: December 2013

3.2. TEST SYSTEMS Test species / strain / quality: Rat / Wistar / Crl:WI (Han) SPF

Reasons for selection of the test species: Rats were selected since this rodent species is recommended in the respective test guidelines. Wistar rats were selected since there is extensive experience available in the laboratory with this strain of rats. Age on day 0: Young adult animals (female animals approx. 10 weeks)Pa ge 15 of 24

Sex : As suggested by the OECD guideline nulliparous and non-pregnant female animals were used for the test, because there is no indication that male animals are likely to be more sensitive to the acute effects of the test item. Supplier: Arrival in the testing facility: Acclimatization period of at least 5 days before the beginning of the experimental phase; during the acclimatization period, the animals were accustomed to the environmental conditions of the study and to the diet. Identification: Individual identification by cage cards and tail marking. Body weight on day 0: Animals of comparable weight (± 20% of the mean weight, actual weights see Appendix I)

3.3. HOUSING AND DIET Room temperature / relative humidity: The animals were housed in fully air-conditioned rooms. Central air-conditioning guaranteed a range of 22°C ± 3°C for temperature and of 30 – 70% for relative humidity. There were no deviations from these ranges, which influenced the results of the study. Air changes per hour Approx. 10 Day / night rhythm: 12 h / 12 h (6.00 a.m. – 6.00 p.m. / 6.00 p.m. – 6.00 a.m.) Type of cage: Makrolon cage, type III Number of animals per cage: Single housing Feeding: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany) Drinking water: Tap water ad libitum Bedding: H 15005-29; Ssniff, Spezialitäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany) Enrichment: NGM E-022; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – AustriaPa ge 16 of 24

3.4 . TEST GROUPS AND DOSES/CONCENTRATIONS
Dose (mg/kg bw)
200 0 2000 Administration
1 2 Administration volume

(mL/kg bw)
10 10 Concentration

(g/100 mL)
20 20 No. of animals

(female)
3 3

3.5. TEST ITEM PREPARATION Test item preparation and homogenization until end of each application period: The test item preparation was heated at approx. 60°C. The test item preparation was produced for each application group shortly before application by stirring with a magnetic stirrer. The preparation was administered lukewarm. Vehicle: Corn oil Ph.Eur. Reason for the selection of the vehicle: Solution in corn oil Ph.Eur.
Form of application: Solution

3.6. ANALYSES Conduct of analyses: A sample of the first concentration of the test item formulation was sent to the sponsor for analysis ( Analytical w o rk will be done under the responsibility of the sponsor under GLP. The analytical report is not part of this study report. Stability of the test item preparation: The stability of the test item in the vehicle will be determined indirectly by concentration control analysis. For this purpose, the sample taken was stored at room temperature over the maximum duration of the administration period, subsequently deep-frozen and sent to the sponsor for analysis. Homogeneity of the test item preparation: The homogeneity of the test item preparation in the vehicle was not determined by analysis because the test item preparation was a solution.Pa ge 17 of 24

Concentration control analysis of the test item preparation: A concentration control analysis of the test item preparation and archiving of the respective data will be done by the sponsor.
Analysis of feed: The feed used in the study was assayed for chemical and microbial contaminants by the manufacturer in quarterly intervals. Analysis of drinking water: The drinking water was regularly assayed for contaminants by the municipal authorities of . The Drinking Water Regulation of Dec. 5, 1990 served as the guideline for maximum tolerable contaminants. Bedding and enrichment analysis: The bedding and enrichment were regularly assayed for contaminants (chlorinated hydrocarbons and heavy metals).

3.7. EXPERIMENTAL PROCEDURE Route of administration: Single oral administration by gavage. Fasting period: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum. Time of day of administration: In the morning Observation period: 14 days Body weight determination: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation. Clinical observations: Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals. Mortality: A check for any dead or moribund animals was made at least once each workday, these records are archived byP age 18 of 24

Pa thology: Necropsy with gross-pathology examination on the last day of the observation period after sacrifice by CO 2 - i nhalation in a chamber with increasing concentrations over time. Histology: No histological examinations were performed.

3.8. EVALUATION OF RESULTS Calculations were performed using Microsoft Excel 2003 and checked with a calculator.


### 4. RESULTS Tables with the results of the clinical and pathological examinations are presented in t he Appendix I.

4.1. ANALYSES Analysis of the test item preparations will be done by the sponsor and reported separately. Analysis of feed: In view of the aim and duration of the study the contaminants occurring in commercial feed should not influence the results. Analysis of drinking water: In view of the aim and duration of the study there are no special requirements exceeding the specification of drinking water. Analysis of bedding and enrichment: In view of the aim and duration of the study there are no special requirements exceeding the specification of a commercial grade monitored by the manufacturer.

4.2. CLINICAL OBSERVATIONS 4.2.1. Mortality No mortality occurred in both 2000 mg/kg bw test groups.
4.2.2. Clinical signs In all animals of the first 2000 mg/kg bw test group impaired general state and piloerection were observed at hour 1 and persisted in two animals until hour 4 or 5 after administration.Pa ge 19 of 24

Dia rrhea was noted in one animal at hour 0, while cowering position was observed in all animals at hour 1. Clinical signs in the second 2000 mg/kg bw test group revealed impaired general state and piloerection in one animal from hour 0 until hour 2 after administration. In two animals of this test group no clinical signs were observed during clinical examination.
4.2.3. Body weights The mean body weight increased within the normal range throughout the study period. (with exception of one animal: the weight stagnated in the second observation week).

4.3. PATHOLOGY There were no macroscopic pathological findings in the animals sacrificed at the end of the observation period.


### 5. CONCLUSION Under the conditions of this study the median lethal dose of after oral administration was found to be greater than 2000 mg/kg bw in rats.P age 20 of 24



### APPENDIX IP age 21 of 24

MOR TALITY

MA X IMUM INCIDENCE OF CLINICAL SIGNSP age 22 of 24

BO DY WEIGHTS CHANGES

GR OSS PATHOLOGYP age 23 of 24



### APPENDIX IIP age 24 of 24

LI ST OF ABBREVIATIONS a.m. = ante meridiem (before noon) approx. = Approximately bw = body weight CMC = sodium Carboxymethylcellulose D, d = day(s) d0 = day 0 (day of application / day of administration) e.g. exempli gratia (for example) f = Female G; g = Gram h = Hour Kg; kg = Kilogram L = Liter

m = Male mg = Milligram mL = Milliliter mm = Millimeter mw = Mean weight No.; no. = Number p.m. = post meridiem (after noon) SPF = specific pathogen free

v = Volume v/v = volume per volume

w = Weight WE = Week end/ public holydays w/w = weight per weight Symbols: °C = degree Celsius X = affected animals
~ = Circa
= = Analogous % = per cent
- = no abnormality / = no data + = No further data  = Weight at death