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Description of key information

Three acute oral toxicity studies are available for the Substance. The acute oral toxicity study conclusions are summarised as follows:

Acute Oral 2016 - LD50 > 2000 mg/kg

Acute Oral 2000 - LD50 > 2000 mg/kg

Acute Oral 1976 - LD50 > 5000 mg/kg

Based on the weight of evidence the data indicates that the estimated LD50 for the substance is > 5000 mg/kg

One acute dermal toxicity studies are available for the Substance. The acute dermal toxicity study concluded that the Acute dermal LD50 was > 3160 mg/kg

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30 October 2015 - 13 April 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)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Specific details on test material used for the study:
Identification: EXP1505385
Label Lot No.: E00350-288
Batch No: E00974-38D&F
Re-test Date: 22 June 2018*
Physical Description: Pale amber waxy solid
Purity: 100 %
Storage Conditions: Ambient temperature/ Dark
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl:CD (SD) (
Sex:
female
Details on test animals and environmental conditions:
Twelve female Sprague-Dawley Crl:CD (SD) (nulliparous and non-pregnant) rats were supplied by Charles River, Margate, UK and arrived at the Test Facility on 27 Oct2015. Nine rats were used on this study and the remaining 3 animals (scheduled for treatment in Group 4) were not used and returned to stock after completion of the study. Animals were approximately 7 to 8 week old on despatch and weighed between 176 and 200 g. On the day of dosing animals were approximately 8 to 11 weeks old and weighed between 180 and 229 g. All animals weighed within +/- 20% of the mean body weight of the previously dosed groups.

The animals were allowed to acclimatise to the Test Facility rodent toxicology accommodation for a period of at least 8 days before dosing.

Animals were housed in groups of 3 in appropriately sized solid-bottomed polycarbonate cages with stainless steel mesh tops. Wood shavings were used as bedding.
Husbandry practices and environmental enrichment were in accordance with Test Facility SOPs and the protocol. SDS Rat and Mouse (modified) No. 1 Diet SQC Expanded was available ad libitum. Food was withdrawn overnight before dosing, and returned as soon as practicable after administration of the test item. Water from the public supply was available ad libitum.

Certificates of analysis for significant contaminants in diet, water, bedding and environmental enrichment materials are retained at the Test Facility. There were no contaminants present in significant quantities that would have interfered with the objectives of the study.
The environmental conditions were continually monitored and recorded every 15 min. Target ranges were 19¿C to 23°C for temperature and 40% to 70% for relative humidity. From animal arrival, the temperature was between approximately 19¿C and 23¿C and the relative humidity was approximately 31% to 74%. On occasions where the target environmental conditions were not mentioned the change was transient and did not trigger the internal alarm and notification procedure at the Test Facility. The health of the animals was unaffected in any occasion, and these events were considered not to have had an impact on the outcome or integrity of the study.

A 12 h light/dark cycle was in operation (light hours 0700-1900 h) with a minimum of 10 air changes per hour.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
The animals received a single oral dose via a gavage tube after being deprived of food overnight. The doses were calculated on the basis of each animal’s body weight on the day of dosing.
Doses:
Dose level of 300 mg/kg and 2000 mg/kg were used
No. of animals per sex per dose:
3 animals were tested at 300 mg/kg
6 animals were tested at 2000 mg/kg
Control animals:
no
Details on study design:
A dose of 300 mg/kg was selected as a suitable starting dose level for administration to Group 1 animals. There were no adverse signs in these animals so 2000 mg/kg was chosen as dosage level for Group 2, and as there were no unscheduled deaths, the third group of rats was treated at the same dose level to complete the study.

All animals were for checked for mortality early each morning and as late as possible each day for viability. All animals were examined for clinical signs to treatment for up to 5 hours post dosing on the day of dosing and once daily thereafter, until Day 15. The body weight of each individual animal was recorded before dosing on Day 1 and on Days 8 and 15.

Animals were euthanised on Day 15 by exposure to a rising concentration of carbon dioxide followed by exsanguination. Gross necropsy was performed on all animals. The necropsy consisted of an examination of the cranial, thoracic and abdominal organs and tissues in situ.
Statistics:
No formal statistical analysis was conducted.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no unscheduled deaths.
Clinical signs:
There were no adverse signs noted at 300 mg/kg. After treatment at 2000 mg/kg adverse clinical signs were noted from 15 minutes after dosing up to 3 days after dosing (Day 4). The majority of adverse signs were recorded 30 minutes to 2 hours after dosing, and included decreased activity, eyes partly closed, piloerection (fur erect), a hunched appearance and abnormal gait. There were wet muciod feaces between 3-4 hours after dosing. The animals gradually recovered and 3 days after dosing only one animal still had a hunched appearance. There were no adverse signs from Day 5.
Body weight:
Slight body weight loss of approximately 2% in 3 of the animals that received 2000 mg/kg, when body weights recorded on Day 8 were compared with those on Day 15. The body weight gains of animals at 300 or 2000 mg/kg were generally considered to be acceptable for rats of this age and strain.
Gross pathology:
There were no macroscopic abnormalities in any animal.
Other findings:
Not Applicable

See attached background document

Interpretation of results:
GHS criteria not met
Conclusions:
The median lethal dose level (LD50) of Substance was considered to be > 2000 mg/kg.
Executive summary:

The objective of this study was to assess the adverse effects which can follow within a short period of time after a single oral (gavage) administration of the test item, Substnace to rats.

