# RCC Study Number 848622

DEA/Acid anhydride reaction product: Acute Dermal Toxicity Study in Rats
Report



## Author: G. Arcelin Sponsor: Rockwoollnternational A/S Hovedgaden 584 DK-2640 Hede;husene 

## I Denmark

Page 1 of 17RCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product 

## Table of contents Report Page2



### 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 UNIT................................................................................6 1.7 STATEMENT OF COMPLIANCE ...........................................................................7 1.8 TEST GUIDELINES ...............................................................................................8 1.9 ANIMAL WELFARE................................................................................................8 2 SUMMARY OF RESULTS ...............................................................................................9 3 CONCLUSION ................................................................................................................9 4 PURPOSE .....................................................................................................................10 5 MATERIALS AND METHODS .......................................................................................10 5.1 TEST SYSTEM ....................................................................................................1 0 5.2 HUSBANDRY.......................................................................................................10 5.3 TEST ITEM ..........................................................................................................11 5.4 TEST ITEM PREPARATION ................................................................................11 5.5 TREATMENT .......................................................................................................12 5.6 OBSERVATIONS .................................................................................................12 6 PATHOLOGY ................................................................................................................12 6.1 NECROPSY .........................................................................................................12 7 STATISTICAL ANALYSIS .............................................................................................13 8 DATACOMPILATION ...................................................................................................13 9 RESULTS ......................................................................................................................14 9.1 MORTALITY.........................................

**.-~** 

### .............................................................. 14 9.2 CLINICAL 

### I LOCAL SIGNS

..................

**~~** 

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

**~** 

### .................... 14 9.4 MACROSCOPIC FINDINGS ................................................................................14 9.5 MEDIAN LETHAL DOSE .......................................................................................14 10 INDIVIDUAL FINDINGS ................................................................................................15 10.1 CLINICAL SIGNS .................................................................................................15 10.2 BODY WEIGHTS .................................................................................................16 10.3 MACROSCOPIC FINDINGS ................................................................................16 11 GLP- CERTIFICATION ................................................................................................17RCC STUDY NUMBER 848622 DEAl Acid anhydride reaction product Report


## 1 PREFACE 1.1 GENERAL Title DEAl Acid anhydride reaction product: Acute Dermal Toxicity Study in Rats Sponsor Rockwoollnternational A/S Hovedgaden 584 Study Monitor

Test Facility DK-2640 Hedehusene 

### I Denmark Mr. Flemming A. Simonsen, DVM Danish Toxicology Centre 2 Kogle Aile DK-2970 Hoersholm 

### I Denmark RCC Ltd Toxicology 

# w olferstrasse 4 CH-4414 FOIIinsdorf 

### I Switzerland 

## 1.2 RESPONSIBILITIES Study Director Deputy for Study Director Technical Coordinator Head of RCC Quality Assurance 

## 1.3 SCHEDULE G. Arcelin M. Ott N. Hostettler 

### 1. Wuthrich

Experimental Starting Date 13-MA Y-2003 Experimental Completion Date 03-JUN-2003 Delivery of Animals 13-MA Y-2003 Acclimatization Treatment Observation Study Completion Date 13-MA Y-2003 to 19-MA Y-2003 20-MA Y-2003 13-MAY-2003 to 03-JUN-2003 17-SEP-2003 Page3RCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product 

## 1.4 ARCHIVING Report Page4



### RCC Ltd (CH-4452 ltingen 

### I 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 848622 DEA/Acid anhydride reaction product 

## 1.5 SIGNATURE PAGE Study Director:

Management: Report PageS



# G.~ 

# /Q

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# ~~~ date:



# (f~·i-::iJ~

**~~** Dr. H. Fankhauser



# £~~/~t?JRCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product Report


## 1.6 QUALITY ASSURANCE UNIT
RCC Ltd, Toxicology, CH-4452 ltingen 

### I Switzerland
STATEMENT
RCC STUDY NUMBER : TEST ITEM STUDY DIRECTOR TITLE 848622 DEA/Acid anhydride reaction product G. Arcelin DEAl Acid anhydride reaction product: Acute Dermal Toxicity Study in Rats Page6

The general facilities and activities are inspected periodically and the results are reported to the responsible person and the management. Study procedures were periodically inspected. The study plan and this report were audited by the Quality Assurance Unit. The dates are given below.
Dates and Types of Dates of Reports to the Study Director

QAU Inspections and to Management 08-MA Y-2003 Study Plan 08-MA Y-2003 20-MAY-2003 Test System, Test item, 20-MA Y-2003 Treatment, Raw Data 25-JUN-2003 Report 25-JUN-2003

This statement also confirms that this final report reflects the raw data. Quality Assurance:

date:

# A1-St:P-20o3RCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product Report

GOOD LABORATORY PRACTICE 

## 1.7 STATEMENT OF COMPLIANCE
RCC STUDY NUMBER : TEST ITEM STUDY DIRECTOR TITLE 848622 DEAl Acid anhydride reaction product G. Arcelin DEAl Acid anhydride reaction product: Acute Dermal Toxicity Study in Rats Page?

