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Administrative data

Description of key information

Oral: In an acute oral toxicity study to rats the effect of the test item was determined according to OECD guideline 401. The LD50 was determined to be 3600 mg/kg bw. 
Inhalation: The acute inhalation toxicity effect of the test item was determined in a study according to OECD guideline 403, EU method B2 and OPPTS 870.1300. The LC50 was determined to be 10.5 mg/L air.
Dermal: In an acute dermal toxicity study to Wistar rats the effect of the test item was determined according to OECD guideline 402, EU method B3 and OPPTS 870.1200. The LD50 was determined to be >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:
1987-04-07 to 1987-05-06
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study, no GLP study
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: F. Winkelmann, 4799 Borchen
- Fasting period before study: 16 hours before administration
- Housing: 1-5 animals in Makrolon Cages Typ III
- Diet: R10 Alleindiät für Ratten, Ssniff Spezialfutter GmbH, 4770 Soest, ad libitum
- Water: Tap water ad libitum
- Acclimation period: 4-8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 1 °C
- Humidity (%): 60 % +/- 5 %
- Air changes (per hr): 15 times /hour
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 6310 mg/kg

DOSAGE PREPARATION: Rats were fasted for 16 hours before application of the test item; administration volume was 3.024-7.602 cm3/kg of undiluted test material.
Doses:
2510, 3160, 3980, 5010 and 6310 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 and weighing: Weighing after 1, 7 and 14 days.
- Symptoms: After 6 hours of treatment
- Necropsy of survivors performed: yes
- Other examinations performed: mean values of body weight
Statistics:
LD50 values calculated according to Litchfield and Wilcoxon.
Preliminary study:
none
Sex:
male/female
Dose descriptor:
LD50
Effect level:
3 600 mg/kg bw
Based on:
test mat.
Mortality:
2510 mg/kg: 2 animals
3160 mg/kg: 5 animals
3980 mg/kg: 6 animals
5010 mg/kg: 6 animals
6310 mg/kg: 10 animals
Clinical signs:
After 30 min the animals showed symptoms like sedation, ventral and lateral position, ataxia, shiver, stagger, non controlled movements, squat position, decelerate respiration.

Later symptoms were rough fur, hypothermia, bloody urine, glassy eyes, loud breathing, emaciation, bloody rhinorrhoea and aggressive behavior. In the higher dose groups some animals showed morbidity symptoms after 1 hour. 5 days after application all survived animals were without symptoms.
Body weight:
No abnormalities.
Gross pathology:
Autopsy results:
- Liver and spleen damages
- Bloody urin
- Mucosa was hyperemic
- Red colored stomach content
- Corrosion of stomach mucosa
- White colored pancreas
- Renal pelvis hyperemic
- Dark colored kidneys and dried, hard caecum contents.
- One animal showed smaller abdominal organs, overblown stomach and intestine mucosa, exceeding charge of colon and rectum with dry and hard dung and one clut of blood in the bladder.
- Scared liver
- Color changes of kidneys
- Cirrhosis like changement of the medulla of kidneys
- Swellings on the gastro-oesophageal vestibule
- One animal showed a yellow colored, flattened kidney.
- One animal showed a centre of inflammation on the liver.
- In one animal the diaphragm growed together with stomach, pancreas and melt. Between pancreas and melt was a abscess filled out with sanies.
Other findings:
- Dark color change of the tails and the tail tips.
- Concerned tails were necrotic and of soft consistence. They dried out and droped out after 5-10 days. One animal lost almost the complete tail.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
3 600 mg/kg bw
Quality of whole database:
Guideline study, no GLP 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:
2014-03-24 to 2014-05-27
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline and GLP study
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
2009
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.2 (Acute Toxicity (Inhalation))
Version / remarks:
2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1300 (Acute inhalation toxicity)
Version / remarks:
1998
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories, B.V., Kreuzelweg 53, 5961 NM Horst/Netherlands
- Age at study initiation: 10 weeks (group 1), 9 weeks (group 2)
- Weight at study initiation: Males (256.4 to 327 g), Females (174.8 to 206.8 g)
- Housing: Makrolon cages type-4
- Diet: Pelleted standard Harlan Teklad 2914 C rodent maintenance diet, ad libitum
- Water: Community tap-water from Itingen, ad libitum
- Acclimation period: Performed for 15 days (group 1) or 7 days (group 2)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/-3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 10-15 air changes
- Photoperiod (hrs dark / hrs light): 12 hour fluorescent light/12 hour dark cycle with at least eight hours music during the light period.
Route of administration:
inhalation: vapour
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Nose only, flow past exposure chamber
- Exposure chamber volume: Flow of air at each tube was 1.0 L/min.
- Method of holding animals in test chamber: The animals were confined separately in restraint tubes which were positioned radially around the exposure chamber.
- Source and rate of air: Flow of air at each tube was 1.0 L/min.
- Method of conditioning air: The test item was pumped into a glass flask. Compressed, filtered, and dried air was supplied into the glass flask through a metal nebulization tube. The glass flask was kept at a temperature of 28 to 29 °C with a thermal regulating device set to facilitate the process of vaporization.