The test item was administered to 3 groups of female Sprague- Dawley rats. The study design was as follows:

Text Table1
Experimental Design

Group
Numbers

Number of Animals

Dose Level
(mg/kg)

Dose Volume
(mL/kg)

Dose
Concentration (mg/mL)

1

3

300

10

30

2

3

2000

10

200

3

3

2000

10

200

The following parameters and end points were evaluated in this study: clinical signs, body weights, body weight changes, and on Day 15, gross necropsy findings.

There were no unscheduled deaths. There were no adverse signs noted at 300 mg/kg. After treatment at 2000 mg/kg adverse clinical signs were noted from 15 minutes after dosing up to 3 days after dosing (Day 4). The majority of adverse signs were recorded 30 minutes to 2 hours after dosing, and included decreased activity, eyes partly closed, piloerection (fur erect), a hunched appearance and abnormal gait. There were wet muciod feaces between 3-4 hours after dosing. The animals gradually recovered and 3 days after dosing only one animal still had a hunched appearance. There were no adverse signs from Day 5. Slight body weight loss of approximately 2% in 3 of the animals that received 2000 mg/kg, when body weights recorded on Day 8 were compared with those on Day 15. The body weight gains of animals at 300 or 2000 mg/kg were generally considered to be acceptable for rats of this age and strain. There were no macroscopic abnormalities in any animal.

Under the conditions of the study the median lethal dose level (LD50) of test Substance in Sprague-Dawley rats was considered to exceed 2000 mg/kg.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
10 September 1976
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
abstract
Justification for type of information:
Limited scopes for methodological differences with an acute oral study, therefore should be suitable to be used as supporting data and add to the weight of evidence of acute oral toxicity.
Qualifier:
no guideline available
Principles of method if other than guideline:
Ten males rats were treated orally with the test Substance at 5000 mg/kg and observed for 7 Days.
GLP compliance:
not specified
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
Physical Description: Brown Solid
Species:
rat
Sex:
male
Details on test animals and environmental conditions:
No data
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
Heated gently in a water bath and dosed as a warm liquid
Doses:
5000 mg/kg
No. of animals per sex per dose:
10 animals treated at 5000 mg/kg
Control animals:
not specified
Details on study design:
Test material was heated gently in a water bath and was dosed as a warm liquid to 10 animals at 5000 mg/kg. Animals were observed for 7 days for signs or mortality and clinical signs.
Statistics:
Not applicable, limit test
Sex:
male
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Mortality:
No mortality was observed in any animal during the observation period.
Clinical signs:
Hypo-activity was exhibited by all the animals within 5 minutes post-oral administration of the test substance. Diarrhoea was exhibited by all the animals at 1 hour. These symptoms had subsided by the following morning.
Body weight:
There were no body weight effects observed from test day 0 to Test Day 7.
Gross pathology:
No data
Interpretation of results:
other: Supportive of Key Study result
Conclusions:
No mortality was observed in any animal following treatment of the test Substance at 5000 mg/kg. Therefore the LD50 is considered to be > 5000 mg/kg.
Executive summary:

Ten males rats were treated orally with the test Substance at 5000 mg/kg and observed for 7 Days.

No mortality was observed in any animal during the observation period.

Hypo-activity was exhibited by all the animals within 5 minutes post-oral administration of the test substance. Diarrhoea was exhibited by all the animals at 1 hour. These symptoms had subsided by the following morning.

The median lethal dose level (LD50) of Substance was considered to be > 5000 mg/kg.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 February 2000 - 21 May 2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
Date Received: September 13, 1999
Expiration Date: September 2004
Description: Amber solid
Storage Condition: Room temperature
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
More animals than required for the conduct of the study were purchased and acclimated. Animals determined to be unsuitable for inclusion in the study because of poor health, outlying body weight, or other abnormalities were excluded from selection by the Study Director and/or technical staff. Study animals were selected from the remaining animals using a computer-generated, body weight sorting program. Weight variation for individual animals was within ±20% of the mean body weight of their sex.

