Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08-09-2011 to 07-10-2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report Date:
2000

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)
Remarks:
inspected: September 1999 ; signature: December 1999
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: At room temperature, protected from light and under nitrogen gas
- Other: colourless

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
ICO: OFA-SD (lOPS Caw)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Recognised animal supplier
- Age at study initiation: ca. 6 weeks (nulliparous and non-pregnant)
- Weight at study initiation: females: 170 ±7 g and males: 141 ±2 g
- Fasting period before study: Overnight (ca. 18 hours) and fasting ceased ca. 4 hours after administration of the test item.
- Housing: Group housing 3 same sex animals per polycarbonate cages ( 48 cm x 27 cm x 20 cm) containing sterilized sawdust as bedding material as cage-enrichment.
- Diet: certified rat diet ad libitum
- Water: filtered certified water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ±2
- Humidity (%): 30 - 70
- Air changes (per hr): ca. 12 cycles/hr
- Photoperiod: 12 hours light / 12 hours dark

IN-LIFE DATES: From: 11-04-2000 To: 10-05-2000

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Identity: Corn Oil (batch number: reported in the full study report)
- Concentration in vehicle: The concentration of the test item in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed.

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: In the absence of data suggesting the test material was toxic, 200 mg/kg was chosen as the starting dose. The toxicity of the test item was assessed by stepwise treatment of groups of 3 males (initially) then 3 females, as appropriate. The first group was treated at a dose level of 200 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step 2000 mg/kg bw. Then a further test was conducted on three females at the highest dose level. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups..
Doses:
Initial: 200 mg/lg and final: 2000 mg/kg
No. of animals per sex per dose:
3 at the highest employed dose (3 males and 3 females)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality/Viability: Clinical signs and mortality: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The signs were graded according to fixed scales and the time of onset, degree and duration were recorded ; Bodyweights: Days 1 (pre-administration), 8 and 15
- Necropsy of survivors performed: yes

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 - < 5 000 mg/kg bw
Based on:
test mat.
Mortality:
2000 mg/kg bw: there was 1/3 female mortalities and 0/3 male mortalities. Mortality occurred on day 1 within 30 minutes of treatment.
200 mg/kg bw: there was no male mortalities.
Clinical signs:
2000 mg/kg bw: In the female mortality which occurred within 30 minutes of treatment, no clinical signs were observed. Females gave no clinical signs within the observation period. In males, no mortality occurred. Hypoactivity, piloerection, stiff gait and dyspnoea were noted in all males for 24 and 48 hours. All signs had reversed by day 3.
300 mg/kg bw: No significant clinical signs.
Body weight:
All survivors showed bodyweight gains over the study period. The body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain and consistent with laboratory controls.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination.
Other findings:
- Organ weights: Not reported.
- Histopathology: Not reported. No macropathological abnormalities.
- Potential target organs: Not applicable.
- Other observations: Not applicable.

Any other information on results incl. tables

Applicant assessment indicates: the single female mortality at 2000 mg/kg bw/day may not be test item administration related, as there is no corresponding clinical signs and abnormal macro pathological findings in other females at the same dose level. All females gained body weight during the study. Additionally, the abnormal male clinical signs were not significant and were transient - ceasing within 72 hours. Expert judgement may associate these signs with the treatment procedure. It is considered that the single female mortality is not conclusively treatment related. This is given the absence of corresponding findings and additionally given the proximity of mortality to the oral gavage treatment procedure. The LD50 cut off value of 2500 mg/kg bw has been assigned. This may increased if additional evidence becomes available conclusively indicating the single female mortality is not test item related, on the weight of evidence.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study the oral LD50 established to be > 2000 and < 5000 mg/kg bw in female Wistar rats.
Executive summary:

The study was performed according to OECD TG 423 and EU Method B1 tris guidelines in accordance with GLP to assess the acute oral toxicity of the test item following a single oral administration in the female Sprague-Dawley ICO: OFA-SD (lOPS Caw) strain male/female rat by the acute class method. The test item was administered by oral gavage in a corn oil vehicle to two sequential groups of three male rats at 200 mg/kg bw and/or three male and then three females at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15). There was no mortality, clinical signs or abnormalities in necropsy and all animals gained weight at 200 mg/kg bw. There was a single female mortality in the second group at 2000 mg/kg bw within 30 minutes of treatment. No clinical signs was observed in females. In males no mortality occurred. Hypoactivity, piloerection, stiff gait and dyspnoea were clinical signs noted in all males on days 1 and/or day 2. Recovery was complete on day 3. The body weight gain shown by survivors over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain. No abnormalities were found at macroscopic post mortem examination of the animals at any dose level. The oral LD50 value of the test item in Sprague-Dawley rats was established to be > 2000 and < 5000 mg/kg body weight.