RCC STUDY NUMBER B04803 Report Page 2

Ciba EFKA 8530



# TABLE OF CONTENTS

1 PREFACE ..........................................................................................................................3

1.1 GENERAL ................................................................................................................3

1.2 RESPONSIBILITIES .................................................................................................3

1.3 SCHEDULE ..............................................................................................................3

1.4 ARCHIVING ..............................................................................................................4

1.5 SIGNATURE PAGE ..................................................................................................5

1.6 QUALITY ASSURANCE GLP TOXICOLOGY ..........................................................6

1.7 STATEMENT OF COMPLIANCE .............................................................................7

1.8 TEST GUIDELINES ..................................................................................................8

1.9 ANIMAL WELFARE ..................................................................................................8

2 SUMMARY ........................................................................................................................9

3 CONCLUSION ...................................................................................................................9

4 PURPOSE .......................................................................................................................10

5 MATERIALS AND METHODS .........................................................................................10

5.1 TEST SYSTEM .......................................................................................................10

5.2 HUSBANDRY .........................................................................................................10

5.3 TEST ITEM .............................................................................................................11

5.4 VEHICLE ................................................................................................................11

5.5 DOSE FORMULATION ..........................................................................................12

5.6 TREATMENT ..........................................................................................................12

5.7 OBSERVATIONS ...................................................................................................12

6 PATHOLOGY ..................................................................................................................12

6.1 NECROPSY ...........................................................................................................12

7 STATISTICAL ANALYSIS ...............................................................................................13

8 DATA COMPILATION .....................................................................................................13

9 RESULTS ........................................................................................................................14

9.1 MORTALITY ...........................................................................................................14

9.2 CLINICAL SIGNS ...................................................................................................14

9.3 BODY WEIGHTS ....................................................................................................14

9.4 MACROSCOPIC FINDINGS ..................................................................................14

9.5 MEDIAN LETHAL DOSE ........................................................................................14

10 INDIVIDUAL FINDINGS ..................................................................................................15

10.1 MORTALITY / CLINICAL SIGNS ............................................................................15

10.2 BODY WEIGHTS ....................................................................................................16

10.3 MACROSCOPIC FINDINGS ..................................................................................16

11 GLP – CERTIFICATION ..................................................................................................17RCC STUDY NUMBER B04803 Report Page 3

Ciba EFKA 8530



# 1 PREFACE

1.1 GENERAL

Title Ciba EFKA 8530: Acute Oral Toxicity Study in Rats

Sponsor Ciba Specialty Chemicals Inc. Segment Coating Effects

P.O. Box WRO 1045.1.30

CH-4002 Basel / Switzerland

Study Monitor Mr. Henry Su

Test Facility RCC Ltd Toxicology

Wölferstrasse 4

CH-4414 Füllinsdorf / Switzerland



## 1.2 RESPONSIBILITIES

Study Director M. Ott

Deputy for Study Director G. Arcelin

Technical Coordinators F. Frickert / M. Bernstein

Head of RCC Quality Assurance I. Wüthrich



## 1.3 SCHEDULE

Experimental Starting Date 15-NOV-2006

Experimental Completion Date 08-DEC-2006

Delivery of Animals 15-NOV-2006 (females, 2000 mg/kg) 17-NOV-2006 (females, 2000 mg/kg)

Acclimatization 15-NOV-2006 to 21-NOV-2006 (females, 2000 mg/kg) 17-NOV-2006 to 23-NOV-2006 (females, 2000 mg/kg)

Treatment 22-NOV-2006 (females, 2000 mg/kg) 24-NOV-2006 (females, 2000 mg/kg)

Observation 15-NOV -2006 to 06-DEC-2006 (females, 2000 mg/kg) 17-NOV-2006 to 08-DEC-2006 (females, 2000 mg/kg)RCC STUDY NUMBER B04803 Report Page 4

Ciba EFKA 8530



## 1.4 ARCHIVING

RCC Ltd (CH-4452 Itingen / Switzerland) will retain the study plan, raw data, a sample of test item(s) and the final report of the present study for at least ten years. No data will be discarded without the Sponsor's written consent.RCC STUDY NUMBER B04803 Report Page 8

Ciba EFKA 8530



## 1.8 TEST GUIDELINES

The study procedures described in this report meet or exceed the requirements of the following guidelines:

Directive 2004/73/EC, B.1 tris "Acute Oral Toxicity-Acute Toxic Class Method”, April 29,

2004.