Room: PE1O1
Housing: Single housed during the study period.
Caging: Suspended stainless steel and wire mesh with absorbent paper below cages.
Feed: PMI Certified Rodent Diet Meal 5002, ad libitum
Manufacturer: PMI Feeds, Inc., Richmond, Indiana.
Analysis: Performed by PMI Feeds, Inc. Copies of the feed analyses are maintained at the testing laboratory.
Contaminants: There were no known contaminants in the feed believed to have been present at levels that may have interfered with this study.
Availability of feed: Checked at least once daily for all animals.
Water: Automatic Watering System, ad libitum
Supplier: Site well and potable water system.
Analysis: Periodic analysis is the responsibility of the testing laboratory. A copy of the results is maintained at the testing
laboratory.
Contaminants: There were no known contaminants in the water believed to have been present at levels that may have interfered with this study.
Availability of water: Checked at least once daily for all animals.
Temperature: 64 to 72 degrees Fahrenheit
Humidity: 30 to 70 percent relative humidity
Lighting: Approximately 12 hours light (0700 to 1900 hours) and 12 hours dark (1900 to 0700 hours) by automatic timer.
Environmental conditions: Monitored twice per day Monday through Friday and once daily on Saturday, Sunday and holidays. Additionally, a non GLP-compliant system monitored conditions continuously.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
The dose volume (8.0 mlfkg) was calculated by dividing the dose level (2 gfkg) by the concentration (0.25 g/ml). The dose volume was then multiplied by the body weight to arrive at the calculated dose.

The test substance was administered (Day 0, 0930-0941 hours) as a single oral intubation via syringe and a stainless steel, straight, ball tipped feeding needle.
Doses:
Limit dose of 2000 mg/kg
No. of animals per sex per dose:
Five males and 5 females
Control animals:
no
Details on study design:
This study was conducted in order to assess the acute oral toxicity and to estimate the LD50 of the test substance in the rat when administered orally as a single dose with a 14 day observation period.

After recording animal body weights, all food was removed from the animal cages at approximately 4:00 p.m. during the evening immediately prior to the day of administration of the test substance. Food was withheld until completion of dosing the following morning.

The test substance was heated to 75°C to melt it. The appropriate amount of corn oil was mixed into the test substance and the mixture was heated to75°C to facilitate the mixing. The mixture was then allowed to cool to 50°C for dosing.

The animals were examined for viability twice daily Monday through Friday, and once daily on weekends. Clinical observations were made as to the nature, onset, severity, and duration of toxicological signs at 1, 2, 4, and 6 hours after dosing, and once per day thereafter for a total of 14 days. Body weights were recorded on the day prior to dosing (pretest), the day of dosing (Day 0), on Day 7, and on Day 14.

After the Day 14 observations, all animals were weighed and sacrificed by exsanguination following CO2 asphyxiation.
A gross necropsy that included an examination of the external surface of the body, all orifices, the cranial, thoracic, and abdominal cavities and their contents was performed on all animals.
Statistics:
Statistical analyses included means and standard deviations of body weight and body weight change by group and sex (Snedecor and Cochran, 1989).
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
No mortality was observed
Clinical signs:
The only clinical signs observed during the study were soft stool, mucoidal stool, and staining of the ano genital area during the 1, 2, 4, and 6 Hour-observations and at the Day 1 observations. Staining of the ano-genital area and mucoidal stool appeared first in four female rats at the 1-Hour observation. The females and males were observed with the same signs and/or soft stool at later observations during Day 0. On Day 1
two females were noted with staining of the ano-genital area. No clinical signs were observed after the Day 1 observation.
Body weight:
All animals displayed increases in body weight over their initial values.
Gross pathology:
The only gross postmortem observation was mottled lungs in two male rats and two female rats. All other animals were free of gross postmortem abnormalities.

See attached background documents

Interpretation of results:
GHS criteria not met
Remarks:
Based on weight of evidence
Conclusions:
The median lethal dose level (LD50) of Substance was considered to be > 2000 mg/kg.
Executive summary:

The acute oral toxicity of the test substance was evaluated when administered as a single dose via oral intubation at a limit dose of 2000 mg/kg to 5 male and 5 female Crl:CD®(SD)IGSBR rats. Clinical observations were made 1, 2, 4 and 6 hours after dose administration and once daily for 14 days. Body weights were recorded for all animals the day prior to dosing (pretest), the day of dosing (Day 0), Day 7, and Day 14. On Day 14, all animals were sacrificed and gross necropsies were perfonned.

All animals survived to scheduled termination on Day 14 and displayed increases in body weight over their initial values. Two male rats and one female rat were free of clinical signs during the study. The only clinical signs observed during the study were soft stool, mucoidal stool, and staining of the ano-genital area during Day Oil. No clinical signs were noted after the Day 1 observations. The only gross postmortem observation was mottled lungs in four rats.

