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Administrative data

Description of key information

Oral: OECD 425. LD50, rat.  The LD50 was 1750 mg/kg.  Reliability = 2 
Dermal: OECD 402. LD50, rat. The LD50 was >5000 mg/kg. Reliability = 1
Inhalation: OECD 403. 4-hr LC50, rat. The LC50 was >5200 mg/m3. Reliability =1.

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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: This study was selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labeling and/or risk assessment.
Qualifier:
according to
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Deviations:
no
Remarks:
The study was conducted according to guideline in effect at time of study conduct.
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Remarks:
The study was conducted according to guideline in effect at time of study conduct.
GLP compliance:
yes
Test type:
up-and-down procedure
Limit test:
no
Species:
rat
Strain:
other: Crl:CD(SD)
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 9-11 weeks
- Weight at study initiation: 267.8-331.2 grams
- Fasting period before study: 16-18 hours
- Housing:All animals were housed singly in stainless steel, wire-mesh cages suspended above cage boards.
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: at least 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-26
- Humidity (%): 30-70
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Doses:
175, 550, 1750, 5000 mg/kg
No. of animals per sex per dose:
175 mg/kg (1 male rat), 550 mg/kg (2 male rats), 1750 mg/kg (4 male rats), 5000 mg/kg (3 male rats)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: The rat were observed for clinical signs at the beginning of fasting, just before dosing (test day 0), once during the first 30 minutes after dosing and 2 more times on the day of dosing, and once each day thereafter.
-Frequency of weighing: The rats were weighed on test days –1, 0, 7, and 14.
- Necropsy of survivors performed: yes
Statistics:
A software package (A0T425StatPgm)a was used to determine the dose progression and the LD50.
Sex:
male
Dose descriptor:
LD50
Effect level:
1 750 mg/kg bw
95% CL:
1 239 - 4 450
Mortality:
Death occurred in one rat dosed at 1750 mg/kg and in all 3 rats dosed at 5000 mg/kg.
Clinical signs:
Lethargy was observed on the day of dosing in the rat dosed at 175 mg/kg. The 2 rats dosed at 550 mg/kg exhibited wet fur up to the day after dosing. Three surviving rats dosed at 1750 mg/kg exhibited wet fur and stained fur/skin up to the day after dosing. One of these rats also exhibited low posture on the day of dosing. The remaining rat dosed at 1750 mg/kg exhibited lethargy, stained fur/skin, and wet fur on the day of dosing and was found dead on the day after dosing. Lethargy was observed on the day of dosing in one of the rats dosed at 5000 mg/kg. This rat was found dead on the day of dosing. Another rat dosed at 5000 mg/kg exhibited lung noise, lethargy, decreased muscle tone, stained fur/skin, and wet fur on the day of dosing and was found dead on the day after dosing. The remaining rat dosed at 5000 mg/kg exhibited lethargy and was found dead on the day of dosing.
Body weight:
No body weight loss occurred in the surviving rats after dosing.
Gross pathology:
Gross findings were present in 1 rat dosed at 1750 mg/kg and 3 rats dosed at 5000 mg/kg. These included skin stain, lungs expanded, eye discoloration, and/or stomach discoloration. No other gross findings were observed.
Interpretation of results:
harmful
Remarks:
Migrated information Criteria used for interpretation of results: other: Directive 67/548/EEC
Conclusions:
The oral LD50 for the test substance was 1750 mg/kg in male rats.
This study and the conclusions which are drawn from it fulfill the quality criteria (validity, reliability, repeatability).
Executive summary:

A single dose of the test substance was administered by oral gavage to 1 fasted male rat at a dose of 175 mg/kg, to 2 fasted male rats at a dose of 550 mg/kg, to 4 fasted male rats at a dose of 1750 mg/kg, and to 3 fasted male rats at a dose of 5000 mg/kg. The rats were dosed one at a time at a minimum of 48-hour intervals. The rats were observed for mortality, body weight effects, and clinical signs for up to 14 days after dosing. All rats were necropsied to detect grossly observable evidence of organ or tissue damage. One of the rats dosed at 1750 mg/kg was found dead on the day after dosing, and the 3 rats dosed at 5000 mg/kg were found dead on the day of dosing or the day after dosing. Clinical signs of toxicity were observed in all rats up to the day after dosing and included lethargy, wet fur, stained fur/skin, lung noise, decreased muscle tone, and/or low posture. No body weight losses occurred in the surviving rats. Gross findings were present in 1 rat dosed at 1750 mg/kg and 3 rats dosed at 5000 mg/kg. These included skin stain, lungs expanded, eye discoloration, and/or stomach discoloration. No other gross findings were observed. Under the conditions of this study, the oral LD50 for the test substance was 1750 mg/kg for male rats. The 95% profile likelihood confidence interval is 1239 to 4450 mg/kg.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
1 750 mg/kg bw

