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Administrative data

Description of key information

In an acute oral toxicity study according to OECD 401, the LD50 of the test item was determined to be greater than 5000.0 mg/kg bw in rats.

In an acute inhalation toxicity study according to OECD 403, the LC50 of the test item was determined to be greater than 2120 mg/m3.

In an acute dermal toxicity study according to OECD 402, the LD50 value of the test item was determined to be greater than 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:
December 22, 1994 - January 5, 1995
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
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hiltop Lab Animals, Scottdale
- Weight at study initiation: 197-216 g (males), 201-215 g (females)
- Fasting period before study: approximately 21 hours
- Housing: individually (suspended stainless steel caging with mesh floors)
- Diet: Purina Rodent Chow (Feed was replaced approximately 3 hours after dosing.)
- Water: Tap water (ad libitum)
- Acclimation period: 16 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.6 – 22.2
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 4.71 mL/kg

DOSAGE PREPARATION: Individual doses were calculated based on the initial bodyweights, taking into account the specific gravity of the test substance.

Doses:
5000 mg/kg bw/day
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Individual weights of the animals were recorded just prior to test substance administration (initial) and again on days 7 and 14 (termination). The animals were observed for signs of gross toxicity, behavioral changes and mortality at 0.5, 1, 3 and 20.75 hours post-dosing and at least once daily thereafter for 14 days.
- Necropsy of survivors performed: yes

Preliminary study:
A preliminary rangefinding screen was conducted at dose levels of 100, 300, 1000, 3000 and 5000 mg/kg bw. The test substance was administered by oral gavage to one male and one female per dose level. The test substance was administered to each animal and the animals were observed daily for 7 days. Based on the range finding screen, a dose level of 5000 mg/kg bw was selected for the limit test.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Mortality:
Male: 5000 mg/kg bw; Number of deaths: 0 of 10
Female: 5000 mg/kg bw; Number of deaths: 0 of 10
Clinical signs:
Within thirty minutes of administration, all animals developed clinical signs including hunched posture, piloerection, irregular respiration, hypoactivity, diarrhea and ano-genital staining.

Body weight:
There was no remarkable body weight change.
Gross pathology:
There were no test article related findings noted.
Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results of testing, the single dose Acute Oral Toxicity LD50 of the test substance is greater than 5000 mg/kg of bodyweight when administered as received.
Executive summary:

An Acute Oral Toxicity test according to OECD Guideline 401 was conducted with rats to determine the potential for the test item to produce toxicity via the oral route. A preliminary rangefinding screen was conducted at dose levels of 100, 300, 1,000, 3000 and 5000 milligrams of the test substance per kilogram of bodyweight. The test substance was administered by oral gavage to one male and one female per dose level. Based on the results of the rangefinding screen, 5000 mg/kg bw was selected as the dose level for the full test.

Ten (five males and five females) healthy rats were selected for the full test. Each animal received 5000 mg/kg bw of the test substance by oral intubation using a stainless steel ball-tipped gavage needle attached to an appropriate syringe.

The animals were observed for signs of gross toxicity and mortality at least once daily for 1 4 days. Bodyweights were recorded just prior to administration and again on days 7 and 14 (termination). Necropsies were performed on all animals at terminal sacrifice.

All animals survived and gained weight. Following administration, all rats developed clinical signs including hunched posture, piloerection, irregular respiration, hypoactivity, diarrhea and ano-genital staining. All animals recovered by day 4 and appeared active and healthy for the remainder of the 14-dav observation period. Gross necropsy findings at terminal sacrifice were generally unremarkable.