OECD Guidelines for the Testing of Chemicals, Number 423 "Acute Oral Toxicity – Acute Toxic Class Method", adopted 17 th 

### December 2001.



## 1.9 ANIMAL WELFARE

This study was performed in an AAALAC-approved laboratory in accordance with the Swiss

Animal Protection Law under license no. 34.RCC STUDY NUMBER B04803 Report Page 9

Ciba EFKA 8530



# 2 SUMMARY

Two groups, each of three female HanRcc:WIST (SPF) rats, were treated with Ciba EFKA 8530 by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was diluted in vehicle (purified water) at a concentration of 0.2 g/mL and administered at a dosing volume of 10 mL/kg.

The animals were examined daily during the a cclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30 minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Mortality/viability was recorded at approximately 30 minutes, 1, 2, 3 and 5 hours after administration on test day 1 (with the c linical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

The following animals were treated and percentage of mortality was observed:

6 females treated at 2000 mg/kg 17%

One animal was killed in extremis for ethical reasons on test day 13.

Slightly to moderately ruffled fur was noted in all animals from the 30-minute, 1-hour and 2- hour reading to test day 2 and persisted in two animals until test day 3 and in one animal until test day 4. Hunched posture was noted in all animals of the second treated group from the 30-minute or 1-hour reading to the 5-hour reading and slight sedation was noted in the same animals from the 2- to the 5-hour reading. The animal killed in extremis for ethical reasons showed slight to moderate ruffled fur and hunched posture from test day 11 to 13, labored respiration from test day 8 to 13 and cyanosis on test day 13.

One animal showed a slight loss of body weight (1.8 %) during the last week of observation. The body weight of the other animals was within the range commonly recorded for this strain and age.

Not collapsed lungs and the disgestif tract (stomach, duodenum, jejunum, ileum, caecum, colon and rectum) distended with gas was noted at the unscheduled necropsy of the animal killed in extremis for ethical reasons. No macroscopic findings were recorded at all other

necropsy.



# 3 CONCLUSION

The median lethal dose of Ciba EFKA 8530 after single oral administration to female rats, observed over a period of 14 days is:

LD 50 (female rat): greater than 2000 mg/kg body weightRCC STUDY NUMBER B04803 Report Page 10

Ciba EFKA 8530



# 4 PURPOSE

The purpose of this study was to assess the acute toxicity of Ciba EFKA 8530 when administered by a single oral gavage to rats, followed by an observation period of 14 days.

This study provides information for both hazard assessment and hazard classification purposes.



# 5 MATERIALS AND METHODS

5.1 TEST SYSTEM

Test system Rat, HanRcc:WIST (SPF)

Rationale Recognized by the international guidelines as a recommended test system.

Source RCC Ltd, Laboratory Animal Services

CH-4414 Füllinsdorf / Switzerland

Number of animals per group 3 females

Total number of animals 6 females

Age when treated 11 weeks

Identification Unique cage number and corresponding color-coded spots

on the tail. The animals were marked at acclimatization start.

Randomization Selected by hand at time of delivery. No computer generated randomization program.

Acclimatization Under laboratory conditi ons, after health examination. Only animals without any visible signs of illness were used for the study.



## 5.2 HUSBANDRY

Room no. 0105 / RCC Ltd, Füllinsdorf

Conditions Standard Laboratory Conditions. Air-conditioned with 10-15air changes per hour, and continuously monitored environment with ranges for room temperature 22 ± 3 °C and for relative humidity between 30-70 % (values above 70 % during cleaning process possible), automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime light period.

