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Description of key information

Acute oral toxicity: 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 50 to 300 mg/kg body weight.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 July 2016 to 16 August 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
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
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 - 10 weeks old)
- Weight at study initiation: 140 to 209 g (mean 167 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
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 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: Initially, the test material was administered by oral gavage to three rats at 2000 mg/kg body weight. In a stepwise procedure, three additional groups of three females were dosed at 300 and 50 mg/kg body weight (one and two groups, respectively).
Doses:
2000, 300 and 50 mg/kg body weight
No. of animals per sex per dose:
3 animals per sex per dose group (1, 1 and 2 groups for the 2000, 300 and 50 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. Animals showing pain, distress or discomfort, which was considered not transient in nature or was likely to become more severe, were sacrificed for humane reasons. Body weights were recorded on Days 1 (pre-administration), 8 and 15. 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. The moribund animals and animals surviving to the end of the observation period 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).
Preliminary study:
not perfomed
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 50 - < 300 mg/kg bw
Based on:
test mat.
Mortality:
At 2000 mg/kg, all animals were found dead or sacrificed for humane reasons on Day 1. At 300 mg/kg all, animals were found dead or sacrificed for humane reasons on Day 1. At 50 mg/kg, no mortality occurred.
Clinical signs:
At 2000 mg/kg, lethargy, flat posture, hunched posture, slow breathing, salivation, hypothermia and/or orange urine were noted for all animals on Day 1.
At 300 mg/kg, lethargy, flat posture, hunched posture, slow breathing, shallow respiration, piloerection, ptosis and/or purple (mouth) were noted for all animals on Day 1.
At 50 mg/kg, lethargy, hunched posture, rales, shallow respiration, piloerection, salivation, and/or ptosis were noted for all animals on Day 1. Purple discolouration of the snout, faeces and urine were noted for three out of six animals between Days 1 and 4; this discoloration is believed to be 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, general whole body purple discoloration and purple discolouration of the stomach contents were noted for all animals. This was believed to be due to the colour of the test material.
At 300 mg/kg, general whole body purple discoloration and purple discolouration of the contents of the stomach, duodenum, jejunum, ileum and colon were noted for all animals. This was believed to be due to the colour of the test material. Additionally, an accentuated lobular pattern of the liver was noted for one animal.
At 50 mg/kg, no abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
The oral LD50 value was established to be within the range of 50 to 300 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 200 mg/kg body weight.
Interpretation of results:
other: Classified as Category 3 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 50 to 300 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 three additional groups of three females were dosed at 300 (1 group) and 50 (2 groups) 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, all animals were found dead or sacrificed for humane reasons on Day 1. At 300 mg/kg all, animals were found dead or sacrificed for humane reasons on Day 1. At 50 mg/kg, no mortality occurred.

At 2000 mg/kg, lethargy, flat posture, hunched posture, slow breathing, salivation, hypothermia and/or orange urine were noted for all animals on Day 1. At 300 mg/kg, lethargy, flat posture, hunched posture, slow breathing, shallow respiration, piloerection, ptosis and/or purple (mouth) were noted for all animals on Day 1. At 50 mg/kg, lethargy, hunched posture, rales, shallow respiration, piloerection, salivation, and/or ptosis were noted for all animals on Day 1. Purple discolouration of the snout, faeces and urine were noted for three out of six animals between Days 1 and 4, this discoloration is believed to be due to the colour of the test item. The body weight gain shown by the animals over the study period was considered to be normal.

During necropsy, at 2000 mg/kg general whole body purple discoloration and purple discolouration of the stomach contents were noted for all animals. This was believed to be due to the colour of the test material. At 300 mg/kg, general whole body purple discoloration and purple discolouration of the contents of the stomach, duodenum, jejunum, ileum and colon were noted for all animals. This was believed to be due to the colour of the test material. Additionally, an accentuated lobular pattern of the liver was noted for one animal. At 50 mg/kg, no abnormalities were found at macroscopic post mortem examination.

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 50 to 300 mg/kg body weight. In accordance with the OECD 423 test guideline, the LD50 cut-off value was considered to be 200 mg/kg body weight.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
50 mg/kg bw
Quality of whole database:
One study is available, conducted in accordance with standardised guidelines under GLP conditions. The quality of the database is therefore considered to be high.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

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. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

Initially, the test material was administered by oral gavage to three female Wistar rats at 2000 mg/kg body weight. In a stepwise procedure three additional groups of three females were dosed at 300 (1 group) and 50 (2 groups) 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, all animals were found dead or sacrificed for humane reasons on Day 1. At 300 mg/kg all, animals were found dead or sacrificed for humane reasons on Day 1. At 50 mg/kg, no mortality occurred.

At 2000 mg/kg, lethargy, flat posture, hunched posture, slow breathing, salivation, hypothermia and/or orange urine were noted for all animals on Day 1. At 300 mg/kg, lethargy, flat posture, hunched posture, slow breathing, shallow respiration, piloerection, ptosis and/or purple (mouth) were noted for all animals on Day 1. At 50 mg/kg, lethargy, hunched posture, rales, shallow respiration, piloerection, salivation, and/or ptosis were noted for all animals on Day 1. Purple discolouration of the snout, faeces and urine were noted for three out of six animals between Days 1 and 4, this discoloration is believed to be due to the colour of the test item. The body weight gain shown by the animals over the study period was considered to be normal.

During necropsy, at 2000 mg/kg general whole body purple discoloration and purple discolouration of the stomach contents were noted for all animals. This was believed to be due to the colour of the test material. At 300 mg/kg, general whole body purple discoloration and purple discolouration of the contents of the stomach, duodenum, jejunum, ileum and colon were noted for all animals. This was believed to be due to the colour of the test material. Additionally, an accentuated lobular pattern of the liver was noted for one animal. At 50 mg/kg, no abnormalities were found at macroscopic post mortem examination.

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 50 to 300 mg/kg body weight. In accordance with the OECD 423 test guideline, the LD50 cut-off value was considered to be 200 mg/kg body weight.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance requires classification with respect to acute toxicity via the oral route as Category 3 (H301: Toxic if swallowed).