TEST ATMOSPHERE
- Brief description of analytical method used: Test atmosphere samples were collected at room temperature in a solvent trap constructed with two fritted wash bottles with approximately 100 mL acetonitrile (100%, lot nos. 1325508002 and 142308006, JT Baker) each, connected one after the other to prevent carry-over and ensure all test item was trapped (samples B1 and B2). The wash bottles were placed in a bucket and cooled with an ice/water mixture. All test item was trapped in the first wash bottle (samples B1). The duration of sampling was 10 min. The samples were filled up to 100 g of Acetonitrile and transferred into appropriate labeled vials and forwarded immediately at ambient temperature to the scientist responsible for formulation analysis and stored at -20 ± 5 °C until analysis. The samples were analyzed using a GC method.
- Samples taken from breathing zone: yes
- Particle size distribution: The concentration of the test item in the inhalation chamber was controlled by regulating the flow of the test item into the atmosphere generation glass flasks and by dilution with compressed, filtered, and dried air to achieve the concentrations required for this study.- Rationale for the selection of the starting concentration: The target concentration of 20 mg/L air for 4 hours is the recommended concentration for a limit test (OECD 403, “Acute Inhalation Toxicity”). The target concentration for group 2 was based on regulatory cutoff values.
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
Target vapour concentration: 10 and 20 mg/L air.
Chemically determined mean concentration: 10.5 mg/L air and 22.0 mg/L air.
No. of animals per sex per dose:
10 animals per dose
5 males and 5 females
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: Viability/Mortality: Once daily during the acclimatization period, once before exposure on the day of exposure (test day 1), three times during exposure, immediately and 1 h after exposure on test day 1 and twice daily during the observation period.
Clinical Signs: Once daily during the acclimatization period, once before exposure on the day of exposure (test day 1), three times during exposure at hourly intervals (only grossly abnormal signs were detectable during exposure as animals were restrained in the exposure tubes), immediately and 1 h after exposure on test day 1 and once daily during the observation period.
Body Weights: Once during acclimatization, on test days 1 (before exposure), 2, 4, 8 and 15 (before necropsy).
- Necropsy of survivors performed: yes
Statistics:
No statistical analysis was performed as only two groups were allocated to the study.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 10.5 - < 22 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
All animals of group 1 exposed to a concentration of 22.0 mg/L air died within 3 hours after exposure start. Two males and one female of group 2 exposed to a concentration of 10.5 mg/L air died within 1 to 7 days after exposure. The remaining seven animals survived the scheduled observation period.
Clinical signs:
other: No clinical signs were visible in animals of group 1 exposed to a concentration of 22.0 mg/L air one hour after exposure start. Two hours after exposure start, four males and two females were unconscious. All animals died within 3 hours after exposure sta
Body weight:
All surviving animals of group 2 exposed to 10.5 mg/L air lost body weight during the first week after treatment but had started to gain body weight on test day 8, with the exception of one female, which already started to gain weight on test day 4. Due to its premature death, no body weight was recorded after exposure of male no. 11 which was found dead on test day 2. The male and the female which were found dead on test day 8 and test day 3, respectively (animal nos. 14 and 17, respectively), had continuously lost body weight until their death. In the second week after exposure, all surviving animals showed normal body weight gain.
Gross pathology:
All animals of group 1 which died during exposure to a concentration of 22.0 mg/L air showed dark red discoloration of the lungs at necropsy. No test item-related findings were noted in all females and the surviving males of group 2 exposed to a concentration of 10.5 mg/L air. The male which was found dead on test day 2 showed reddish discoloration of the lungs, the abdominal cavity of the male which was found dead on test day 8 contained watery fluid.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LC50
Value:
10 500 mg/m³
Quality of whole database:
GLP and guideline 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:
2013-06-19 to 2013-09-30
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
1987
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
1998
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: Young adult animals (male animals approx. 8 weeks, female animals approx. 12 weeks)
- Weight at study initiation: Animals of comparable weight (± 20% of the mean weight)
- Housing: Housed in fully air-conditioned rooms.
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany)
- Water: Tap water ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 °C +/- 3 °C
- Humidity (%): 30 - 70 %
- Air changes per hour: Approx. 20

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: about 40 cm2, the clipped epidermis, covering of the application site with a semi-occlusive dressing
- % coverage: at leat 10 % of the body surface

