Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-03-08 to 2017-05-29
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 0016044582

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Young adult animals (female animals approx. 10 weeks)
- Weight at study initiation: Animals of comparable weight (± 20% of the mean weight, actual weights see Appendix I)
- Fasting period before study: at least 16 hours before administration
- Housing: Single housing
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 +/- 3 °C
- Humidity: 30 – 70 %
- Air changes per hr: approx. 10
- Photoperiod: 12 / 12 hrs dark / hrs light

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 20 g/mL
- Amount of vehicle: 10 mL/kg bw
- Justification for choice of vehicle:
A homogeneous mixture of the test item in water could not be guaranteed because the test item preparation was a suspension. Therefore, a 0.5% solution of CMC in deionized water was used as vehicle.

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg bw

DOSAGE PREPARATION:
The test item was ground with a mortar and pestle. Thereafter, the test item preparation for each test group was produced shortly before administration by stirring with a high speed homogenizer (Ultra-Turrax) and a magnetic stirrer. The homogeneity of the test item preparation during administration was ensured by stirring with a magnetic stirrer.


CLASS METHOD
- Rationale for the selection of the starting dose: based on available data, the substances was not considered to be very toxic
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
2 x 3 females per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Individual body weights shortly before administration (day 0), several times thereafter in some animals, in general weekly and on the last day of observation, or on day of death on study day 2 in one animal. Clinical signs for each animal were recorded several times on the day of administration and at least once during each workday thereafter. A check for any dead or moribund animals was made at least once each workday; these records are archived by Bioassay.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Statistics:
Calculations were performed using Microsoft Excel 2010 and checked with a calculator.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred in the first 2000 mg/kg bw test group, while 1 animal in the second 2000 mg/kg bw test group died at study day 2 after administration.
Clinical signs:
In the first 2000 mg/kg bw test group impaired general state and piloerection were noted in all animals from hour 1 until study day 3, 6 or 10 after administration, while dyspnoea was observed in these animals from hour 1 until study day 2. Cowering position was noticed on study day 1 in all animals and persisted in two animals up to study day 2. Furthermore, reduced defecation and diarrhea was seen in all animals on study day 1; reduced defecation was noted again in one of these animals on study day 6. Non defecation was observed in one animal on study day 2 while another animal showed this finding from study day 2 until study day 3. In addition exsiccosis was observed in all animals on study day 1 and persisted in two animals until study day 3.

Chromodacryorrhea was noticed in one animal on study day 1 while in another animal smeared fur with feces was observed on the same day.
In all animal loss of body weight was seen during the first 2 or 3 days after administration.

In both surviving animals of the second test group impaired general state and piloerection were observed from hour 1 until study day 3, while dyspnoea was noticed in these animals from hour 1 until study day 1. Furthermore, exsiccosis and diarrhea was seen in these animals on study day 1, while reduced defecation was seen from study day 1 until study day 2. In the animal that died in this test group impaired general state was observed from hour 1 until hour 5, while poor general state was noticed on study day 1. In addition, piloerection and dyspnoea were seen in this animal from hour 1 until study day 1, while cowering position was noticed from hour 2 until study day 1. From hour 2 until hour 3 and on study day 1, tremor was seen, while twitching was noticed from hour 4 until hour 5 in this animal. Additionally, diarrhea, reduced defecation and exsiccosis were observed in this animal on study day 1.
Body weight:
The body weight of the animals in the first 2000 mg/kg bw test group decreased within the first 2 or 3 days after administration. Thereafter, the body weight slightly increased until the end of the first observation week. In the second week, the body weight increased within the normal range in these animals.
The body weights of the surviving animals in the second 2000 mg/kg bw test group increased within the normal range throughout the study period.
Gross pathology:
Due to the advanced putrefaction no macroscopic pathological findings could be determined in the animal that died in the second 2000 mg/kg bw test group.
There were no macroscopic pathological findings in the surviving animals sacrificed at the end of the observation period (5 females).

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met