This study has been performed in compliance with the Swiss Ordinance relating to Good Laboratory Practice, adopted February 2nd, 2000 [RS 813.016.5]. This Ordinance is based on the OECD Principles of Good Laboratory Practice, as revised in 1997 and adopted November 26th, 1997 by decision of the OECD Council [C(97) 186/Final]. Study Director:RCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product 

## 1.8 TEST GUIDELINES Report PageS

The study procedures described in this report meet or exceed the requirements of the following guidelines: OECD Guidelines for Testing of Chemicals, Section 4, Number 402

..Acute Dermal Toxicity

.. , adopted February 24, 1987. · Directive 92/69/EEC, 8.3

...Acute Toxicity-Dermal

.. , July 31, 1992. 

## 1.9 ANIMAL WELFARE This study was performed in an AAALAC-approved laboratory in accordance with the Swiss Animal Protection Law under license no. 32.RCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product Report


## 2 SUMMARY OF RESULTS Page9



### Five male and five female HanBrl: WIST (SPF) rats were treated with DEA/Acid anhydride reaction product at 2000 mg/kg by dermal application. The test item was applied undiluted as delivered by the Sponsor. The application period was 24 hours. 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 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Mortality/viability was recorded twice daily during test days 1-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically. No deaths occurred during the study. No clinical signs were observed during the observation period. Three female animals showed a loss of body weight {1.5 

### °/o to 4.7 %) 

### during the first observation week and recovered from test day 8 to the end of the observation period. The body weight of the other animals was within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy. 

## 3 CONCLUSION The median lethal dose of DEAl Acid anhydride reaction product after single dermal administration to rats of both sexes, observed over a period of 14 days is: LDso (rat): greater than 2000 mg/kg body weightRCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product 

## 4 PURPOSE Report Page 10



### The purpose of this study was to assess the acute dermal toxicity of DEA/AC"id anhydride reaction product when administered to rats by a single semi-occlusive dermai application, followed by an observation period of 14 days. This study should provide a rational basis for risk assessment. 

## 5 MATERIALS AND METHODS 5.1 TEST SYSTEM Test system Rat, HanBrl: WIST (SPF) Rationale Recognized by the international guidelines as a recommended test system. Source RCC Ltd, Laboratory Animal Services CH-4414 Fullinsdorf 

### I Switzerland Number of animals per group 5 males and 5 females Total number of animals 5 males and 5 females Age when treated Males: 8 - 9 weeks Females: 11
- 12 weeks Identification By unique cage number and corresponding color-coded spots on the tail. The animals were marked immediately prior to treatment. 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. Conditions 104 

### I RCC Ltd, FOJiinsdorf Standard Laboratory Conditions. Air-conditioned with 1 0-15 air changes per hour, and continuously monitored environment with target ranges for room temperature 22 ± 

### 3 

# oc and for relative humidity between 30-70 

### % (values above 70 

### % during cleaning process possible), 12 hours fluorescent light/ 12 hours dark, music during the light period.RCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product Report Page 12


## 5.5 TREATMENT One day before treatment, the backs of the animals were clipped with an electric clipper, exposing an area of approximately 1 0 

### °/o of the total body surface. Only those animals without injury or irritation on the skin were used in the test. On test day 1, the test item was applied at a dose of 2000 mg/kg body weight on a patch. The patch with the test item was applied on the intact skin and covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage. Application amount/kg body weight: 2 g Twenty-four hours after the application the dressing was removed and the skin was flushed with lukewarm tap water and dried with disposable paper towels. Thereafter, the reaction sites were assessed. Rationale Dermal administration was used as this is one possible route of human exposure during manufacture, handling and use of the test item. 

## 5.6 OBSERVATIONS Mortality 

### I Viability Body weights Clinical signs



# 6 PATHOLOGY 6.1 NECROPSY Daily during acclimatization and twice daily during days 1-15. On test days 1 (prior to administration), 8 and 15. Daily during acclimatization and at approximately 1, 2, 3 and 5 hours after administration on test day 1 . Once daily during days 2-15.

All animals were killed at the end of the observation period by an intraperitoneal injection of Vetanarcol at a dose of at least 2.0 mUkg body weight (equivalent to at least 324 mg sodium pentobarbitone/kg body weight) and discarded after macroscopic examinations were performed. No organs or tissues were retained.RCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product Report


## 7 STATISTICAL ANALYSIS No statistical analysis was used. 

## 8 DATA COMPILATION Body weights were recorded on-line. Clinical signs were recorded on data sheets. Page 13

Mortality/viability were compiled into the RCC computer system during recording and/or recorded on data sheets. Macroscopic findings were compiled into the RCC computer system during recording.RCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product 

## 9 RESULTS 9.1 MORTALITY No deaths occurred during the study. Report



## 9.2 CLINICAL 

## I LOCAL SIGNS No systemic or local signs of toxicity were observed during the study period. 

## 9.3 BODY WEIGHTS Page 14



### Three female animals showed a loss of body weight (1.5 

### °/o to 4.7 

### o/o) during the first observation week and recovered from test day 8 to the end of the observation period. The body weight of the other animals was within the range commonly recorded for this strain and age. 