Based on the results of testing, the single dose Acute Oral Toxicity LD50 of the test substance is greater than 5000 mg/kg of bodyweight when administered as received.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
5 000 mg/kg bw
Quality of whole database:
GLP and Guideline conform 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:
December 15, 1994 - February 3, 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hilltop Lab Animals, Scottdale
- Weight at study initiation: 228-239 g (males), 220-235 g (females)
- Housing: individually (in suspended stainless steel caging with mesh floors)
- Diet: Purina Rodent Chow
- Water: Tap water (ad libitum except during exposure)
- Acclimation period: 23 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.6 – 23.9
- Photoperiod (hrs dark / hrs light): 12/12


Route of administration:
inhalation: aerosol
Type of inhalation exposure:
whole body
Vehicle:
clean air
Mass median aerodynamic diameter (MMAD):
2.2 µm
Geometric standard deviation (GSD):
1.93
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Rectangular whole body perspex chamber with prechamber operated under slight negative pressure
- Exposure chamber volume: 150 L
- Source and rate of air: Approximately 20 liters per minute (Lpm) was supplied from a compressed gas cylinder (Airco #3001) of dry, breathing-grade air to the spray atomization nozzle. Compressed airflow was measured with a Dwyer Rotameter Model #VFB (0-40 Lpm). Approximately 20.6 Lpm of filtered conditioned room air was supplied as diluent air. Room airflow was measured with an Omega Mass Flow Meter, Model #FMA 5613. Chamber airflow was monitored throughout the exposure period and recorded periodically. Total airflow ranged from 40.3 to 40.7 with a mean of 40.6 Lpm,
- System of generating aerosols: The test atmosphere was generated using a 1/4 inch JCO atomizer, FC4 fluid cap and 1502 air cap (Spraying Systems Inc.). Compressed air was supplied at 25 psi. The test substance was metered to the atomization nozzle through Size 14 Master Flex Tygon tubing, using a Master Flex Pump Model 7520-35.
- Method of particle size determination: An eight-stage Andersen cascade impactor was used. Samples were withdrawn from the breathing zone of the animals on two occasions. The filter paper collection stages were weighed before and after sampling to determine the mass collected at each stage. The aerodynamic mass median diameter and geometric standard deviation were determined graphically using two cycle logarithmic probit axes.
- Temperature, humidity, pressure in air chamber: 21.7-23.3°C, 40-45%, 25 psi

TEST ATMOSPHERE
- Brief description of analytical method used: Gravimetric samples were withdrawn on eight occasions from the breathing zone of the animals. Samples were collected using 25 mm glass fiber filters (GF/B Whatman) in filter holders attached by % inch tygon tubing to a General Electric vacuum pump Model 5KH10GGR28. Filter papers were weighed before and after collection to determine the mass collected. This value was divided by the total volume of air sampled to determine the chamber concentration. The collections were carried out for 3 minutes at airflows of 4 Lpm. Sample airflows were measured using an Omega Flow Meter Model #FMA 5610.
- Samples taken from breathing zone: yes


Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
2.12 mg/L (gravimetric chamber concentration)
10.93 mg/L (nominal concentration)
No. of animals per sex per dose:
5 male, 5 female
Control animals:
no
Details on study design:
- Duration of observation period following administration: 50 days
- Frequency of observations and weighing: Individual weights of the animals were recorded just prior to test substance exposure (initial) and again on days 7, 14, 21, 28, 35, 42 and 49 or after death. Cage-Side observations were made at least every 30 minutes during exposure, upon chamber removal and at least once daily thereafter for 50 days.
- Necropsy of survivors performed: yes
- Other examinations performed: histopathology (lung tissue)
Preliminary study:
Prior to initiation of the full inhalation study, pre-test trials were conducted to establish generation procedures for achieving as closely as possible the desired (2.0 mg/L) or highest possible chamber concentration and desired particle size distribution (mass median aerodynamic diameter less than or equal to 4/ µm). The exposure procedures and atomization equipment used were based on results of pre-test trial number 2 which provided a gravimetric concentration of 2.0 mg/L and a mass median aerodynamic diameter of 2.1 µm.
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 2.12 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
One male animal died on test day 11.
Clinical signs:
In chamber animal observation included ocular and nasal discharge, facial staining, irregular respiration, dyspnea, gasping, hunched posture and hypoactivity. Upon chamber removal, similar clinical signs persisted in all animals. Within several days of exposure animals also developed piloerection, rales, emaciation, ano-genital staining and an unthrifty appearance. Prior to death, the decedent exhibited reduced food consumption, reduced fecal volume and abdominal distention. Transient signs of abnormal respiration (rales and irregular respiration! were noted in several rats for an extended period of time.