In conclusion, oral intubation of the test substance at a dose level of 2000 mg/kg did not produce mortality or overt signs of toxicity under conditions of this study. Based on these results, the acute oral LD50 of the test substance was greater than 2000 mg/kg (the limit dose) in male and female rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw
Quality of whole database:
Two studies were considered as reliable without restriction as it was conducted on the registered substance (as defined in section 1.1) and meets acceptable scientific standards. One study was considered to be supportive and suitable to be used as part of a weight of evidence.

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10 September 1976
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation
Justification for type of information:
Limited scopes for methodological differences with an acute dermal toxicitystudy, therefore should be suitable to be used. As the study has been conducted on the susbtance and the toxicity profile is consistent, to prevent use of unnecessary animal testing this data should be considered and used.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
no
Specific details on test material used for the study:
Physical Description Brown Solid
Test material was grounded and applied as an aqeous slurry
Species:
rabbit
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
No Data
Type of coverage:
not specified
Vehicle:
not specified
Details on dermal exposure:
The test skin of all animals were abraded
Doses:
200, 794, 3160 mg/kg
No. of animals per sex per dose:
2 males and 2 females per dose
Control animals:
no
Details on study design:
Animals were treated dermally as an aqeous paste made from grounded test substance. Animals were observed for 14 Days for signs of mortality and clinical signs. Macrospic evaluation was performed at the end of the study.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 3 160 mg/kg bw
Based on:
test mat.
Mortality:
Motality was observed in 1/4 animals at 794 and at 3160 mg/kg
Clinical signs:
The material was severely irritating to the skin was observed through the observation period. Pale red to red, well-defined erythema, moderate edema and second degree burns at 200 mg/kg were observed. Pale red to red, well-defined erythema, severe edema and second degree burns at 794 and 3160 mg/kg were observed. Escharosis and fissuring were noted at all test sites of all survivors at 7 and 14 days.
Body weight:
Decreases in Bodyweight effects were noted in surviving 3 animals treated at 3160 mg/kg at observation Day 7. By day 14 animal bodyweights had partially recovered back to starting weights in 2 animals. One animal's bodyweight remained similar to Day 7.
Gross pathology:
No test related findings.

Dose Level (mg/kg)

Animal Number and Sex

Mortality

200

1-M

0/4

2-M

3-F

4-F

794

5-M

1/4

6-M

7-F

8-F

3160

9-M

1/4

10-M

11-F

12-F

Interpretation of results:
study cannot be used for classification
Conclusions:
LD50 = 3160 mg/kg
Executive summary:

The objective of this study was to assess the adverse effects which can follow within a short period of time after a single dermal application of the Substanc.

 

The Substance was administered to 3 groups of male and female, albino rabbits. The study design was as follows:

 

Table1
Experimental Design

 

Group
Numbers

Number of Animals

Dose Level
(mg/kg)

1

2 Males & 2 Females

200

2

2 Males & 2 Females

794

3

2 Males & 2 Females

3160

 

The following parameters and end points were evaluated in this study: clinical signs, body weights, body weight changes, and on Day 14, gross necropsy findings.

 

Mortality was observed in 1 animal treated at 3160 mg/kg bw day and 1 animal treated at 794 mg/kg bw day. Decreases in Bodyweight effects were noted in surviving 3 animals treated at 3160 mg/kg at observation Day 7. By day 14, animal bodyweights had partially recovered back to starting weights in 2 animals. One animal's bodyweight remained similar to Day 7.

The material was severely irritating to the skin was observed through the observation period. Pale red to red, well-defined erythema, moderate oedema and second degree burns at 200 mg/kg were observed. Pale red to red, well-defined erythema, severe oedema and second degree burns at 794 and 3160 mg/kg were observed. Escharosis and fissuring were noted at all test sites of all survivors at 7 and 14 days. No macroscopic findings were considered to be test related findings.

 

Under the conditions of the study the median lethal dose level (LD50) of the Substance in albino rabbits were considered to be 3160 mg/kg.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
3 160 mg/kg bw
Quality of whole database:
The study was considered as reliable with restriction as it was conducted on the registered substance (as defined in section 1.1) and meets acceptable scientific standards, but before the implementation of GLP and of the OECD Testing Guideline.

Additional information

Justification for classification or non-classification

The acute oral median lethal dose (LD50) of the Substance was estimated to be greater than 5,000 mg/kg body weight. The test substance did not meet the criteria for classification according to Regulation (EC) No.1272/2008 on the Classification, Labelling and Packaging of Substances and Mixtures.

The acute dermal median lethal dose (LD50) of the Substance was estimated to be 3,160 mg/kg body weight. The test substance did not meet the criteria for classification according to Regulation (EC) No.1272/2008 on the Classification, Labelling and Packaging of Substances and Mixtures.