Registration Dossier

Toxicological information

Acute Toxicity: dermal

Currently viewing:

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 December 2013 -- 14 February 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Compliant to GLP and testing guidelines; adequate consistence between data, comments and conclusions.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: breeder: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: approximately 8 weeks old on the day of treatment
- Mean body weight at study initiation for the females: 216 g and 348 g for the males .
- Fasting period before study: yes, during the night before treatment
- Housing: polycarbonate cages
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: at least 5 days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h

IN-LIFE DATES: 27 January 2014 to 14 February 2014

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: 10% of body surface, dorsal site
- Type of wrap if used: hydrophilic gauze pad + adhesive hypoallergenic aerated semi-occlusive dressing + restraining bandage

REMOVAL OF TEST SUBSTANCE
- Removal of dressing: 24h post-exposure
- Washing: at 24h post-exposure, with a moistened cotton pad with water for injection

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg
- Constant volume: no
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Clinical observations: frequently during the hours following treatment; then, at least once a day.
- Body weight: just before treatment, on Day 1; then on Days 8 and 15.
- Necropsy of survivors performed: yes (macroscopic).

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD0
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no mortality
Mortality:
No unscheduled deaths occurred during the study.
Clinical signs:
No clinical signs indicative of systemic toxicity were observed in any animals.
A very slight to well-defined erythema was noted in 4/5 females between Day 3 and Day 9 and in 4/5 males between Day 3 and Day 5. Scabs were observed in 2/5 females from Day 7 until Day 11 or 14 and in 1/5 males from Day 13. Dryness of the skin was noted in 2/5 females from Day 7.
Body weight:
The mean body weights and the mean body weight changes (g) recorded in test item-treated animals during the observation period and in historical control data are summarized in the Table 1.
When compared to CiToxLAB France historical control data, a lower body weight gain was noted in 1/5 females (+3 g vs. +27 g ± 11.1 g in control data base) between Day 8 and Day 15 and in 3/5 females (+32 g or +35 g vs. +52 g ± 12.9 g in control data base) between Day 1 and Day 15. A lower body weight gain was also noted in 4/5 males (+27 g to +33 g vs. +50 g ± 12.0 g in control data base) from Day 8 to Day 15 and over the whole study period (+60 g to +69 g vs. +89 g ± 12.8 g in control data base).
Nevertheless, compared to CiToxLAB France historical control data, the body weight was unaffected by the test treatment in both sexes.
Gross pathology:
A 0.8 cm-long x 0.2 cm-wide scab was recorded in one male at the application site. Although this finding was not seen in any other animals in the study, a relationship to EKKE administration could not be excluded.

Any other information on results incl. tables

Table 1

 

Sex

Female

Male

Group

CiToxLAB France Historical control data

1

CiToxLAB

France Historical control data

2

Dose-level (mg/kg)

0

2000

0

2000

Body weight (mean (± SD))

 

 

 

 

. Day 1

217 (± 10.3)

216 (± 6.9)

333 (± 10.3)

348 (± 22.4)

. Day 8

242 (± 10.6)

237 (± 8.9)

372 (± 9.5)

383 (± 23.1)

. Day 15

269 (± 13.0)

256 (± 14.4)

422 (± 12.6)

416 (± 25.9)

Body weight change (mean (± SD))

 

 

 

 

. Days 1-8

+25 (± 10.0)

+21(± 4.9)

+39 (± 11.5)

+35 (± 4.0)

. Days 8-15

+27 (± 11.1)

+19 (± 11.6)

+50 (± 12.0)

+34 (± 6.3)

. Days 1-15

+52 (± 12.9)

+40 (± 7.9)

+89 (± 12.8)

+68 (± 8.2)

SD: standard deviations.

 

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions of this study, the dermal LD0 of the test item, was higher than 2000 mg/kg in rats.
Therefore, EKKE should not be classified for the acute dermal toxicity route according to the CLP and GHS criteria.
Executive summary:

The potential toxicity of EKKE was evaluated following a single dermal application to rats. This study was conducted in compliance with OECD Guideline No. 402 and the principles of Good Laboratory Practices. The test item, was applied in its original form to the skin of five female then five male Sprague-Dawley rats at the dose-level of 2000 mg/kg. The application site was covered by a semi-occlusive dressing for 24 hours. Each animal was observed at least once a day for mortality and clinical signs for 15 days. From Day 2, any local reactions at the treatment site were also noted. Body weight was recorded on Day 1 and then on Days 8 and 15. On completion of the observation period, the animals were sacrificed and then submitted for a macroscopic post-mortem examination. Macroscopic lesions were preserved in buffered formalin then destroyed at the finalization of the study report as no microscopic examination was performed.  

No unscheduled deaths and no clinical signs indicative of systemic toxicity were observed in any animals. A very slight to well-defined erythema was noted in 4/5 females between Day 3 and Day 9 and in 4/5 males between Day 3 and Day 5. Scabs were observed in 2/5 females from Day 7 until Day 11 or 14 and in 1/5 males from Day 13. Dryness of the skin was noted in 2/5 females from Day 7. When compared to historical control data, a lower body weight gain was noted in 1/5 females (+3 g vs.+27 g ± 11.1 g in control data base) between Day 8 and Day 15 and in 3/5 females (+32 g to +35 g vs.+52 g ± 12.9 g in control data base) between Day 1 and Day 15. A lower body weigh gain was also noted in 4/5 males (+27 g to +33 g vs.+50 g ± 12.0 g in control data base) from Day 8 to Day 15 and over the whole study period (+60 g to +69 g vs.+89 g ± 12.8 g in control data base).

Nevertheless, compared to CiToxLAB France historical control data, the body weight was unaffected by the test treatment in both sexes. There were no test item-related macroscopic post-mortem findings, with the exception of equivocal scab in one male.  

The dermal LD0 of EKKE was higher than 2000 mg/kg in rats. Therefore, the test item should not be classified for the acute dermal toxicity according to the CLP and GHS criteria.