Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
17 September 2007 to 29 November 2007
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study performed in accordance with draft OECD test guideline in compliance with GLP and reported with a valid GLP certificate. Klimish 2 reliability assigned due to the fact that the study is still a draft guideline and not formally accepted.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 434 (Acute Dermal Toxicity - Fixed Dose Procedure)
Deviations:
yes
Remarks:
males and females where used where the guideline only recommends use of females
Principles of method if other than guideline:
OECD Guidelines for the Testing of Chemicals are periodically reviewed in the light of scientific progress and animal welfare considerations. The original acute Dermal Toxicity Guideline TG 402 was adopted in 1987. Development of a dermal Fixed Dose Procedure (FDP) was considered appropriate, following adoption of the revised Oral FDP, OECD Guideline 420 and deletion of OECD Guideline 401 in December 2001. This FDP guideline will allow the use of a series of fixed doses for the determination of acute dermal toxicity in only one sex (usually females). This study utilises both males and females. This is not expected to impact on the findings of the study.
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
not specified
Details on test material:
Identification: DUSANTOX 86Batch number: 005/07Quality control protocol No 040707/A9Storage conditions: The test article was maintained in a dark package, in the room No. 252 (at room temperature) of the laboratory of toxicology. Application form was prepared closely prior to application in laboratory 264. Long term storage: protected against sunlight. Stable: for 24 months.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Species: RatsStrain: WistarWeight: 140 – 160gNumber and sex: 3 females, 3 males – we used only 3 females and 3 males because of low acute toxicity of test article we could use limit test.Source: Accredited Breeding Velaz Prague CZIdentification: The animals were housed individually in cages after application. They were marked by number placed on the cages.Husbandry:Housing: Animals were housed in cage on bedding in groups by 5 in room No. 133 of experimental animal house. After application they were house individually. Environment: Environmental controls for the animal room will be set to maintain 22 ± 2°C, a relative humidity of 55 ± 5%. The temperature and relative humidity of air will be registered and the records maintained in animal house. A minimum of 10 air changes/air, and artificial light regime 12h light/12h dark. The sanitation was made according to standard operation procedure. Food and water: A standard certified laboratory diet (supplier Top Dovo Dobrá Voda) was served ad libitum and unlimited supply of drinking water. The diet was routinely analysed by the manufacturer for nutritional components and environmental contaminants. Acclimatisation: 5 days before the beginning of treatment.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
olive oil
Details on dermal exposure:
Approximately 24 hours before dose application, fur was removed from the dorsal area of the trunk of the test animals by clipping and shaving. The test article was applied uniformly as much of the area was covered with as thin and uniforms a film as possible on over an area which was approximately 10% of a total body surface. Test article was held in contact with the skin with a porous gauze patch covered with nonpermeabile folio by means of a semi-occlusive dressing and non-irritating tape throughout a 24-hours exposure period. At the end of the exposure period residual test article was removed, using water, without altering the existing response or integrity of the epidermis.
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
3 males and 3 females
Control animals:
no
Details on study design:
Animal dosage: On the base of literature data a dose level of 2000 mg/kg for a limit test was selected. This dose was applied to 3 females and three males by dose 2000 mg/kg. Subsequently, observations of effects and death were made. Clinical observations: Animals were observed individually after dosing at least once during the first 30 minutes, periodically during the first 24 hours, with special attention given during the first 3 hours, and daily thereafter, for a total of 14 days. All observations were systematically recorded, with individual records being maintained for each animal. Clinical signs and conditions associated with pain, suffering and impending death are described in detail in a separate OECD Guidance Document. Observations included changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems, and somatomotor activity and behaviour pattern. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. Bodyweight: Individual bodyweights of animals were determined shortly before the test article was administered and weekly thereafter. Weight changes after first and second week after application were calculated and recorded. Pathology: All test animals (including those that died during the test) were subjected to gross necropsy. All gross pathological changes were recorded for each animal. Animals to be sacrificed were anesthetised, extinguished and necropsied according to standard procedures, euthanasia and autopsy of animal

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
None of the animals died in the first 4 hours or in the 14 days observation period.
Clinical signs:
None of the animals displayed any toxic symptoms in the first 4 hours or in the 14 days observation period.
Body weight:
In first week after application 1 female did not reach weight gain. Otherwise animals reached bodyweight gains from 10 to 40 g per week.
Gross pathology:
None of animals displayed any pathological lesions.
Other findings:
Well defined or slight erythema was observed at the site of application; this subsequently healed within the study period.

Any other information on results incl. tables

Tab. 1

Sex: females     Dose: 2000 mg/kg

Animal no.

Weight [g]

Weight [g]

Weight

Weight [g]

Weight

Appl. Vol.

Response

Initial

After 1 week

Change [g]

After 2 weeks

Change [g]

ml

1

160

170

10

180

10

0.400

O

2

170

180

10

190

10

0.425

O

3

170

170

0

180

10

0.425

O

 

Tab. 2

Sex: males         Dose: 2000 mg/kg

Animal no.

Weight [g]

Weight [g]

Weight

Weight [g]

Weight

Appl. Vol.

Response

Initial

After 1 week

Change [g]

After 2 weeks

Change [g]

ml

4

220

230

10

255

25

0.550

O

5

210

250

40

275

25

0.525

O

6

240

250

10

265

15

0.600

O

 

Tab. 3

Sex: females     Dose: 2000 mg/kg

Animal no.

Signs of toxicity

Immediately

After 30 min.

After 1 hour

After 4 hours

1

w/s

w/s

w/s

w/s

2

w/s

w/s

w/s

w/s

3

w/s

w/s

w/s

w/s

 

Tab.4

Sex: males         Dose: 2000 mg/kg

Animal no.

Signs of toxicity

Immediately

After 30 min.

After 1 hour

After 4 hours

4

w/s

w/s

w/s

w/s

5

w/s

w/s

w/s

w/s

6

w/s

w/s

w/s

w/s

w/s – without symptoms

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated informationCriteria used for interpretation of results: EU
Conclusions:
The LD50 of DUSANTOX 86 after single oral application is above limit dose of 2000 mg/kg.
Executive summary:

The test article DUSANTOX 86 was applied to 3 rats females and 3 rats males in a single dermal dose 2000 mg/kg.Dose 2000 mg/kg was selected for a limit test due to presupposed low toxicity. Subsequently, observations of effects and death were made. Death and/or any toxicity symptoms did not occur.The experiment determined that the LD50 is above limit dose of 2000 mg/kg and DUSANTOX 86 fulfils conditions of category 5 “Unclassified” according to GHS. There was no observed difference in sensitivity on tested article between sexes.

No classification is applicable.