REMOVAL OF TEST SUBSTANCE
- Washing: with warm water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied: Administration volume: 2.41 mL/kg bw
- Concentration: undiluted
Duration of exposure:
24 hours
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: bodyweight: Shortly before administration, weekly thereafter and on the last day of observation
Clinical observations: Several times on the day of administration, and at least once daily thereafter each workday
Scoring of skin findings: 30-60 minutes after removal of the semi-occlusive dressing.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, histopathology.
Statistics:
Calculations were performed using Microsoft Excel 2003 and checked with a calculator.
Preliminary study:
none
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occured.
Clinical signs:
No adverse clinical signs were observed during clinical examination.
Body weight:
No abnormalities occured.
Gross pathology:
No local effects were observed.
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 study

Additional information

Acute toxicity: Oral

In an acute oral toxicity study to rats the effect of the test item was determined according to OECD guideline 401. Concentrations tested were 2510, 3160, 3980, 5010 and 6310 mg/kg. The LD50 was determined to be 3600 mg/kg bw. The animals showed color changes in the liver and the kidneys. Moreover a swelling of the Mucosa of the gastro-oesophagael vestibule and a completely lost of the tail was observed. No abnormalities occured in the body weight.

Acute toxicity: Inhalation

The acute inhalation toxicity effect of the test item was determined in a study according to OECD guideline 403, EU method B2 and OPPTS 870.1300. Two groups of five male and five female albino rats each were exposed by nose-only, flow-past inhalation for four hours to the test item at chemically determined mean concentrations of 10.5 mg/L air and 22.0 mg/L air, respectively. All animals were observed for clinical signs and mortality during the inhalation exposure and the subsequent 14-day observation period. Body weights were recorded at acclimatization start, prior to exposure on test day 1 and during the observation period on test days 2, 4, 8 and 15 before necropsy. On test day 15 all surviving animals were sacrificed and necropsied. The vapor concentration, temperature, relative humidity, oxygen content and airflow rate measured during the exposure were considered to be satisfactory for a study of this type. All animals exposed to 22.0 mg/L air died within 3 hours after exposure start. Two males and one female of group 2 exposed to a concentration of 10.5 mg/L air died within 1 to 7 days after exposure. The remaining seven animals survived the scheduled observation period. Four males and two females exposed to 22.0 mg/L air were unconscious before their death within 3 hours after exposure start. During and after exposure on test day 1, all animals exposed to 10.5 mg/L air showed clinical signs of toxicity affecting behavior, appearance and breathing. Slight tachynea was observed until test day 2 in all animals; slightly ruffled fur persisted up to test day 6. From test day 7 onwards, all animals were free from clinical signs. During the first week after exposure, body weight loss was noted in all animals exposed to 10.5 mg/L air. In the second week after exposure, all surviving animals showed normal body weight gain. All animals which died during exposure to 22.0 mg/L air showed dark red discoloration of the lungs at necropsy. No test item-related findings were noted in all females and the surviving males exposed to a concentration of 10.5 mg/L air. The male which was found dead on test day 2 showed reddish discoloration of the lungs, the abdominal cavity of the male which was found dead on test day 8 contained watery fluid. Based on the findings of this study, the LC50 for 4-hour exposure of the item was considered to be between 10.5 and 22.0 mg/L air (chemically determined mean vapor concentration). There was no indication of relevant sex-related differences in toxicity of the test item.

Acute toxicity: Dermal

In an acute dermal toxicity study to Wistar rats the effect of the test item was determined according to OECD guideline 402, EU method B3 and OPPTS 870.1200. In an acute dermal toxicity study (limit Test), young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 2000 mg/kg bw (undiluted) of the test item to the clipped skin (dorsal and dorso-lateral parts of the trunk) and covered by semi-occlusive dressing for 24 hours. The application area comprised at least 10% of the total body surface area. The animals were observed for 14 days. The following test item-related clinical observations were recorded during the course of the study:

· No signs of systemic toxicity or skin effects were observed in the animals.

· No macroscopic pathologic abnormalities were noted in the animals examined at the end of the study.

· The mean body weight of the animals increased within the normal range throughout the study period. No mortality occurred. Accordingly, the acute dermal median lethal dose (LD50) was determined to be LD50, dermal, rat > 2000 mg/kg bw.

Justification for classification or non-classification

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

The available experimental test data is reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on the result of the acute inhalation study the substance is considered to be classified as acute toxic category 4, H332 under Regulation (EC) No 1272/2008, as amended for the sixth time in Regulation No 605/2014.

The substance is not considered to be classified for acute dermal toxicity and for acute oral toxicity under Regulation (EC) No 1272/2008, as amended for the sixth time in Regulation No 605/2014.