Registration Dossier

Administrative data

Description of key information

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 March 01, 1990 to March 21, 1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: E. E. C directive 84/449
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Specific details on test material used for the study:
Purity: 99.3 %
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Species, strain: Ico rat OFA.SD. (IOPS Caw),
Supplier: Iffa-Credo (69592 L'Arbresle Cedex - France),
Age: young adults between 5 and 7 weeks old.
Weight at the start of treatment (main study): from 123 g to 173 g (the individual weighs for each sex varied by no more than 20 % of the mean weight of the animals).

Environmental conditions:
Cages housed by sex and in groups of 5·(or 2 for the preliminary study), in type FI polycarbonate cages (interior dimensions 305 x 180 x 184 mm) for the preliminary study and in type MI (interior dimensions 365 x 225 x 180 mm) for the main study.
Air changes: at least 10 per hour
Temperature: 22 ± 3 °C
Humidity: 30 to 70 %
Lighting: artificial, 12 hours out of 24.
Hygiene: bedding changed once a week; cages changed for each study.
Diet: complete pelleted rat-mouse maintenance diet ad libitum (U.A.R., formula A.04 CR - U.A.R.).
Water: softened and filtered (0.6 µm) mains drinking water ad libitum. Bacteriological and chemical analyses checked twice a year.
Acclimatisation period: 8 days before the start of treatment.
Clinical examinations: at reception then before the start of treatment to ensure that only healthy animals are included in the study.
Identification of animals: perforation of ear pinna before the start of treatment.
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
Single oral administration by gestric gavage - round –ended curved stainless-steel oesophageal probe, 3 mm diameter, Perfektum.
No. of animals per sex per dose:
Preliminary study: 6 males, 6 non-pregnant females;
Main study:
control group - 5 males, 5 non-pregnant females;
groups 2 to 6 (treated) - 25 males, 25 non-pregnant females.
Control animals:
yes
Details on study design:
Preliminary test:
3 groups each composed of 2 males and 2 females were treated, at dose levels of 504, 1008 and 2004 respectively at a volume of 0.42, 0.84 and 1.67 ml/kg of test substance as supplied. The test substance was administered after about 19 h of a water regime and received food 4 h after intubation.

Main test:
Five groups of 5 females and 5 males were dosed with at the levels of 360, 456, 564, 708 and 900 mg/kg, corresponding to
0.30, 0.38, 0.47, 0.59 and 0.75 ml/kg respectively. The control group was treated with the distilled water under the same conditions. The test article was administered once only after about 17 h and 30 min of a water regime and received food 4 h after intubation.

Following examinations were performed:
- Clinical examinations: 15 minutes after intubation, then at 1, 2 and 4 hours and then daily for 14 days.
The daily observations performed, amongst others, included changes in the skin and fur, the eyes, mucous membranes, respiratory system, circulatory system, autonomic and central nervous systems, as well as somato-motor activity and behaviour. Shivering, convulsions, salivation, diarrhoea, lethargy, sleeping and coma were noted with particular attention.
- Observation of body weight: Day -1, Days 1 (before administration of the test article), 8 and 15, as well as at the time of death from Day 2 onwards.
- Necropsy examination of all animals. The abdominal and thoracic cavities were opened and particular attention was paid to the following organs: liver, heart, kidneys and lungs.
Preliminary study:
100 % mortality in all dose groups.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
730 mg/kg bw
Based on:
test mat.
95% CL:
> 651 - < 819
Mortality:
10 % mortality in 564 mg/kg dose group, 50 % in 708 mg/kg dose group, 80 % in 900 mg/kg dose group.
Clinical signs:
At at 360 mg/kg and at 456 mg/kg all animals showed subdued behaviour 15 min, 1 and 2 h after administration, some after 4 h and Day 2.
At 360 mg/kg: All rats were normal on the day following treatment. At 456 mg/kg: All rats were normal on Day 3.
at 564 mg/kg: all animals showed subdued behaviour 15 min to 2 h after treatment. Subdued behaviour or prostration (1 case) was noted in all the animals at 4 h as well as slight red coloured urine in 2 rats. In some cases subdued behaviour was noted on Days 2 - 3 - 8 and 9. All surviving rats were normal on Day 11.
at 708 mg/kg: all animals showed subdued behaviour or clonic convulsions (1 case) at 15 min, subdued behaviour or prostration (2 cases) or clonic convulsions (1 case) at 1 h and subdued behaviour or prostration (3 cases) as well as slight red coloured urine in 3 rats at 2 hours. Subdued behaviour was noted in all the surviving animals 4 h after administration and on Day 2. as well as slight red coloured urine in 2 rats at 4 h. In some cases subdued behaviour was noted on Days 3 - 10 and 13, associated with tremors in 1 rat on Day 3. All surviving animals were normal on Day 3.
at 900 mg/kg: prostration or clonic convulsions (2 cases) were noted in all the animals 15 min after administration then subdued behaviour or prostration was observed in the surviving animals from 1 h to Day 4 as well as slight red coloured urine in 1 rat 2 to 4 h after treatment. Prostration 1 case) was noted on Days 5 and 6. All survivng rats were normal on Day 7.
Body weight:
A difference in the mean weights, was only noted in the males of group 2 (Day 8) and of group 3 (Days 8 – 15) than in the control males. The mortality rate noted in the other groups did not allow analysis of their body weight changes.
Gross pathology:
Some animals that died during the study showed congested areas in the mucosa of the non-glandular stomach and a stenosis of the oesophagus with whitish substance. This oesophagial abnormality was also noted in some rats sacrified at the end of the study (groups 2 - 3 - 4 and 5); the other animals do not present any macroscopic abnormality.
Interpretation of results:
Category 3 based on GHS criteria
Conclusions:
Under the study conditions, the LD 50 of the test substance in the rat (male and female) was determined to be 730 mg/kg bw (651 - 819 mg/kg) by Bliss method and 735 mg/kg bw (641 – 844) by Litchfield & Wilcoxon's method.
Executive summary:

A study was conducted to determine the potential toxic effect of the test substance when administered as a single oral dose to Wistar rats according to OECD Guideline 401. The test substance was administered once only, as supplied and at the dose levels of 360, 456, 564, 708 and 900 mg/kg bw, by the oral route (gastric gavage), in the Sprague-Dawley rat (5 males and 5 females per group). This study was performed in comparison with a control group of 10 rats of both sexes treated with the distilled water under the same conditions. Mortality and abnormal clinical signs were noted 15 min after intubation, then at 1, 2 and 4 h, and then daily for the 14 d study period. All the animals were weighed the day before treatment (Day-1), immediately before administration of the test substance (Day-1), on Days 8 and 15, as well as at time of death from Day-2 onwards. A necropsy was performed for all the animals dead during the study and for all surviving rats after the 14 d study period and the final observation (Day-15). Under the study conditions, the LD 50 of the test substance in the rat (male and female) was determined to be 730 mg/kg bw (651 - 819 mg/kg) by Bliss method and 735 mg/kg bw (641 – 844) by the Litchfield & Wilcoxon’s method (Lheritier, 1990).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
730 mg/kg bw

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
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:
other: EEC directive 84/449/EEC
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
Appearance: pink-coloured cream
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Strain: Crl.: (WI) BR - Wistar, white
Source: Firma Charles River Wiga, Sandhofer Weg 7, 8714 Sulzfeld
Animal identification: coloured marking; cage labelled with the following information: dosage, sex, date of study
Weight range at study initiation: m: 177 - 186 g, f: 168 - 201 g
Housing: collective housing up to a maximum of 5 animals per cage (Macrolon type III)
Illumination: artificial lighting (120 lux) from 7. 00 a.m. - 7.00 p.m.
Temp.: 20 ± 2 °C
Relative humidity: 50- 85 %
Measurement: with thermohygrometer twice daily

Prior to study initiation, the animals were acclimated to laboratory conditions for at least 7 days. 24 h before treatment, the fur was removed with electric clippers from an area of roughly 5 x 10 cm on the back of each animal. The skin was subsequently examined for abrasions and animals with healthy, intact skin were then coloured for individual identification.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
Preparation and application of the test substance:
The test substance was administered as a 66.6 % dilution at 3 ml/kg body weight. The pH value has been adjusted to 6.8 with 25 % ammonic solution. The relative density of the applied solution was 1.002 kg/l. For practical reasons the terms ml and g are considered as identical units.
A single dermal application of the test substance was performed. The substance was held in contact with the skin with a porous gauze dressing and Elastoplast (Beiersdorf).
Duration of exposure:
The exposure period was 24 h.
Doses:
66.6 % dilution at 3 ml/kg body weight
No. of animals per sex per dose:
f males and 5 females
Control animals:
not required
Details on study design:
Range finding
A preliminary range finding test with a dose of 2000 mg/kg bw was conducted on two female rats.

Main study
Clinical observations:
In each animal a number of clinical-toxicological signs were evaluated according to a modified Irwin-Screening procedure (Screening Methods in Pharmacology, R. A. Turner, 1965, p. 26). Any change from the normal condition was noted (increase or decrease) and the degree of severity of any clinical symptoms was assessed. The animals were examined at the following intervals after patch removal: 10 min, 1 h, 2 h, 4 h, 24 h, and thereafter once daily up to day 14.

Skin reactions:
After patch removal, dermal irritation was evaluated once daily for 14 days according to a scheme based on Draize.

Body weights:
The body weights of all animals were recorded immediately before treatment (day 0) and surviving animals were reweighed on days 7 and 14 (termination).

Necropsy:
Animals found dead or killed in extremis were immediately necropsied. The surviving animals were sacrificed by CO2 asphyxiation after 14 days and gross pathological examinations were subsequently performed.