Accommodation In groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz/Switzerland).RCC STUDY NUMBER B04803 Report Page 11

Ciba EFKA 8530



### Diet Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 48/06 (Provimi Kliba AG, CH-4303 Kaiseraugst/Switzerland) ad libitum . Results of analyses for

contaminants are archived at RCC Ltd.

Water Community tap water from Füllinsdorf ad libitum . Results ofbacteriological, chemical and contaminant analyses are archived at RCC Ltd.



## 5.3 TEST ITEM

The following information was provided by the sponsor:

Identity Ciba EFKA 8530

Description Yellowish liquid with characteristic odor

Batch number 21531 FE6

Purity 100 %

Stability of test item Stable under storage conditions.

Expiry date 15-MAR-2011

Stability of test item dilution Unknown in purified water; is excluded from the statement of compliance.

Storage conditions At room temperature (range of 20 ± 5 °C), light protected.

Safety precautions Routine hygienic procedures were used to ensure the health and safety of the personnel.



## 5.4 VEHICLE

The following information was provided by RCC Ltd:

Purified water prepared at RCC Ltd (deionised water which was processed and treated by the PURELAB Option-R unit. This latter links four purification technologies: reverse osmosis, adsorption, ion-exchange and photo oxidation).

Purified water was found to be a suitable vehicle.

The vehicle was chosen after a non-GLP solubility trial which was performed before the study initiation date. This formulation trial is excluded from the GLP statement of compliance.RCC STUDY NUMBER B04803 Report Page 12

Ciba EFKA 8530



## 5.5 DOSE FORMULATION

Dose levels are in terms of the test item as supplied by the sponsor.

The dose formulations were made shortly before each dosing occasion using a magnetic stirrer as homogenizer.

The test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle added (weight:volume).

Homogeneity of the test item in the vehicl e was maintained during administration using a magnetic stirrer.



## 5.6 TREATMENT

The animals received a single dose of the te st item by oral gavage administration at 2000 mg/kg body weight after being fasted for approximately 17 hours (access to water was permitted). Food was provided again approximately 3 hours after dosing.

The dosing volume was 10 mL/kg body weight.

Rationale: Oral administration was considered to be an appropriate application method as it is a possible route of human exposure during manufacture, handling and use of the test item.



## 5.7 OBSERVATIONS

Mortality / Viability Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15.

Body weights On test days 1 (prior to administration), 8 and 15.

Clinical signs Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1. On ce daily during days 2-15. All

abnormalities were recorded.



# 6 PATHOLOGY

6.1 NECROPSY

The animal which was killed in extremis for et hical reasons on test day 13 by intraperitoneal injection of pentobarbitone at a dose of at least 2.0 mL/kg was necropsied as soon as it was

killed.

All surviving animals were killed at the end of the observation period by Carbon dioxide asphyxiation and discarded after macroscopic examinations were performed. No organs or

tissues were retained.RCC STUDY NUMBER B04803 Report Page 13

Ciba EFKA 8530



# 7 STATISTICAL ANALYSIS

No statistical analysis was used.



# 8 DATA COMPILATION

Body weights were recorded on-line.

Clinical signs were recorded on data sheets.

Mortality/viability were compiled into the RCC Tox Computer System during recording and/or

recorded on data sheets.

Macroscopic findings were compiled into the RCC Tox Computer System during recording.

The RCC Tox Computer System (RCC-Tox-Lims) had been validated with respect to data collection, storage and retrievability.RCC STUDY NUMBER B04803 Report Page 14

Ciba EFKA 8530



# 9 RESULTS

9.1 MORTALITY

One animal was killed in extremis for ethical reasons on test day 13.