Body weight:
Transient body weight loss was noted during the study.
Gross pathology:
Gross necropsy of the decedent revealed discoloration of the lungs, gastro intestinal tract and liver, gaseous distention of the gastrointestinal tract, edema of the lungs and retraction of the testes into the abdominal cavity. Gross necropsy findings at terminal sacrifice were generally unremarkable. Apart from red lung discoloration which is consistent with euthanasia by CO2 inhalation, all tissues and organs appeared normal.
Other findings:
- Histopathology: Histological evaluation of the lung tissue of the survivors revealed pulmonary congestion: the presence of alveolar macrophages; the accumulation of mucous in the bronchi and bronchioles; and chronic interstitial inflammation. Although the congestion may have been due to the euthanasia procedure, it along with interstitial inflammation may have resulted from exposure to a very mild irritant.
Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results of testing, the single exposure Acute Inhalation Toxicity LC50 of the test substance is greater than 2.12 mg/L.
Executive summary:

An Acute Inhalation Toxicity test according OECD Guideline 403 was conducted with rats to determine the potential of the test item to produce toxicity via the inhalation route.

After establishing the desired generation procedures during pre-test trials, ten healthy rats were exposed to the test atmosphere for 4 hours. Chamber concentration and particle size distribution of the test substance were determined periodically during the exposure period. The animals were observed for signs of gross toxicity and mortality at least once daily for 50 days. Bodyweights were recorded just prior to exposure and again or days 71 4, 21, 28, 35, 42 and 49 or after death. Gross necropsies were performed on all animals. The lungs of all survivors were removed at terminal sacrifice, preserved in formalin and evaluated histopatholggically.

One male died on day 11 of the study. All other animals survived exposure to the test atmosphere. The gravimetric chamber concentration was 2.12 mg/L. Based on graphic analysis of the particle size distribution as measured with an Anderson Cascade Impactor, the mass median aerodynamic diameter was estimated to be 2.2 µm.

In chamber animal observations included ocular and nasal discharge, facial staining, irregular respiration, dyspnea, gasping, hunched posture and hypoactivity. Upon chamber removal, similar clinical signs persisted in all animals. Within several days of exposure animals also developed piloerection, rales, emaciation, ano-genital staining and an unthrifty appearance. Prior to death, the decedent exhibited reduced food consumption, reduced fecal volume and abdominal distention. Transient signs of abnormal respiration were noted in several rats for an extended period of time. In order to assess the reversal of symptoms, the observation period was extended to 50 days. Although clinical signs persisted and transient bodyweight loss was noted during the study, all surviving rats gained weight over the entire 50 day observation period. Gross necropsy of the decedent revealed discoloration of the lungs, gastro-intestinal tract and liver, gaseous distention of the gastro-intestinal tract, edema of the lungs and retraction of the testes into the abdominal cavity. Gross necropsy findings at terminal sacrifice were generally unremarkable. Apart from red lung discoloration which is consistent with euthanasia by CO2 inhalation, all tissues and organs appeared normal. In order to further investigate the abnormal respiratory findings noted during the observation period, the lungs of all surviving animals were removed at gross necropsy and preserved in neutral buffeted formalin. Histological evaluation of the lung tissue revealed pulmonary congestion: the presence of alveolar macrophages; the accumulation of mucous in the bronchi and bronchioles; and chronic interstitial inflammation. Although the congestion may have been due to the euthanasia procedure, it along with interstitial inflammation may have resulted from exposure to a very mild irritant.