Evaluation of the data:
LD50 values were calculated according to Finney D.Y., Probit Analysis, 3rd edition, Cambridge, 1971.
Preliminary study:
There were no deaths in the preliminary study.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
no indication of skin irritation up to the relevant limit dose level
Mortality:
No animals died during the course of the main study.
Clinical signs:
Some animals showed a slight to moderate reduced activity 10 min - 4 h p.a. Otherwise, no abnormal clinical signs were observed.
A very slight till moderate erythema was observed in almost all of the animals for the entire observation period. Some animals had eschar on the skin and wrinkled skin. No signs of oedema were observed.
Body weight:
Weight gains were nonnal in all animals.
Gross pathology:
Gross pathological examinations at 14 days p. a. (terminal necropsies) revealed no test substance-dependent findings. Those macroscopic changes observed were attributable to the sacrificing procedure or to minor variations which often occur spontaneously in rats of this strain and age.
Interpretation of results:
GHS criteria not met
Conclusions:
Under the study conditions, the 14 d dermal LD50 was established at >2000 mg/kg bw in male and female rats.
Executive summary:

A study was conducted to determine dermal toxicity of the test substance according to OECD Guideline 402 and EC Directive 84/449. The acute dermal toxicity of the test substance was investigated in 5 male and 5 female Wistar rats. On the basis of the range finding results, each animal was given a single dermal administration of 2000 mg/kg bw as a 66.6% dilution. The pH was adjusted to 6.8 with a 25% ammonic solution. The skin was exposed to the test substance for 24 h and signs of erythema and oedema were subsequently evaluated once daily for 14 d. Clinical observations were conducted at regular intervals during the 14 d observation period. Body weights were measured on Days 0, 7 and 14. Gross pathological examinations were performed on animals at termination. No abnormal clinical signs were observed apart from a slight to moderate reduced activity in the animals. A very slight till well-defined erythema was observed in almost all of the animals for the entire observation period. Some animals had eschar on the skin and wrinkled skin 5 - 14 d post-administration (p.a.). No signs of oedema were observed. No pre-terminal deaths occurred. All animals showed normal weight gains. Gross pathological examinations at 14 d p.a. (terminal necropsy) revealed no test substance-dependent findings. Under the study conditions, the 14 d dermal LD50 was established at >2000 mg/kg bw in male and female rats (Kaufmann, 1990).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

Acute toxicity oral

A study was conducted to determine the potential toxic effect of the test substance when administered as a single oral dose to Wistar rats according to OECD Guideline 401. The test substance was administered once only, as supplied and at the dose levels of 360, 456, 564, 708 and 900 mg/kg bw, by the oral route (gastric gavage), in the Sprague-Dawley rat (5 males and 5 females per group). This study was performed in comparison with a control group of 10 rats of both sexes treated with the distilled water under the same conditions. Mortality and abnormal clinical signs were noted 15 min after intubation, then at 1, 2 and 4 h, and then daily for the 14 d study period. All the animals were weighed the day before treatment (Day-1), immediately before administration of the test substance (Day-1), on Days 8 and 15, as well as at time of death from Day-2 onwards. A necropsy was performed for all the animals dead during the study and for all surviving rats after the 14 d study period and the final observation (Day-15). Under the study conditions, the LD 50 of the test substance in the rat (male and female) was determined to be 730 mg/kg bw (651 - 819 mg/kg) by Bliss method and 735 mg/kg bw (641 – 844) by the Litchfield & Wilcoxon’s method (Lheritier, 1990).

Acute toxicity dermal

A study was conducted to determine dermal toxicity of the test substance according to OECD Guideline 402 and EC Directive 84/449. The acute dermal toxicity of the test substance was investigated in 5 male and 5 female Wistar rats. On the basis of the range finding results, each animal was given a single dermal administration of 2000 mg/kg bw as a 66.6% dilution. The pH was adjusted to 6.8 with a 25% ammonic solution. The skin was exposed to the test substance for 24 h and signs of erythema and oedema were subsequently evaluated once daily for 14 d. Clinical observations were conducted at regular intervals during the 14 d observation period. Body weights were measured on Days 0, 7 and 14. Gross pathological examinations were performed on animals at termination. No abnormal clinical signs were observed apart from a slight to moderate reduced activity in the animals. A very slight till well-defined erythema was observed in almost all of the animals for the entire observation period. Some animals had eschar on the skin and wrinkled skin 5 - 14 d post-administration (p.a.). No signs of oedema were observed. No pre-terminal deaths occurred. All animals showed normal weight gains. Gross pathological examinations at 14 d p.a. (terminal necropsy) revealed no test substance-dependent findings. Under the study conditions, the14 d dermal LD50 was established at >2000 mg/kg bw in male and female rats (Kaufmann, 1990).

Justification for classification or non-classification

The acute oral LD50 value of the test substance was determined to be between 300 and 2000 mg/kg bw in Sprague-Dawley rats. According to the EU CLP (EC 1272/2008) criteria, the substance therefore warrants a classification as Acute Tox. 4 – H302 (Harmful if swallowed).