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study was selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labeling and/or risk assessment.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
Remarks:
The study was conducted according to the guideline in effect dated 1981.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
other: Crl:CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: approximately 8 weeks old;
- Weight at study initiation: male rats 254 - 314 grams; female rats 196 - 223 grams
- Fasting period before study: not reported
- Housing: individually in solid bottom caging with bedding
- Diet (e.g. ad libitum): ad libitum, except during exposure
- Water (e.g. ad libitum): ad libitum, except during exposure
- Acclimation period: 6 days prior to testing

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-26ºC
- Humidity (%): 30-70%
- Air changes (per hr): Not reported
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: unchanged (no vehicle)
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Glass (cylindrical) chamber with a baffle inside the chamber to promote uniform chamber distribution of the test atmosphere
- Exposure chamber volume: Nominal internal volume of 34 L
- Method of holding animals in test chamber: Individually restrained in perforated stainless steel cylinders with conical nose pieces, that were inserted into a polymethylmethacrylate faceplate attached to the exposure chamber so that the nose of each animal extended into the exposure chamber.
- Source and rate of air: High-pressure air, metered into the nebulizer by a mass flow controller, carried the resulting atmosphere into the exposure chamber at 13 mg/m³ chamber aerosol concentration. Chamber airflow was at least 10 air changes/hour.
- Method of conditioning air: Not reported
- System of generating particulates/aerosols: Atmospheres were generated by nebulizer; the test substance was metered into the nebulizer with a syringe infusion pump.
- Method of particle size determination: Cyclone preseparator/cascade impactor and constant flow air sampler
- Treatment of exhaust air: A scrubber with water followed by an MSA charcoal/HEPA filter cartridge prior to discharge into the fume hood
- Temperature, humidity, pressure in air chamber: 20-24°C, 30-70%, not reported

TEST ATMOSPHERE
- Brief description of analytical method used: gravimetric analysis at least 4 times in the test chambers and once during the air-only exposure.
- Samples taken from breathing zone: yes

Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
13, 100 and 5200 mg/m³
No. of animals per sex per dose:
low and middle doses had 3 per sex and high dose group had 5 per sex
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 2 - 14 days
- Frequency of observations: mortality and response to alerting stimuli 3 times during each exposure and observed for mortality and clinical signs of toxicity immediately after they were removed from the restrainers following exposure. During the recovery period, all rats were observed each day for mortality.
- Frequency of weighing: weighed and observed for clinical signs of toxicity on the day following exposure and at least twice more during the recovery period
- Necropsy of survivors performed: yes
- Other examinations performed: Respiratory tract tissues (lung, larynx/pharynx, trachea, and nose) from the 0, 13, and 100 mg/m³ groups were evaluated microscopically. There were no microscopic examinations of rats in the 5200 mg/m³ exposure group.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5 200 mg/m³ air (analytical)
Exp. duration:
4 h
Remarks on result:
other: highest concentration tested. Mass median aerodynamic diameter (MMAD) of particles had a geometric mean of 1.7-2.7 µm.
Mortality:
none
Clinical signs:
There were no clinical signs of toxicity observed during any exposure in this study. Immediately following the 100 mg/m³ exposure, all rats displayed red nasal discharge. Red discharge around the eyes, nose and mouth were observed in all rats immediately after the 5200 mg/m³ exposure. Rats from the 5200 mg/m³ exposure group also displayed red stained faces and/or heads for up to 2 days post exposure.
Body weight:
Some minor body weight losses were observed in male and female rats in the 0, 13 and 100 mg/m³ exposure groups. Since the body weight losses in the 13 and 100 mg/m³ exposure groups were similar to that of the control rats, these body weight losses are not considered toxicologically significant. Rats in the 5200 mg/m³ exposure group lost from 2.5 to 6.8% of their original body weight for one or 2 days post exposure.
Gross pathology:
There were no gross observations noted during necropsy.
Other findings:
- Histopathology: There were no test substance-related microscopic findings ( lung, larynx/pharynx, trachea, and nose) in rats in the 0, 13 and 100 mg/m³ exposure groups. No microscopic examination was performed at 5200 mg/m³.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
LC50 > 5200 mg/m³
This study and the conclusions which are drawn from it fulfill the quality criteria (validity, reliability, repeatability).
Executive summary:

