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Administrative data

Description of key information

In an acute toxicity study the substance was administered to nine female Wistar Han rats by oral gavage with three animals receiving 300 mg/kg bw followed by six animals receiving a dose of 2000 mg/kg bw (single administration). At 300 mg/kg bw, hunched posture and piloerection were noted for all animals between days 1 and 6. At 2000 mg/kg bw, hunched posture and piloerection were noted for all animals on Days 1 and/or 2. There were no mortalities or other signs of clinical toxicity, mean body weight gain was comparable with the control animals and no abnormalities found at macroscopic post-mortem examination.The oral LD50 is considered to be >2000 mg/kg bw.

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:
04 January 2019 - 19 February 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
yes
Remarks:
Deviations from the minimum level of daily mean target humidity occurred on three days during the study (lowest value was 36%). This deviation did not affect the integrity of the study, as it had no effect of clinical conditions of the animals.
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EC No 440, part B: "Acute Oral Toxicity, Acute Toxic Class Method"
Version / remarks:
2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: JMAFF Guidelines
Version / remarks:
2000, including the most recent revisions (January 2019)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Specific details on test material used for the study:
Batch (Lot) Number: AS455433
Expiry date: 01 November 2020 (retest date) (taken from label)
Physical Description: Colourless to light yellow viscous liquid
Purity/Composition: 98.5%
Storage Conditions: At room temperature protected from light

Additional information
Test Facility test item number: 209996/A


Species:
rat
Strain:
other: Wistar Han
Sex:
female
Details on test animals or test system and environmental conditions:
Test System.

Species: Rat
Strain: Crl: WI(Han)
Condition: Outbred, SPF-Quality
Source: Charles River Deutschland, Sulzfeld, Germany
Age at the Initiation of Dosing: Young adult animals (approximately 8-10 weeks old) were selected.
Weight at the Initiation of Dosing: 151 to 179 g.


Housing.

On arrival and following assignment to the study, animals were group housed (up to 3 animals of the same sex and same dosing group together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles. The room in which the animals were kept was documented in the study records.
Animals were separated during designated procedures/activities. Each cage was clearly labeled.

Environmental Conditions.

Target temperatures of 18 to 24°C with a relative target humidity of 40 to 70% were maintained. The actual daily mean temperature during the study period was 20 to 21°C with an actual daily mean relative humidity of 36 to 52%. A 12 hour light/12 hour dark cycle was maintained. Ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained in the animal rooms.


Selection, Assignment, Replacement, and Disposition of Animals.

Animals were assigned to the study at the discretion of the coordinating biotechnician according to body weights, with all animals within ± 20% of the sex mean. Animals in poor health or at extremes of body weight range were not assigned to the study.
Before the initiation of dosing, a health inspection was performed and any assigned animal considered unsuitable for use in the study were replaced by alternate animals obtained from the same shipment and maintained under the same environmental conditions.
The disposition of all animals was documented in the study records.

Food.

Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures.
The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility.
It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.

Water.

Municipal tap-water was freely available to each animal via water bottles.
Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility.
It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.

Animal Enrichment.

For psychological/environmental enrichment, animals were provided with paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom), except when interrupted by study procedures/activities.

Veterinary Care.

Veterinary care was available throughout the course of the study; however, no examinations or treatments were required.
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
A single dose of substance was administered to the appropriate animals by oral gavage on Day 1, using a syringe with a plastic gavage cannula attached.
The dose volume for each animal was based on the body weight measurement prior to dosing. Dose volume (mL/kg body weight) was calculated as follows:
Dose level (g/kg) / spec.gravity or density (g/mL).
The substance was stirred continuously during dose administration.
Animals were deprived of food overnight (for a maximum of 20 hours) prior to dosing and until 3-4 hours after administration of the substance. Water was available.
Doses:
Single dose of 300 and 2000 mg/kg body weight.
No. of animals per sex per dose:
Three animals for 300 mg/kg bw
In a stepwise procedure two additional groups of three females were dosed at 2000 mg/kg body weight.
Control animals:
no
Details on study design:
Justification of Route and Dose Levels.

The oral route was selected as it is a possible route of human exposure during manufacture, handling or use of the substance.
The dose levels were based on the OECD test guidelines and were selected from the series 5 (lowest dose level), 50, 300 and 2000 (highest dose level) mg/kg body weight. The starting dose level should be the one that is likely to produce mortality in at least some of the animals and was selected based on available toxicity data of the substance.

Experimental Design.

