Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Acute toxicity oral 
The acute oral LD50 for this test substance was calculated to be greater than 2000 mg/kg for male rats and 1414 mg/kg for female rats.
Acute toxicity inhalation
The four-hour LC50 values for this test substance were geater than 4.48 mg/L (highest attainable concentration) for both male and female rats.  
Acute toxicity dermal 
The acute dermal LD50 for this test substance was greater than 2000 mg/kg for male and female rats.

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:
1997-10-30 to 1997-11-24
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and guideline compliant study
Qualifier:
according to guideline
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Stone Ridge (Kingston), NY
- Age at study initiation: 7- 10 weeks
- Weight at study initiation: males: 187 - 203 g; females: 150 -189 g
- Housing: singly, in stainless-steel, wire mesh cages
- Diet: Certified Rodent Diet (PMI #5002, pelleted) was available ad libitum.
- Water: municipal drinking water, regularly assayed
- Acclimation period: 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 22
- Humidity (%): 48 - 55 %
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
Initially, a Iimit dose of 2000 mg/kg bw of the test substance was selected as the dose Ievel for the oral toxicity study. Based on results of this dose Ievel, additional dose Ievels of 1000 and 500 mg/kg bw of the test substance were selected for female rats.
Doses:
2000 mg/kg bw for male and female rats, 1000 and 500 mg/kg for female rats only
No. of animals per sex per dose:
2000 mg/kg bw dose group: 5 males and females
1000 mg/kg bw dose group: 5 females
500 mg/kg bw dose group: 5 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: three times on the day of administration, at least once each workday
- Observations included, but were not limited to examination of the hair, skin, eyes, mucous membranes, motor activity, feces, urine, respiratory systern, circulatory system, autonomic nervous system, central nervous system, and behavior patterns.
- Necropsy of survivors performed: yes, any animal that died during the study was necropsied as soon as possible. All surviving animals were euthanatized and necropsied at the completion of the 14-day observation period.
Statistics:
not required
Sex:
male
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Sex:
female
Dose descriptor:
LD50
Effect level:
1 414 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortalities were observed for the male animals.
Four of five female animals died following the administration of 2000 mg/kg bw.
No animal died follwing the administration of 1000 and 500 mg/kg bw.
Clinical signs:
other: Abnormal clinical signs were limited to reduced feces on the day following dosing from the male and female rats in the 2000 mg/kg bwdose group which survived to Day 1
Gross pathology:
Treatment-related changes observed at necropsy for the 2000 mg/kg bw female rats that died within 24 hours of test substance administration consisted of distention of the stomach with mucus, and necrosis and hemorrhage of the glandular gastric mucosa. No treatment-related changes were observed at necropsy for animals that survived the 14-day observation period. Incidental findings consisted of uterine hydrometra for single female rats from the 1000 and 500 mg/kg bw dose groups.
Other findings:
none
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 414 mg/kg bw
Quality of whole database:
The conservative LD50 for females was reported as key value. The LD50 for males was determined to be > 2000 mg/kg. The combined LD50 (male and female) was determined to be greater than 2000 mg/kg bw. GLP and guideline compliant study.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2000-01-06 until 2000-0218
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and guideline compliant study.
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
1981
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Ltd, Stone Ridge (Kingston), NY
- Age at study initiation: 46 - 49 days
- Weight at study initiation: males: 229 (+/-7) g; females: 163 (+/-5) g
- Housing: singly, suspended, stainless-steel, wire mesh cages
- Diet: Certified Rodent Diet (Purina Roden Chow #5002, pellets, ad libitum
- Water: drinking water, ad libitum, regulary analysed
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20.9- 24.3°C
- Humidity: 41.4-55.2 %
- Photoperiod: 12 hours of artificial light

