Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28-01-2014 to 04-03-2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met.
Justification for type of information:
Information as to the availability of the in vivo study is provided in 'attached justification'.

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
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries, Test Data for Registration of Agricultural Chemicals, Acute oral toxicity (2-1-1), 12 Nousan No 8147, Agricultural Production Bureau, November 24, 2000
GLP compliance:
yes (incl. certificate)
Remarks:
inspected: November 2012; signature: April 2014
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Physical state: Liquid
- Storage condition of test material: ca. 4°C in the dark under nitrogen
- Other: colourless

Test animals

Species:
rat
Strain:
other: Crl:CD (SD)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Recognised supplier
- Females (if applicable) nulliparous and non-pregnant: Yes.
- Age at study initiation: 8 - 12 weeks.
- Weight at study initiation: 198 - 239 g
- Fasting period before study: Overnight before dosing and approximately four hours after dosing.
- Housing: Group housed in groups of up to three same sex in suspended solid-floor polycarbonate cages furnished with softwood flakes and aspen chew block as cage enrichment..
- Diet (e.g. ad libitum): Certified diet from recognised supplier, provided ad libitum (except for fasting period).
- Water (e.g. ad libitum): ad libitum (except for fasting period)
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 23
- Humidity (%): 40 - 70%
- Air changes (per hr): Not reported. Positive pressure, filtered air environment.
- Photoperiod: 12 h light / 12 h dark

IN-LIFE DATES: From: To: 2014-01-28 to 2014-03-04

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: The test item was formulated at concentrations of 30 or 200 mg/mL in the vehicle and administered at a volume of 10 mL/kg body weight. Formulations were prepared on the day of dosing.
- Amount of vehicle (if gavage): The concentration of the test item in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed.
- Lot/batch no. (if required): See full study report.
- Purity: See full study report.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw.

DOSAGE PREPARATION (if unusual): Not applicable. The test item was prepared in the vehicle. It was administered to the animals under a volume of 10 mL/kg. The volume administer was adjusted according to bodyweight on day of treatment.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: 300 mg/kg bw based on guideline recommendations. In the absence of data regarding the toxicity of the test item, 300 mg/kg was chosen as the starting dose.
Doses:
300 mg/kg bw in corn oil vehicle (starting dose); 2000 mg/kg bw
No. of animals per sex per dose:
3 initial females (sighting study) and further 3 females (main study); total of up to 6 per dose according to sequential guideline testing strategy.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made 0.5, 1, 2, and 4 hours after dosing and then daily for fourteen days. Morbidity and mortality checks were made twice daily. Individual body weights were recorded on Day 1 (the day of dosing) and on Days 8 and 15.
- Necropsy of survivors performed: yes

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
300 mg/kg bw: No mortality
2000 mg/kg bw: Two females mortalites on day 2 (B12) and 7 (B10) respectively.
Clinical signs:
300 mg/kg bw: No signs of systemic toxicity were noted.
2000 mg/kg bw: Clinical signs prior to death comprised of piloerection and elevated gait (both B10 and B12), hunched posture (B10) and loose faeces (B12). Surviving female signs comprised salivation, chin rubbing, under activity, piloerection, elevated gait and loose faeces in one female (B9). Signs were first noted approximately five minutes after dosing. Recovery was complete by Day 2. No signs were observed in other survivors.
Body weight:
300 mg/kg bw: A slight bodyweight loss or low body weight gain was noted between Days 8 and 15 for several females receiving 300 mg/kg/day. All other females were considered to have achieved satisfactory body weight gains. As no such effects were seen at 2000 mg/kg/day the minor bodyweight loss in females receiving 300 mg/kg/day was not considered treatment related.
2000 mg/kg bw: Survivors demonstrated bodyweight gains over the study period. The largest increase was demonstrated by female B11 (+38 grams and +13 grams) in days 1-8 and 8-15 respectively.
Gross pathology:
300 mg/kg bw: No abnormalities were noted at necropsy.
2000 mg/kg bw: Macroscopic examination at study termination on Day 15 revealed pallor of the liver and kidneys in one female (B11). No abnormalities were noted at necropsy amongst other survivors.
In the non-survivors: congestion (characterised by darkened tissues/organs) of the subcutaneous tissue, lungs, liver, spleen and kidneys, clear fluid content in the thoracic cavity, pallor of the stomach, small caecum, yellow fluid content in the small intestine and duodenum (both B11 and B12). Enlarged spleen, red fluid content in the duodenum and gaseous distension in the small and large intestine (B10) and yellow fluid content in the stomach, gaseous distension in the duodenum and yellow fluid content in the large intestine (B12)
Other findings:
- Organ weights: Not reported.
- Histopathology: Not reported. No macropathological abnormalities (coupled with relevant clincial signs) amongst survivors.
- Potential target organs: Not applicable. No macropathological abnormalities (coupled with relevant clincial signs) amongst survivors.
- Other observations: Not applicable.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study the oral LD50 was established to be > 300 and < 2000 mg/kg bw in female Crl:CD (SD) rats.
Executive summary:

The study was performed according to OECD TG 423, EU Method B.1 tris, US EPA OPPTS 870.1100 and Japan Acute Oral Toxicity (2-1-1) (2000) guidelines in accordance with GLP to assess the acute oral toxicity of the test item by the acute toxic class method following a single oral administration in the female Crl:CD (SD) strain rat. The test item was administered by oral gavage in corn oil vehicle at a dose volume of approximately 10 mL/kg in an initial step at 300 mg/kg in three females. In the absence of significant toxicity, at this dose level the test item was then administered again in a further three females. In the absence of toxicity the test item was finally administered at 2000 mg/kg bw in corn oil by oral gavage to a group of three females. At 300 mg/kg bw there was no mortality, no significant clinical signs and a slight bodyweight loss or low body weight gain was noted between Days 8 and 15 for most females receiving 300 mg/kg/day. As no such effects were seen at 2000 mg/kg/day the minor bodyweight loss in females receiving 300 mg/kg/day was not considered to be related to the treatment. There was no abnormalities at necropsy. At 2000 mg/kg bw, there was two mortalities on day 2 and 7. Clinical signs prior to death comprised of piloerection and elevated gait (both animals), and then loose faeces and hunched posture, respectively. Clinical signs for survivors included salivation, chin rubbing, under activity, piloerection, elevated gait and loose faeces in one female. Recovery, as judged by external appearance and behaviour, was complete by Day 2. All survivors gained bodyweight. Macroscopic examination at study termination on Day 15 revealed pallor of the liver and kidneys in one female. Under the conditions of this study the oral LD50 was established to be > 300 and < 2000 mg/kg bw in the female rat.