## 9.2 CLINICAL SIGNS

Slightly to moderately ruffled fur was noted in all animals from the 30-minute, 1-hour and 2- hour reading to test day 2 and persisted in two animals until test day 3 and in one animal until test day 4. Hunched posture was noted in all animals of the second treated group from the 30-minute or 1-hour reading to the 5-hour reading and slight sedation was noted in the same animals from the 2- to the 5-hour reading. The animal killed in extremis for ethical reasons showed slight to moderate ruffled fur and hunched posture from test day 11 to 13, labored respiration from test day 8 to 13 and cyanosis on test day 13.



## 9.3 BODY WEIGHTS

One animal showed a slight loss of body weight (1.8 %) during the last week of observation. The body weight of the other animals was within the range commonly recorded for this strain and age.



## 9.4 MACROSCOPIC FINDINGS

Not collapsed lungs and the disgestif tract (stomach, duodenum, jejunum, ileum, caecum, colon and rectum) distended with gas was noted at the unscheduled necropsy of the animal killed in extremis for ethical reasons. No macroscopic findings were recorded at all other

necropsy.



## 9.5 MEDIAN LETHAL DOSE

The median lethal dose of Ciba EFKA 8530 after single oral administration to female rats, observed over a period of 14 days is:

LD 50 (female rat): greater than 2000 mg/kg body weightRCC STUDY NUMBER B04803 Report Page 15

Ciba EFKA 8530



# 10 INDIVIDUAL FINDINGS

10.1 MORTALITY / CLINICAL SIGNS

Dose Ani- Test days mg/kg mal Se

x Signs 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

bw No. 0.5* 1* 2* 3* 5*

2000 No clinical signs √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ 1 F

Ruffled fur 1 1 1 1

No clinical signs √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ 2 F

Ruffled fur 1 1 1 1

No clinical signs √ √ √ √ √ √ √ √ √ √ √ √ √ √ √ 3 F

Ruffled fur 1 1 1 1

2000 No clinical signs √ √ √ √ √ √ √ √ √ √ √

Ruffled fur 1 1 1 1 1 1 1

Hunched posture √ √ √ √ √ 4 F

Sedation 1 1 1

No clinical signs √ √ √ √ √ √ √ √ √ √ √ √

Ruffled fur 1 1 1 1 1 1

Hunched posture √ √ √ √ 5 F

Sedation 1 1 1

No clinical signs √ √ √ K

Ruffled fur 1 1 2 2 2 1 1 1 1 1 2

Hunched posture √ √ √ √ √ √ √ √

Sedation 1 1 1

Labored respiration √ √ √ √ √ √ 6 F

cyanosis √

Key: 1 slight, 2 moderate, 3 marked, √ noted, K killed in extremis * Examinations were performed approximately 0.5, 1, 2, 3 and 5 hours after treatment.



### No clinical signs were evident in any animal during the acclimatization period.RCC STUDY NUMBER B04803 Report Page 16

Ciba EFKA 8530



## 10.2 BODY WEIGHTS

Day 1 Day 8 Day 15 Dose mg/kg Animal

No. Se

x (treatment)

2000 1 F 190.6 189.2 185.8

2 F 190.3 203.2 215.4

3 F 196.1 215.9 226.4

2000 4 F 187.2 217.7 229.3

5 F 179.4 196.8 214.3

6 F 199.9 218.2 ---



### Body weights are presented in grams.



## 10.3 MACROSCOPIC FINDINGS

Dose mg/kg Animal

No. Se

x Mode of

death Findings

2000 1 F S No macroscopic findings

2 F S No macroscopic findings

3 F S No macroscopic findings

2000 4 F S No macroscopic findings

5 F S No macroscopic findings

6 F K Not collapsed lungs; di gestif tract (stomach, duodenum, jejunum, ileum, caecum, colon and rectum) distended with gas

S: scheduled necropsy, K: killed in extremisRCC STUDY NUMBER B04803 Report Page 17

Ciba EFKA 8530



# 11 GLP – CERTIFICATION