Route of administration:
inhalation
Type of inhalation exposure:
nose/head only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: past nose-only units (CH Technologies, Westwood, NJ)
- Method of holding animals in test chamber: Restraining tubes
- Source and rate of air: hight dose groups: filtered, conditioned, exterior air; mid and low dose groups: filtered compressed air, flow rate of 8.6 - 10.6 L/min
- Method of conditioning air: filtering
- System of generating vapour: evaporation - airflow above surface of the test substance
- Temperature, humidity, pressure in air chamber: 19.0 - 26.0°C, 43 - 75%, ambient pressure, respectively
- Air flow rate: 8.6 - 10.6 L/min
- Air change rate: 86 to 106 air changes per hour
- Method of particle size determination: Micro Laser Particle Counter (model μLPC-301, Particle Measuring Systems, Inc., Boulder, CO)

TEST ATMOSPHERE
- Brief description of analytical method used: GC/FID and multipositional air sampling and analysis system.
- Samples taken from breathing zone: yes

Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
4.48 mg/L (highest attainable concentration - analytically verified)
2.5 mg/L (analytically verfified)
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
Observations
Rats were observed hourly during exposure for clinical signs of toxicity. However, due to the restraint of the animals in tubes, the during-exposure observations were limited to changes in respiration, eyes, and mucous membranes. Clinical examinations (were conducted before and after exposure and on each subsequent morning. Moribundity and mortality observations were conducted on subsequent weekday afternoons. On weekends, afternoon observations were not performed. Observations included, but were not limited to, examination of the hair, skin, eyes, mucous membranes, motor activity, feces, urine, respiratory system, circulatory system, autonomic nervous system, central nervous system, and behaviour patterns.

Body Weight Determinations
Body weights were measured on Days 0 (pre-exposure), 7, and 14.

Blood Collection and Euthanasia
Animals were fasted overnight beginning after the last exposure. The following day, animals were anesthetized with Isoflurane and blood was collected from the posterior vena cava. The blood was placed into vacutainer tubes and allowed to clot for analyses of serum. Other tubes containing an anticoagulant were used for analyses of whole blood samples. Blood smears were also prepared for blood cell counts. Following blood collection, the animals were euthanatized by exsanguination. Animals were bled and euthanatized in a random order based on a computer-generated list.

Necropsy
At the completion of the 14-day observation period, all animals were anesthetized with isoflurane, exsanguinated by severing the posterior vena cava, and necropsied.
Statistics:
Not applicable
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 4.48 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
No mortalities occurred.
Clinical signs:
other: On the day of exposure (Day 0) for the high-concentration group, one male rat had minor porphyrin discharges around the nose, minimal to moderate red discoloration of the hair was observed for two male and two female rats, and minimal to minor unkempt hai
Body weight:
All animals gained weight during both weeks of the study.
Gross pathology:
No test substance-related changes were observed at necropsy, and no tissues were collected for microscopic examination.
Other findings:
none
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating conc.
Value:
4 480 mg/m³ air
Quality of whole database:
GLP and guideline compliant study

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:
1997-11-11 until 1997-11-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Scientifically valid study according to GLP and current OECD guideline.
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
, adopted 1987
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
, adopted 1992
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Ltd, Hollister, CA
- Age at study initiation: 8 - 11 weeks
- Weight at study initiation: males: 273 - 297 g; femals: 224-252 g
- Housing: singly, suspended, stainless-steel, wire mesh cages
- Diet: Certified Rodent Diet (Purina Roden Chow #5002, pellets, ad libitum
- Water: drinking water, ad libitum, regulary analysed
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20-22°C
- Humidity: 49-50 %
- Photoperiod: 12 hours of artificial light

Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsal skin region
- % coverage: not less than 10 %
- Type of wrap if used: fiber pad and an occlusive wrap.

REMOVAL OF TEST SUBSTANCE
- Washing: using running water
- Time after start of exposure: 24 hours

TEST MATERIAL AND VEHICLE
The test substance was administered at a dose level of 2000 mg/kg bw

Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
10 animals: 5 males, 5 females
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: systemically between days 0-14, body weights were recorded on day 0, 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, organ weights, histopathology
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No adverse effects noted
Mortality:
No mortality was observed.
Clinical signs:
other: There were no signs of systemic reaction to treatment. There were no signs of systemic toxicity.
Gross pathology:
No abnormalities were noted.
Other findings:
none
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
GLP and guideline compliant study

