Registration Dossier

Administrative data

Description of key information

There are good quality GLP Guidelines studies for acute toxicity by the oral and dermal routes. The dermal route is the most likely route of exposure during the manufacture and use of the substance. These show a consistent pattern of low acute toxicity. Acute Inhalation testing has been waived due to the low toxicity by the other route and inhalation not been expected in the handling and use of this substance.

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:
From January 05, 2009 to January 30, 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Sex:
male/female
Details on test animals and environmental conditions:
Animals
Number of Animals: 9
Sex: Female, nulliparous and non-pregnant.
Species/Strain: Rat/Sprague-Dawley derived, albino.
Age/Body weight: Young adult (9-11 weeks)/160-225 grams at experimental start.
Source: Received from Ace Animals, Inc., Boyertown, PA on December 16 and 30, 2008 and January 13, 2009.

Husbandry
Housing: The animals were singly housed in suspended stainless steel caging with mesh floors which conform to the size recommendations in the most recent Guide for the Care and Use of Laboratory Animals DHEW (NIH). Litter paper was placed beneath the cage and was changed at least three times per week.
Animal Room Temperature and Relative Humidity Ranges: 20-21ºC and 15-60%, respectively. The low humidity reading was due to a temporary malfunction of the environmental control system.
Photoperiod: 12-hour light/dark cycle
Acclimation Period: 6-22 days
Food: Purina Rodent Chow #5012
Water: Filtered tap water was supplied ad libitum by an automatic water dispensing system.
Contaminants: There were no known contaminants reasonably expected to be found in the food or water at levels which would have interfered with the results of this study. Analyses of the food and water are conducted regularly and the records are kept in the study fecility.
Cage: Each cage was identified with a cage card indicating at least the study number, dose level, identification and sex of the animal.
Animal: A number was allocated to each rat on receipt and a stainless steel ear tag bearing this number was attached to the rat. This number, together with a sequential animal number assigned to study number 26611, constituted unique identification.
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
Selection of Animals
Prior to each dosing, experimentally naive rats were fasted overnight by removing the feed from their cages. During the fasting period, the rats were examined for health and weighed (initial). Nine healthy, naive female rats (not previously tested) were selected for test.

Dose Calculations
Individual doses were calculated based on the initial body weights, taking into account the specific gravity (determined by EPSL) of the test substance.

Dosing
The test substance was administered to the stomach using a stainless steel ball-tipped gavage needle attached to an appropriate syringe. Following administration, each animal was returned to its designated cage. Feed was replaced approximately 3-4 hours after dosing.
Initially, a single animal received a limit dose of 5000 mg/kg. Due to the mortality of this animal, a Main Test was conducted. For the Main Test, the test substance was administered to the animals in sequence described below in the table. The decision to proceed with the next animal was based on the survival of the previous animal following dosing.
Doses:
175 mg/kg; 550 mg/kg; 1,1750 mg/kg and 5,000 mg/kg
No. of animals per sex per dose:
Main study:
175 and 550 mg/kg Dose Levels (1 animal each); 1,750 mg/kg Dose Level (3 animals); 5,000 mg/kg Dose Level (4 animals)
Control animals:
no
Details on study design:
Body Weights
Individual body weights of the animals were recorded prior to test substance administration (initial) and again on Days 7 and 14 (termination) following dosing or after death.

Cage-Side Observations
The animals were observed for mortality, signs of gross toxicity, and behavioral changes during the first several hours post-dosing and at least once daily thereafter for 14 days after dosing or until death occurred (see Section 9). Observations included gross evaluation of skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity and behavior pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhea, and coma.

