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Toxicological information

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:
18 July 2016 to 11 August 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

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, including most recent amendments
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
Qualifier:
according to
Guideline:
other: JMAFF 12 Nousan, Notification No 8147
Version / remarks:
2000, including the most recent partial revisions
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Appearance: Yellow powder
- Storage conditions of test material: At room temperature protected from light
- Test material handling: Use amber glassware or wrap container in aluminium foil
- Stable at higher temperatures: Yes, maximum temperature: 95 °C

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: Crl:WI (Han) (outbred, SPF-Quality)
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: Young adult animals (approximately 8 - 9 weeks old)
- Weight at study initiation: 148 to 188 g (mean 163 g). Body weight variation did not exceed ± 20 % of the sex mean.
- Fasting period before study: Yes. Animals were deprived of food overnight prior to dosing and until 3 to 4 hours after administration of the test material. Water was available ad libitum.
- Housing: Group housing of 3 animals per cage in labelled cages (height 18 cm) containing sterilised sawdust as bedding material and paper as cage-enrichment
- Diet: ad libitum pelleted rodent diet
- Water: ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 18 to 24 °C
- Humidity: 40 to 70 % (relative)
- Air changes: At least 10 air changes/hour
- Photoperiod: 12-hour light/12-hour dark cycle

IN-LIFE DATES: Not reported

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: The concentration of the test material 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 preparations performed at the testing facility and on test material data supplied by the sponsor.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

DOSAGE PREPARATION: The preparations (w/w) were kept at room temperature protected from light and were dosed within 4 hours after adding the vehicle to the test material. Homogeneity was assessed by visual inspection of the solutions and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies. No correction was made for purity of the test material.

CLASS METHOD (if applicable)
- Dosing pattern: The toxicity of the test material was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. 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:
2000 and 300 mg/kg body weight
No. of animals per sex per dose:
3 animals per sex per dose group (1 and 2 groups for the 2000 and 300 mg/kg dose levels, respectively)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: Animals were observed for mortality/viability twice daily. Body weights were recorded on Days 1 (pre-administration), 8 and 15 and at death (if found dead after Day1). Clinical signs were observed 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.
- Necropsy of survivors performed: Yes. At the end of the observation period all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.
Statistics:
No statistical analysis was performed (the method used is not intended to allow the calculation of a precise LD50 value).

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
At 2000 mg/kg, two animals were found dead on Day 1 and one animal was found dead on Day 2. At 300 mg/kg, no mortality occurred.
Clinical signs:
At 2000 mg/kg, lethargy, hunched posture, rales, shallow respiration, salivation, ptosis and/or hypothermia were noted for all animals on Day 1. At 300 mg/kg, hunched posture and/or salivation were noted for all animals on Day 1. Yellow discoloration of the back, faeces, urine, forelegs, snout, paws, genital region and/or mouth was noted for most animals. This was considered due to the colour of the test material.
Body weight:
The mean body weight gain shown by the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.
Gross pathology:
At 2000 mg/kg, yellow discoloration of the contents of the stomach, duodenum, jejunum, ileum, caecum, colon and rectum were noted for all animals at macroscopic examination. This was considered due to the colour of the test material. At 300 mg/kg, no abnormalities were found.
Other findings:
The oral LD50 value was established to be within the range of 300 to 2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 500 mg/kg body weight.

Applicant's summary and conclusion

Interpretation of results:
other: Classified as Category 4 in accordance with EU criteria
Conclusions:
Under the conditions of the study, the oral LD50 value of the test material was established to be within the range of 300 to 2000 mg/kg body weight.
Executive summary:

The acute oral toxicity potential of the test material to the rat was investigated in accordance with the standardised guidelines OECD 423, EU Method B1 tris, US EPA OPPTS 870.1100 and JMAFF 12 Nousan Notification No 8147 under GLP conditions using the Acute Toxic Class Method.

Initially, the test material was administered by oral gavage to three female Wistar rats at 2000 mg/kg body weight. In a stepwise procedure two additional groups of three females were dosed at 300 mg/kg body weight. All animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).

At 2000 mg/kg, two animals were found dead on Day 1 and one animal was found dead on Day 2. At 300 mg/kg, no mortality occurred. At 2000 mg/kg, lethargy, hunched posture, rales, shallow respiration, salivation, ptosis and/or hypothermia were noted for all animals on Day 1. At 300 mg/kg, hunched posture and/or salivation were noted for all animals on Days 1. Yellow discoloration of the back, faeces, urine, forelegs, snout, paws, genital region and/or mouth was noted for most animals. This was considered due to the colour of the test material.

The mean body weight gain shown by the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain. At 2000 mg/kg, yellow discoloration of the contents of the stomach, duodenum, jejunum, ileum, caecum, colon and rectum were noted for all animals at macroscopic examination. This was considered due to the colour of the test material. At 300 mg/kg, no abnormalities were found.

Under the conditions of the study, the oral LD50 value of the test material in Wistar rats was established to be within the range of 300 to 2000 mg/kg body weight. In accordance with the OECD 423 test guideline, the LD50 cut-off value was considered to be 500 mg/kg body weight.