Based on the results of testing, the single exposure Acute Inhalation Toxicity LC50 of the test substance is greater than 2.12 mg/L.

 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating conc.
2 120 mg/m³
Quality of whole database:
GLP and Guideline conform 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:
14 December 1994 - 28 December 1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hilltop Lab Animals, Scottdale, PA
- Weight at study initiation: 235-265 g (males), 210-232 g (females)
- Housing: individually in suspended stainless steel caging with mesh floors
- Diet: Pelleted Purina Rodent Chow #5012
- Water: ad libitum (tap water)
- Acclimation period: 22 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.6 - 22.2
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: 2x3"
- % coverage: 10% of the body surface
- Type of wrap if used: 2 1/4 x 3" adhesive backed-gauze patch, which was wrapped with 3" Durapore tape

REMOVAL OF TEST SUBSTANCE
- Washing: Yes
- Time after start of exposure: 24 hours

Duration of exposure:
24 h
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 males, 5 females
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Body weights were recorded just prior to test substance application (initial) and again on days 7 and 14 (termination). Cage-side observations were done at 1 and 24 hours after application and at least once daily thereafter for 14 days.
- Necropsy of survivors performed: yes
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no deaths during the study.
Clinical signs:
One female animale exhibited alopecia.
Body weight:
No test article related findings were observed.
Gross pathology:
No findings for any examined tissues and/or organs were noted.
Other findings:
- Other observations: Dermal irritation was noted at the administration site of
all animals.
Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results of testing, the single dose Acute Dermal Toxicity LD50 of the test substance is greater than 2000 mg/kg of bodyweight when applied as received.
Executive summary:

An Acute Dermal Toxicity test was conducted with rats to determine the potential for the test item to produce toxicity after topical application.Two thousand milligrams of the test substance per kilogram of bodyweight was applied to the skin of ten healthy rats for 24 hours.The animals were observed for signs of gross toxicity and mortality at least once daily for 14 days. Bodyweights were recorded just prior to application and again on days 7 and 14 (termination). Necropsies were performed on all animals al terminal sacrifice.

All animals survived and gained weight. One female exhibited alopecia from day 4 through 9. All other animals appeared active and healthy. Apart from the dermal irritation noted at the dose site of all animals, there were no other signs of gross toxicity, adverse pharmacologic effects or abnormal behavior. Gross necropsy findings at terminal sacrifice were generally unremarkable.

Based on the results of testing, the single dose Acute Dermal Toxicity LD50 of the test substance is greater than 2000 mg/kg of bodyweight when applied as received.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
2 000 mg/kg bw
Quality of whole database:
GLP and Guideline conform study

Additional information

Acute oral toxicity

An Acute Oral Toxicity test according to OECD Guideline 401 was conducted with rats to determine the potential for the test item to produce toxicity via the oral route. A preliminary rangefinding screen was conducted at dose levels of 100, 300, 1,000, 3000 and 5000 milligrams of the test substance per kilogram of bodyweight. The test substance was administered by oral gavage to one male and one female per dose level. Based on the results of the rangefinding screen, 5000 mg/kg bw was selected as the dose level for the full test.

Ten (five males and five females) healthy rats were selected for the full test. Each animal received 5000 mg/kg bw of the test substance by oral intubation using a stainless steel ball-tipped gavage needle attached to an appropriate syringe.

The animals were observed for signs of gross toxicity and mortality at least once daily for 1 4 days. Bodyweights were recorded just prior to administration and again on days 7 and 14 (termination). Necropsies were performed on all animals at terminal sacrifice.

All animals survived and gained weight. Following administration, all rats developed clinical signs including hunched posture, piloerection, irregular respiration, hypoactivity, diarrhea and ano-genital staining. All animals recovered by day 4 and appeared active and healthy for the remainder of the 14-dav observation period. Gross necropsy findings at terminal sacrifice were generally unremarkable.