One group of 5 male and 5 female rats was exposed to an aerosol atmosphere containing 5200 mg/m³ of the test substance in air to determine the inhalation median lethal concentration (LC50). Two groups of 3 male and 3 female rats each were exposed to 13 and 100 mg/m³ the test substance in air to evaluate respiratory tract pathology. As a control for the pathology evaluation, an additional group of one male and one female rat was exposed to air only. All rats were exposed nose-only for a single 4-hour period. Aerosol atmospheres were generated by nebulization, and concentrations of the test substance were measured by gravimetric analysis. The ammonia vapor concentration was monitored with Draeger tubes. The ammonia concentration measured during the 0 and 13 mg/m³ exposures was less than 1 ppm. During the 100 and 5200 mg/m³ exposures the ammonia concentrations were 21 ppm and 960 ppm, respectively. Rats in the 0, 13, and 100 mg/m³ exposure groups were weighed and observed for clinical signs of toxicity during a 2-day recovery period. The rats in the 5200 mg/m³ exposure group were weighed and observed for clinical signs of toxicity during a 14-day recovery period. Gross examinations were performed on all rats, and respiratory tract tissues (lung, larynx/pharynx, trachea, and nose) from the 0, 13, and 100 mg/m³ groups were evaluated microscopically. There were no microscopic examinations of rats in the 5200 mg/m³ exposure group. No deaths occurred in any exposure group. There were no clinical signs of toxicity observed during any exposure in this study. Immediately following the 100 mg/m³ exposure, all rats displayed red nasal discharge. Red discharges around the eyes, nose and mouth were observed in all rats immediately after the 5200 mg/m³ exposure. Rats from the 5200 mg/m³ exposure group also displayed red-stained faces and/or heads for up to 2 days post exposure. Other than the stains and discharges noted immediately after the 100 and 5200 mg/m³ exposures, there were no other toxicologically significant clinical signs, gross pathological or microscopic findings in any rats from any exposure group. Body weight patterns in the 13 and 100 mg/m³ exposure groups were similar to those exhibited by the control rats. Rats in the 5200 mg/m³ exposure group lost from 2.5 to 6.8% of their original body weight for one or 2 days post exposure, followed by normal weight gain throughout the remainder of the recovery period. Under the conditions of this study, the 4-hour inhalation median lethal concentration (LC50) for aerosols of the test substance in male and female rats was greater than 5200 mg/m³. No microscopic changes were observed in the upper or lower respiratory tract at concentrations up to 100 mg/m³, the highest concentration evaluated microscopically.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
5 200 mg/m³

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study was selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labeling and/or risk assessment.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Remarks:
The study was conducted according to the test guidelines in effect at the time of study conduct.
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
Remarks:
The study was conducted according to the guideline in effect at the time of study conduct.
Qualifier:
according to
Guideline:
other: EEC, Method B.3 Directive 92/69/EEC:
Deviations:
no
Remarks:
The study was conducted according to the guideline in effect at the time of study conduct.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: Crl:CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: Approximately 9 weeks old (males); approximately 10 weeks old (females).
- Weight at study initiation: 295.7 grams (average, males); 236.0 (average, females).
- Fasting period before study: No
- Housing: Singly in stainless steel, wire-mesh cages suspended above cage boards.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 6 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-26°C
- Humidity (%): 30-70%
- Air changes (per hr): Not reported
- Photoperiod (hrs dark / hrs light): An approximate 12-hour light/dark cycle
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: Approximately 37 square centimeters on the dorsal skin.
- % coverage: Thirty-seven square centimeters is equal to approximately 10 percent of the total body surface area of rats in the 200 - 300 g body weight range.
- Type of wrap if used: The test substance was covered with a 2-ply gauze patch. The rats were then wrapped with stretch gauze bandage and self-adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Excess test substance was washed from the dorsal skin of each rat with warm water, and the skin was dried.
- Time after start of exposure: Approximately 24 hours.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5000 mg/kg.
- Concentration (if solution): Undiluted
Duration of exposure:
24 hours
Doses:
5000 mg/kg
No. of animals per sex per dose:
5 males; 5 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: Observations for mortality and signs of illness, injury, and abnormal behavior were made daily throughout the study. Observations for for clinical signs of toxicity and dermal response were made following test substance removal and daily throughout the study (weekends excluded for dermal irritation).
- Frequency of weighing: Prior to treatment (test day 0) and on test days 7 and 14.
- Necropsy of survivors performed: Yes, examinations to detect grossly observable evidence of organ or tissue damage.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Remarks on result:
other: No mortality or clinical signs of systemic toxicity or body weight loss. Erythema in all females the day after application, reversible by 2 days after application. Hyperkeratosis and ulceration cleared by 13 days after application. No gross lesions.
Mortality:
No mortality was observed.
Clinical signs:
No clinical signs of toxicity were observed during the study. Four rats exhibited wet fur (perineum, inguen) and yellow-stained fur/skin (perineum, inguen), after test substance removal. These clinical signs are commonly seen in wrapped rats and, therefore, were not considered test substance related. High posture observed in a rat on test day 4 was not considered test substance related because it was only observed in a single animal. Hair loss observed in 1 rat was considered incidental.
Body weight:
The rats exhibited no body weight losses.
Gross pathology:
No gross lesions were present in the rats at necropsy.
Other findings:
Skin responses:
No erythema or edema was observed on the test site of male rats. All female rats exhibited erythema (score of 2) but no edema on the test site the day after application of the test substance. No erythema was observed in these rats by 2 days after application. Hyperkeratosis was observed on the test site of 8 rats, and ulceration was observed on the test site of 3 rats during the study. All dermal effects cleared by 13 days after application.
Interpretation of results:
other: Classification not required
Remarks:
Criteria used for interpretation of results: other: EEC
Conclusions:
LD50 > 5000 mg/kg
This study and the conclusions which are drawn from it fulfill the quality criteria (validity, reliability, repeatability).
Executive summary:

