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Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 December 2013 - 05 March 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 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class 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:
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: 205 g (range: 199 g to 209 g)
- 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: 20 January 2014 to 11 February 2014

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/mL
- Justification for choice of vehicle: as unsatisfactory solubility of the test item was obtained in drinking water treated by reverse osmosis and then 0.5% methylcellulose aqueous solution (i.e. heterogeneous suspension at the concentration of 200 mg/mL was obtained), corn oil was used. A visually homogenous suspension was obtained at the concentration of 200 mg/mL in corn oil.
- Maximum dose-volume applied: 10 mL/kg

DOSAGE PREPARATION (if unusual): the test item was administered as a homogenous suspension in the vehicle.
Fresh dose formulations were prepared by the CiToxLAB France Pharmacy on the day of each administration and kept at room temperature prior to administration.

CLASS METHOD (if applicable):
- Rationale for the selection of the starting dose was selected in agreement with the Sponsor, based on the suspected low toxicity of the test item data: no morbidity or mortality was expected to occur at the dose-level of 2000 mg/kg. This was therefore chosen as the starting dose-level.
Doses:
2000 mg/kg
No. of animals per sex per dose:
6
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: the day of group allocation, just before treatment on day 1; then on days 8 and 15.
- Necropsy of survivors performed: yes (macroscopic).

Results and discussion

Effect levels
Sex:
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 were observed in any animals.
Body weight:
When compared to historical control data, a slightly lower body weight gain was noted in one female from group 1 and in two females from group 2 (+12 g or +15 g vs. +20 g ± 6.7 g in control data base) between Day 8 and Day 15. The body weight of animals was unaffected by the test item treatment.
As these differences in the mean body weight gain were minimal (and not associated to adverse effects), they were considered not to be toxicologically relevant. (see Table 1)
Gross pathology:
There were no macroscopic post-mortem observations.

Any other information on results incl. tables

Table 1

 

Sex

Female

Group

CiToxLAB France historical control data

1

2

Dose-level (mg/kg)

0

2000

2000

Body weight (mean (± SD))

 

 

 

. Day 1

198 (± 6.6)

204 (± 4.6)

206 (± 4.4)

. Day 8

239 (± 8.6)

245 (± 7.5)

248 (± 7.4)

. Day 15

258 (± 8.6)

265 (± 5.7)

264 (± 12.5)

Body weight change (mean (± SD))

 

 

 

. Days 1-8

+41(± 7.2)

+41 (± 5.2)

+42 (± 6.0)

. Days 8-15

+20 (± 6.7)

+20 (± 9.7)

+16 (± 5.1)

. Days 1-15

+60 (± 6.0)

+61 (± 9.8)

+58 (± 10.2)

SD: standard deviations.

 

Applicant's summary and conclusion

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

The potential acute toxicity of EKKE was evaluated following a single oral administration (gavage) to rats. This study was conducted in compliance with OECD Guideline No. 423 and the principles of Good Laboratory Practices. The test item, was administered once by oral route (gavage) to two groups of three fasted female Sprague-Dawley rats under a dosage-volume of 10 mL/kg. The test item was prepared incorn oil.

Based on the expected low toxicity of the test item data, the starting dose-level of 2000 mg/kg was chosen. After the first assay, as no toxicity was observed, the results were confirmed in other females. Each animal was observed at least once a day for mortality and clinical signs for 15 days. 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. No tissues were preserved.  

No unscheduled deaths occurred during the study and no clinical signs were observed in any animals. When compared to historical control data, a slightly lower body weight gain was noted in 3/6 females between Day 8 and Day 15. The body weight of the animals was unaffected by the test item treatment. As these differences in the mean body weight gain were minimal (not associated to adverse effects), they were considered not to be toxicologically relevant. At the end of the observation period, there were no macroscopic findings at necropsy. The oral LD0 of EKKE was higher than 2000 mg/kg in rats. Therefore, EKKE should not be classified for the acute toxicity by oral route according to the CLP and GHS criteria.