Based on the results of testing, the single dose Acute Oral Toxicity LD50 of the test substance is greater than 5000 mg/kg of bodyweight when administered as received.

Acute inhalation toxicity

An Acute Inhalation Toxicity test according OECD Guideline 403 was conducted with rats to determine the potential of the test item to produce toxicity via the inhalation route.

After establishing the desired generation procedures during pre-test trials, ten healthy rats were exposed to the test atmosphere for 4 hours. Chamber concentration and particle size distribution of the test substance were determined periodically during the exposure period. The animals were observed for signs of gross toxicity and mortality at least once daily for 50 days. Bodyweights were recorded just prior to exposure and again or days 71 4, 21, 28, 35, 42 and 49 or after death. Gross necropsies were performed on all animals. The lungs of all survivors were removed at terminal sacrifice, preserved in formalin and evaluated histopatholggically.

One male died on day 11 of the study. All other animals survived exposure to the test atmosphere. The gravimetric chamber concentration was 2.12 mg/L. Based on graphic analysis of the particle size distribution as measured with an Anderson Cascade Impactor, the mass median aerodynamic diameter was estimated to be 2.2 µm.

In chamber animal observations included ocular and nasal discharge, facial staining, irregular respiration, dyspnea, gasping, hunched posture and hypoactivity. Upon chamber removal, similar clinical signs persisted in all animals. Within several days of exposure animals also developed piloerection, rales, emaciation, ano-genital staining and an unthrifty appearance. Prior to death, the decedent exhibited reduced food consumption, reduced fecal volume and abdominal distention. Transient signs of abnormal respiration were noted in several rats for an extended period of time. In order to assess the reversal of symptoms, the observation period was extended to 50 days. Although clinical signs persisted and transient bodyweight loss was noted during the study, all surviving rats gained weight over the entire 50 day observation period. Gross necropsy of the decedent revealed discoloration of the lungs, gastro-intestinal tract and liver, gaseous distention of the gastro-intestinal tract, edema of the lungs and retraction of the testes into the abdominal cavity. Gross necropsy findings at terminal sacrifice were generally unremarkable. Apart from red lung discoloration which is consistent with euthanasia by CO2 inhalation, all tissues and organs appeared normal. In order to further investigate the abnormal respiratory findings noted during the observation period, the lungs of all surviving animals were removed at gross necropsy and preserved in neutral buffeted formalin. Histological evaluation of the lung tissue revealed pulmonary congestion: the presence of alveolar macrophages; the accumulation of mucous in the bronchi and bronchioles; and chronic interstitial inflammation. Although the congestion may have been due to the euthanasia procedure, it along with interstitial inflammation may have resulted from exposure to a very mild irritant.

Based on the results of testing, the single exposure Acute Inhalation Toxicity LC50 of the test substance is greater than 2.12 mg/L.

Acute dermal toxicity

An Acute Dermal Toxicity test was conducted with rats to determine the potential for the test item to produce toxicity after topical application.Two thousand milligrams of the test substance per kilogram of bodyweight was applied to the skin of ten healthy rats for 24 hours.The animals were observed for signs of gross toxicity and mortality at least once daily for 14 days. Bodyweights were recorded just prior to application and again on days 7 and 14 (termination). Necropsies were performed on all animals at terminal sacrifice.

All animals survived and gained weight. One female exhibited alopecia from day 4 through 9. All other animals appeared active and healthy. Apart from the dermal irritation noted at the dose site of all animals, there were no other signs of gross toxicity, adverse pharmacologic effects or abnormal behavior. Gross necropsy findings at terminal sacrifice were generally unremarkable.

Based on the results of testing, the single dose Acute Dermal Toxicity LD50 of the test substance is greater than 2000 mg/kg of bodyweight when applied as received.

 

 

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

Based on the available experimental data, the test item is not considered to be classified for acute oral, inhalation and dermal toxicity under Regulation (EC) No 1272/2008, as amended for the ninth time in Regulation (EC) No 2016/1179.