A single dose of the test substance was applied to the shaved, intact skin of 5 male and 5 female rats at a dose of 5000 mg/kg of body weight. The application site was covered with a semi-occlusive dressing for 24 hours, after which the test substance was removed. The rats were observed for 14 days following application. The rats were necropsied to detect grossly observable evidence of organ or tissue damage at the end of the 15-day test period. No deaths occurred. The rats exhibited no clinical signs of systemic toxicity or body weight loss. No erythema or edema was observed on the test site of male rats. All female rats exhibited erythema (score of 2) but no edema on the test site the day after application of the test substance. No erythema was observed in these rats by 2 days after application. Hyperkeratosis was observed on the test site of 8 rats, and ulceration was observed on the test site of 3 rats during the study. All dermal effects cleared by 13 days after application. No gross lesions were present in the rats at necropsy. Under the conditions of this study, the skin absorption LD50 for the test substance was greater than 5000 mg/kg of body weight when applied to the skin of male and female rats for 24 hours.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
5 000 mg/kg bw

Additional information

Toxicity Description: The substance was evaluated for acute toxicity (lethality) via the acute oral, dermal and inhalation routes of exposure. The oral LD50 values were 1750 and 3129 mg/kg in male and female rats, respectively. In the rat acute oral toxicity study, clinical signs of toxicity were observed in all rats up to the day after dosing and included lethargy, wet fur, stained fur/skin, lung noise, decreased muscle tone, and/or low posture. The oral LD50 was 1030 mg/kg in female mice, however mice have been shown to be kinetically distinct (see toxicokinetics section) from rats and monkeys. Therefore, rat data will be selected over mouse data. In a rat acute dermal toxicity study, male and female rats were exposed to the test substance for 24 hours.  No deaths occurred. The rats exhibited no clinical signs of systemic toxicity or body weight loss. No gross lesions were present in the rats at necropsy. The male and female rat dermal LD50 was greater than 5000 mg/kg. In a rat 4-hour acute inhalation study, no deaths occurred in any exposure group. There were no clinical signs of toxicity observed during any exposure in this study. Immediately following the 100 mg/m³ exposure, all rats displayed red nasal discharge. Red discharges around the eyes, nose and mouth were observed in all rats immediately after the 5200 mg/m³ exposure. Rats from the 5200 mg/m³ exposure group also displayed red-stained faces and/or heads for up to 2 days post exposure. Other than the stains and discharges noted immediately after the 100 and 5200 mg/m³ exposures, there were no other toxicologically significant clinical signs, gross pathological or microscopic findings in any rats from any exposure group. The male and female rat 4-hour LC50 was greater than 5200 mg/m3, the highest concentration tested.


Justification for selection of acute toxicity – oral endpoint
OECD Guideline, GLP study

Justification for selection of acute toxicity – inhalation endpoint
Scientifically valid (similar to OECD Guideline), GLP study

Justification for selection of acute toxicity – dermal endpoint
OECD Guideline, GLP study

Justification for classification or non-classification

Based on the male rat oral LD50 of 1750 mg/kg, the test substance is classified as Cat 4 (harmful if swallowed) for acute oral, according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) regulation (EC) no. 1272/2008. 

Based on the inhalation 4-hour LC50 in rats of >5200 mg/m3 and dermal LD50 in rats of >5000 mg/kg, no classification is required for acute inhalation or dermal endpoints.