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 ..................................................................................................17
RCC 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) 
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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. 



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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 17th 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. 
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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 acclimatization 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 clinical 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: 

LD50 (female rat): greater than 2000 mg/kg body weight 
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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 conditions, 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 10-15 

air 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). 
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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 of 

bacteriological, 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. 
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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 vehicle was maintained during administration using a 

magnetic stirrer. 

5.6 TREATMENT 

The animals received a single dose of the test 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. Once daily during days 2-15. All 

abnormalities were recorded. 

6 PATHOLOGY 

6.1 NECROPSY 

The animal which was killed in extremis for ethical 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. 
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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. 
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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: 

LD50 (female rat): greater than 2000 mg/kg body weight 
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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 1 F No clinical signs √ √ √ √ √ √ √ √ √ √ √ √ √ √ √

Ruffled fur 1 1 1 1 

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

Ruffled fur 1 1 1 1 

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

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. 
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Ciba EFKA 8530 

10.2 BODY WEIGHTS 

Dose Day 1 Day 8 Day 15 

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; digestif tract (stomach, duodenum, 

jejunum, ileum, caecum, colon and rectum) distended with gas

S: scheduled necropsy, K: killed in extremis 
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Ciba EFKA 8530 

11 GLP – CERTIFICATION