Necropsy
Surviving rats were euthanized via CO2 inhalation at the end of the 14-day observation period. Gross necropsies were performed on all decedents and euthanized animals. Tissues and organs of the thoracic and abdominal cavities were examined.
Statistics:
The Acute Oral Toxicity (Guideline 425) Statistical Program (Weststat, version 1.0, May 2001) was used for all data analyses including: dose progression selections, stopping criteria determinations and/or LD50 and confidence limit calculations.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
3 129 mg/kg bw
Based on:
test mat.
95% CL:
> 1 750 - < 5 000
Remarks on result:
other: female
Mortality:
175 and 550 mg/kg Dose Levels (1 animal each): both animals survived.
1,750 mg/kg Dose Level (3 animals): all animals survived test substance administration.
5,000 mg/kg Dose Level (4 animals): all animals died within three hours of test substance administration.
Clinical signs:
175 and 550 mg/kg Dose Levels (1 animal each): both animals appeared active and healthy during the study.
1,750 mg/kg Dose Level (3 animals): following administration, one animal was hypoactive and exhibited facial staining. All animals exhibited ano-genital staining and/or reduced fecal volume. However, these animals recovered by Day 3 and appeared active and healthy for the remainder of the 14-day observation period.
5,000 mg/kg Dose Level (4 animals): prior to death these animals were hypoactive and exhibited abnormal posture and piloerection.
Body weight:
1,750 mg/kg Dose Level (3 animals): all animals gained body weight during the study.
Gross pathology:
175 and 550 mg/kg Dose Levels (1 animal each): there were no signs of gross toxicity, adverse pharmacologic effects, or abnormal behavior. No gross abnormalities were noted for these animals when necropsied at the conclusion of the 14-day observation period.
1,750 mg/kg Dose Level (3 animals): no gross abnormalities were noted for the animals when necropsied at the conclusion of the 14-day observation period.
5,000 mg/kg Dose Level (4 animals): gross necropsy of the decedents revealed red intestines.
Interpretation of results:
GHS criteria not met
Conclusions:
Under the study conditions, the acute oral LD50 of the test substance was estimated to be 3129 mg/kg bw in female rats (based on an assumed sigma of 0.5) with an approximate 95% PL confidence interval of 1750 mg/kg (lower) to 5000 mg/kg (upper).
Executive summary:

A study was conducted to determine the potential for the test substance to produce toxicity from a single dose via the oral route according to OECD Guideline 425 and EPA OPPTS Method 870.1100, in compliance with GLP. An initial limit dose of 5000 mg/kg bw was administered to one healthy female rat by oral gavage. Due to mortality in this animal, the study proceeded to the main test using the default starting level of 175 mg/kg bw and following the up and down procedure, eight additional females were dosed at levels of 175, 550, 1750 or 5000 mg/kg bw. Females were selected for the test because they are frequently more sensitive to the toxicity of test compounds than males. All animals were observed for mortality, signs of gross toxicity, and behavioral changes at least once daily for 14 d after dosing or until death occurred. Body weights were recorded prior to administration and again on Days 7 and 14 (termination) following dosing or after death. Necropsies were performed on all animals. Under the study conditions, the acute oral LD50 of the test substance was estimated to be 3129 mg/kg bw in female rats (based on an assumed sigma of 0.5) with an approximate 95% PL confidence interval of 1750 mg/kg (lower) to 5000 mg/kg bw (upper) (Oley, 2010).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
3 129 mg/kg bw
Quality of whole database:
The test substance has acute oral toxicity estimated by the up and down method to be 3129 mg/kg bodyweight. The study is a modern GLP compliant OECD guideline study Klimsch 1.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

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:
From January 12, 2009 to January 26, 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
Number of Animals: 10
Sex: 5 Males and 5 Females. Females assigned to test were nulliparous and nonspregnant.
Species/Strain: Rats/Sprague-Dawley derived, albino.
Age/Body weight: Young adult (10-11 weeks)/males 359-382 grams and females 222-240 grams at experimental start.
Source: Received from Ace Animals, Inc., Boyertown, PA on December 18, 2008.
Housing: The animals were singly housed in suspended stainless steel caging with mesh floors, which conform to the size recommendations in the most recent Guide for the Care and Use of Laboratory Animals DHEW (NIH). Litter paper was placed beneath the cage and was changed at least three times per week.
Animal Room Temperature and Relative Humidity Ranges: 18-22ºC and 12-27%, respectively. The low humidity occurred during the study due to a temporary malfunction of the environmental control system.
Photoperiod: 12-hour light/dark cycle
Acclimation Period: 25 days
Food: Purina Rodent Chow #5012
Water: Filtered tap water was supplied ad-libitum by an automatic water dispensing system.
Contaminants: There were no known contaminants reasonably expected to be found in the food or water at levels, which would have interfered with the results of this study. Analyses of the food and water are conducted regularly and the records are kept in the laboratory facility.
Cage: Each cage was identified with a cage card indicating at least the study number and identification and sex of the animal.
Animal: A number was allocated to each rat on receipt and a stainless steel ear tag bearing this number was attached to the animal. This number, together with a sequential animal number assigned to study 26612, constituted unique identification.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
Preparation and Selection of Animals
On the day prior to application, a group of animals was prepared by clipping the dorsal area and the trunk. After clipping and prior to application, the animals were examined for health, weighed (initial), and the skin checked for any abnormalities. Ten healthy, naive rats (five males and five females; not previously tested) were selected for test.