The toxicity of the substance was assessed by stepwise treatment of groups of 3 females. 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. The first group was treated at a dose level of 300 mg/kg. Based on the results, two additional groups were dosed at 2000 mg/kg.
Throughout the study, animals were observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day. Animals were not removed from cage during observation, unless necessary for identification or confirmation of possible findings.

Clinical Observations:

1) Postdose Observations.

Postdose observations were performed at periodic intervals on the day of dosing (at least three times) and once daily thereafter. The observation period was 14 days.
All the animals were examined for reaction to dosing. The onset, intensity and duration of these signs was recorded (if appropriate). Signs were graded for severity and the maximum grade was predefined at 3 or 4. Grades were coded as slight (grade 1), moderate (grade 2), severe (grade 3) and very severe (grade 4). For certain signs, only its presence (grade 1) or absence (grade 0) was scored.

2) Body Weights.

Animals were weighed individually on Day 1 (predose), 8 and 15. A fasted weight was recorded on the day of dosing.

3) Terminal Procedures.

All animals were sacrificed by oxygen/carbon dioxide procedure at the end of the observation period. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded.
Statistics:
Not applicable.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality occured
Remarks:
At 300 mg/kg, hunched posture and piloerection were noted for all animals between Days 1 and 6. At 2000 mg/kg, hunched posture and piloerection were noted for all animals on Days 1 and/or 2.
Mortality:
None
Clinical signs:
other: At 300 mg/kg bw, hunched posture and piloerection were noted for all animals between Days 1 and 6. At 2000 mg/kg bw, hunched posture and piloerection were noted for all animals on Days 1 and/or 2.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.

Clinical Observation

Sex

Dose

ID

Administration Result

Clinical Observation

300 mg/kg b. w.

1

alive

no signs of intoxication, change of health, nor any other adverse reactions during 24 hours and 15-days observation period

2

alive

no signs of intoxication, change of health, nor any other adverse reactions during 24 hours and 15-days observation period

3

alive

no signs of intoxication, change of health, nor any other adverse reactions during 24 hours and 15-days observation period

Hunched posture and piloerection were noted for all animals between Days 1 and 6. 

Sex

Dose

ID

Administration Result

Clinical Observation

2000 mg/kg b. w.

4

alive

no signs of intoxication, change of health, nor any other adverse reactions during 24 hours and 15-days observation period

5

alive

no signs of intoxication, change of health, nor any other adverse reactions during 24 hours and 15-days observation period

6

alive

no signs of intoxication, change of health, nor any other adverse reactions during 24 hours and 15-days observation period

7

alive

no signs of intoxication, change of health, nor any other adverse reactions during 24 hours and 15-days observation period

8

alive

no signs of intoxication, change of health, nor any other adverse reactions during 24 hours and 15-days observation period

9

alive

no signs of intoxication, change of health, nor any other adverse reactions during 24 hours and 15-days observation period

Hunched posture and piloerection were noted for all animals on Days 1 and/or 2.

Body Weight

Sex

Dose

ID

Body Weight (g)

Day 1

Day 8

Day 15

300 mg/kg b. w.

1

156

182

183

2

168

193

196

3

175

200

208

Sex

Dose

ID

Body Weight (g)

Day 1

Day 8

Day 15

2000 mg/kg b. w.

4

155

173

181

5

179

200

216

6

163

193

201

7

153

189

199

8

151

182

193

9

151

186

201

Interpretation of results:
GHS criteria not met
Remarks:
LD50 > 2000 mg/kg body weight
Conclusions:
The oral LD50 in an acute toxicity study conducted on the substance in female rats administered via gavage is >2000 mg/kg bw.
Executive summary:

In a reliable in vivo acute oral toxicity study, the substance was administered to 9 female Wistar Han rats in a single dose by oral gavage. An initial dose of 300 mg/kg body weight was administered to 3 rats. Based on these results, two additional groups of three females were dosed at 2000 mg/kg body weight in a stepwise procedure. All (6/6 females) animals survived the dose of 2000 mg/kg bw. At 300 mg/kg bw, hunched posture and piloerection were noted for all animals between days 1 and 6. At 2000 mg/kg bw, hunched posture and piloerection were noted for all animals on Days 1 and/or 2. During the observation period of 15 days, no other signs of intoxication, change of health, nor any other adverse reactions were seen. Macroscopic examination of the animals at the end of the study did not reveal treatment-related changes in any of the treatment groups. The LD50 of the substance is considered to be greater than 2000 mg/kg body weight after a single oral administration to female Wistar Han rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Sufficient to address requirements

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

Justification for classification or non-classification

Based on the findings of a reliable acute oral toxicity study conducted on the substance, classification of the substance is not justified.

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