## 9.4 MACROSCOPIC FINDINGS No macroscopic findings were observed at necropsy. 

## 9.5 MEDIAN LETHAL DOSE The median lethal dose of DEAl Acid anhydride reaction product after single dermal administration to rats of both sexes, observed over a period of 14 days is: LDso (rat): greater than 2000 mg/kg body weightRCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product Report


## 10 INDIVIDUAL FINDINGS 10.1 CLINICAL SIGNS Sex/ Animal Signs Test Day Dose No. 1 2 1* 2* 3* 5* male/ 1 No clinical signs 

# """"" 2000 

# "

# """" mg/kg 2 No clinical signs 3 No clinical signs 

# """"" 4 No clinical signs 

# """"" 5 No clinical signs 

# """"" female 6 No clinical signs 

# "

# ""

# "" /2000 

# "

# ""

# "" mg/kg 7 No clinical signs 8 No clinical signs 

# ""

# """ 9 No clinical signs 

# """"" 10 No clinical signs 

# "

# "

# """ Key: " noted 3 4 5 6 7 8 9 

# " """""" """"""" """ ""

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# """" " """""" """"""

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* Examinations were performed approximately 1, 2, 3 and 5 hours after treatment. 10 

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No clinical signs were evident in any animal during the acclimatization period. Page 15

11 12 13 14 15 

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## 10.2 BODY WEIGHTS Body weight in grams: Sex/ Dose Animal No. male I 2000 mg/kg 1 2 3 4 5 female I 2000 mg/kg 6 7 8 9 10 Report

Day of Treatment 246.0 251.6 257.3 247.9 250.1 187.0 191.6 189.4 196.4 190.1 

## 10.3 MACROSCOPIC FINDINGS

Sex/ Dose Animal No. Mode of Death male I 2000 mg/kg 1 Scheduled necropsy 2 Scheduled necropsy 3 Scheduled necropsy 4 Scheduled necropsy 5 Scheduled necropsy female I 2000 mg/kg 6 Scheduled necropsy 7 Scheduled necropsy 8 Scheduled necr9psy 9 Scheduled necropsy 10 Scheduled necropsy Page 16

DayS Day 15 267.7 285.4 266.0 281.2 278.8 312.2 265.3 283.2 260.9 278.5 195.9 205.9 194.4 195.7 186.3 201.3 187.1 194.2 187.2 196.3

Findings No macroscopic findings. No macroscopic findings. No macroscopic findings. No macroscopic findings. No macroscopic findings. No macroscopic findings. No macroscopic findings. No macroscopic findings. No macroscopic findings. No macroscopic findings.RCC STUDY NUMBER 848622 DEA/Acid anhydride reaction product Report


## 11 GLP- CERTIFICATION

The Swiss GLP Monitoring Authorities

SWiss Federal Office of Public Health Swiss Agency for the .J·. Swlssmedic Environment, Forests r . Swiss Agencyfor andlandscape SWlSSmedlC TherapeulicProducts 

# Statement of GLP Compliance

It is hereby confirmed that during the period of the following Test Facilities of November 18-22,2002 RCC Ltd 4452 ltingen Switzerland

were inspected by the Federal Office of Public Health, the Swiss Agency for Therapeutic Products and the Swiss Agency for the Environment, Forests and Landscape with respect to the compliance with the Swiss legislation on Good Laboratory Practice. Test Facilities -Toxicology

- Environmental Chemistry and

Pharmanalytics Areas ofexpertise *
TOX, ACC, MUT, OTH (Safety Pharmacology) ACC, ECT, ENF, EMN, PCT, RES, OTH (Animal metabolism)

The inspections were performed in agreement with the OECD Guidelines for National GLP Inspections and Audits. It was found that the aforementioned test facilities were operating in compliance with the Swiss Ordinance relating to Good Laboratory Practice [RS 813.016.5] at the time they were inspected.

Bern, March 2003 Federal Office of Public Health The'Director

Prof. Th. Zeltner

• TOX"' Toxicology; ACC • Analytical and Clinical Chemistry; ECT= Environmental toxicity on aquatic and terrestrial organisms ; ENF =Behaviour in water, soil and alr. Bloaccumutatfon ; EMN :a: Studies on effects on mesocosms and natural ecosystems; MUT = Mutagenicity; PCT= Physical-chemical testing ; RES= Residue studies : OTH =

**Oth~r.** to be specified. Page 17