Additional information

Acute toxicity oral

An acute oral toxicity study was conducted according to OECD guideline 401 in which male rats were administered 2000 mg/kg bw and female rats were administered 2000, 1000, and 500 mg/kg bw of the test substance by gavage. No mortality was observed for the animals of the male dose group. Four female rats assigned to the 2000 mg/kg dose group died within 24 hours of dosing. Clinical signs observed during the 14-day observation period were limited to reduced feces from the surviving 2000 mg/kg bw animals on the day following dosing. All surviving 2000 mg/kg bw animals appeared clinically normal by Day 2. Animals of the 1000 and 500 mg/kg dose group appeared clinically normal throughout the 14-day observation period. One 500 mg/kg bw female rat lost a minimal amount of weight between Days 7 and 14, but overall this animal gained weight during the study. All other animals which survived to scheduled necropsy gained weight during both weeks of the study. The cause of death for the rats which died after treatment with the test substance could not be determined. Treatment-related changes observed at necropsy for the 2000 mg/kg bw female rats which died within 24 hours of dosing consisted of necrosis and hemorrhage in the glandular gastric mucosa. No treatment-related changes were observed at necropsy for the surviving, 2000 mg/kg bw animals or for any animals receiving the lower dose levels of the test substance. The test substance was a gastric irritant as evidenced by necrosis and hemorrhage in the glandular gastric mucosa of rats from the 2000 mg/kg bw dose group. The acute oral LD50 for this test substance was calculated to be greater than 2000 mg/kg bw for male rats and 1414 mg/kg bw for female rats. The combined LD50 (male and female) was determined to be greater than 2000 mg/kg bw.

Acute toxicity inhalation

In an acute inhalation toxicity study conducted according to OECD guideline 403, two groups of five male and five femaleSprague-Dawley rats were exposed to analyticalvapor concentrations of 4.48 (the highest attainable concentration) or 2.50 mg/L of the test substance, as determined by gas chromatography with flame ionization detection, by inhalation for a single four-hour period. The animals were observed for 14 days following the exposure for clinical signs of toxicity. Body weights were measured at least weekly. After 14 days, the animals were euthanatized and necropsied for gross pathology. No mortality occurred during the exposure or during the observation period.Abnormal clinical signs were limited to the high-concentration group and consisted of porphyrin discharges around the nose, red discoloration of the hair, and unkempt hair. All high-concentration animals appeared normal between Days 2 and 14. All low-concentration animals appeared normal throughout the 14 -day observation period. The porphyrin discharges and red discoloration observed for the high-concentration group are suggestive of slight upper respiratory tract irritation (sensory irritation).All animals gained weight during both weeks of the study. No test substance-related changes were detected at necropsy, and no tissues were collected for histological examination. The four-hour LC50 values for this test substance were geater than 4.48 mg/L (highest attainable concentration) for both male and female rats.  

Acute toxicity dermal

An acute dermal toxicity study was conducted according to OECD guideline 402 in male and female rats administered a single limit dose of 2000 mg/kg bwof the test substance topically. All animals appeared normal throughout the 14 -day observation period. No mortality was observed, and all animals gained weight. No treatment-related changes were observed at necropsy, and no tissues were collected for histological examination. The acute dermal LD50 for this test substance was greater than 2000 mg/kg for male and female rats.


Justification for selection of acute toxicity – oral endpoint
Most reliable study

Justification for selection of acute toxicity – inhalation endpoint
Most reliable study

Justification for selection of acute toxicity – dermal endpoint
Most reliable study

Justification for classification or non-classification

Based on the results obtained in the acute oral toxicity study (LD50, female rat = 1414 mg/kg bw) the test item is classified Xn and labelled with R22 "Harmful if swallowed" according to 67/548/EEC (DSD) and classified in acute toxicity category 4 (H302: Harmful if swallowed) according to Regulation (EC) No 1272/2008 (GHS). This classification is done for safety reasons based on the LD50 for females even though the combind LD50 is greater than 2000 mg/kg bw.

Based on availabe studies, the test item is not classified for acute toxicity dermal and inhalation according to the criteria of Directive 67/548/EEC (DSD) and Regulation (EC) No 1272/2008 (CLP).