Dose Calculations
Individual doses were calculated based on the initial body weights, taking into account the specific gravity (determined by EPSL) of the test substance.

Application of Test Substance
Five thousand mg/kg of body weight of the test substance was applied evenly over a dose area of approximately 2 inches x 3 inches (approximately 10% of the body surface) and covered with a 2-inch x 3-inch, 4-ply gauze pad. The gauze pad and entire trunk of each animal were then wrapped with 3-inch Durapore tape to avoid dislocation of the pad and to minimize loss of the test substance. The rats were then returned to their designated cages. The day of application was considered Day 0 of the study. After 24 hours of exposure to the test substance, the pads were removed and the test sites were gently cleansed of any residual test substance.
Duration of exposure:
24 hours.
Doses:
5000 mg/kg bw
No. of animals per sex per dose:
10
Control animals:
not required
Details on study design:
Body Weights
Individual body weights of the animals were recorded prior to test substance application (initial) and again on Days 7 and 14 (termination).

Cage-Side Observations
The animals were observed for mortality, signs of gross toxicity, and behavioral changes during the first several hours after application, and at least once daily thereafter for 14 days.
Observations included gross evaluation of skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity and behavior pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhea, and coma.

Necropsy
All rats were euthanized via CO2 inhalation on Day 14. Gross necropsies were performed on all animals. Tissues and organs of the thoracic and abdominal cavities were examined.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
5 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortalities were observed.
Clinical signs:
Following application, all animals exhibited facial staining and/or ano-genital staining between Days 1 and 2. Also, dermal irritation (erythema, edema, desquamation, hyperkeratosis, discoloration, eschar and/or corrosion) was noted at the dose site of all animals between Days 1 and 14.
Body weight:
All animals survived exposure to the test substance and gained body weight during the study.
Gross pathology:
Gross necropsy of the females revealed red intestines. No gross abnormalities were noted for any of the males when necropsied at the conclusion of the 14-day observation period.
Interpretation of results:
GHS criteria not met
Conclusions:
Under the study conditions, the acute dermal LD50 of the test substance was greater than 5000 mg/kg bw in male and female rats.
Executive summary:

A study was conducted to determine the potential acute toxicity of the test substance by the dermal route according to OECD Guideline 402, EU Method B.3 and EPA OPPTS Method 870.1200, in compliance with GLP. the test susbtance was applied a single time at 5000 mg/ kg bw to the skin of 10 healthy rats for 24 h. The animals were observed for mortality, signs of gross toxicity and behavioral changes at least once daily for 14 d. Body weights were recorded prior to application and again on Days 7 and 14 (termination). Necropsies were performed on all animals at terminal sacrifice. All animals survived exposure to the test substance and gained body weight during the study. Following application, all animals exhibited facial staining and/or ano-genital staining between Days 1 and 2. Also, dermal irritation (erythema, edema, desquamation, hyperkeratosis, discoloration, eschar and/or corrosion) was noted at the dose site of all animals between Days 1 and 14. Gross necropsy of the females revealed red intestines. No gross abnormalities were noted for any of the males when necropsied at the conclusion of the 14 d observation period. Under the study conditions, the acute dermal LD50 of the test substance was greater than 5000 mg/kg bw in male and female rats (Oley, 2010).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw
Quality of whole database:
Acute dermal toxicity, the dermal LD50 in rats is greater than 5000mg/kg bodyweight. There were no deaths at 5000mg/kg but there was dermal irritation/corrosion at this dose level. The study is a modern OECD guideline study fully GLP compliant. It is Klimisch 1.

Additional information

Justification for classification or non-classification

Both the oral and dermal toxicity showed low acute toxicity. In the acute oral toxicity study there were deaths at 5000 m/kg but no deaths or severe toxicity at 1750 mg/kg. Based on this the Oral LD was calculated to be 3129 mg/kg so this results in a classification by GHS criteria of category 5 for acute oral toxicity but it not classified for acute oral toxicity for (oral LD50 greater than 2000 mg/kg).

In the dermal acute toxicity study there were no deaths at the 5000 m/kg top dose so the dermal LD50 is greater than 5000 mg/kg. Therefore, there is no classification for acute dermal toxicity either according to GHS or EU CLP requirements.