Registration Dossier

Administrative data

Description of key information

Oral

The calculated acute oral LD50 in female rats was 3362 mg/kg with 95 % confidence limits of 3002 to 3766 mg/kg.

Dermal

The LD50 value was found to be greater than 2000 mg/kg of body weight in male and female rabbits.

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:
27 February 1985 to 27 March 1985
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
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 and environmental conditions:
TEST ANIMALS
- Weight at study initiation: 180 - 280g
- Fasting period before study: 18 hours
- Housing: Rats were housed individually in stainless steel 0.5” wire mesh cages
- Diet: ad libitum, checked daily and added or replaced as needed. Feeders were designed to reduce soiling, bridging and scattering
- Water: ad libitum fresh tap water, fit for human consumption, using an automatic watering system
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30 to 70 %
- Photoperiod (hrs dark / hrs light): 12 hours of light, 12 hours of darkness
Route of administration:
oral: gavage
Vehicle:
other: 0.25 % Methylcellulose
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg (with the exception of the top dose level which was dosed as received at 4 mL/kg)
Doses:
1600, 2000, 2500, 3200, 3600 and 4000 mg/kg
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The rats were observed immediately and at one and four hours after dosing and twice daily for fourteen days for pharmacotoxicity, CNS effects and mortality. Body weights were recorded on the 14th day.
- Necropsy of survivors performed: Yes. The surviving rats were sacrificed by CO2 inhalation and a gross necropsy performed.
Statistics:
By the method of Litchfield and Wilcoxon
Sex:
male/female
Dose descriptor:
LD50
Effect level:
3 561 mg/kg bw
Based on:
test mat.
95% CL:
3 246 - 3 906
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
3 362 mg/kg bw
Based on:
test mat.
95% CL:
3 002 - 3 766
Mortality:
None of the rats died at 1600, 2000 or 2500 mg/kg; two of ten died at 3200 mg/kg, four of ten died at 3600 mg/kg and ten of ten died at 4000 mg/kg.
Clinical signs:
Signs observed included increased activity, decreased activity, nasal discharge, diarrhoea, salivation, lacrimation, ptosis, chromodacryorrhea, dyspnoea, decreased muscle tone, abnormal gait, abnormal stance and prostration.
Gross pathology:
Necropsy of animals dying on study revealed distended stomachs, lesions in the stomach mucosa and fluid-filled intestines. Congested, bright red lungs and foci on the kidneys were also observed.
No visible lesions were observed in any of the remaining animals at terminal necropsy.
Interpretation of results:
other: Not classified in accordance with EU criteria
Conclusions:
Under the conditions of the study, the LD50 in female rats was determined to be 3362 mg/kg with 95 % confidence limits of 3002 to 3766 mg/kg.
Executive summary:

The potential of the test material to cause acute toxicity via the oral route was investigated using methodology equivalent or similar to the standardised guideline OECD 401 under GLP conditions.

Following a dose range-finding study, six groups of ten Sprague Dawley rats (five males and five females) were administered the test material at dose levels of 1600, 2000, 2500, 3200, 3600 and 4000 mg/kg in 0.25 % methylcellulose. The rats were observed immediately and at one and four hours after dosing and twice daily for fourteen days for pharmacotoxicity, CNS effects and mortality. Body weights were recorded on the 14th day. The surviving rats were sacrificed by CO2 inhalation and a gross necropsy performed. The statistical method used was the method of Litchfield and Wilcoxon.

Signs observed included episodes of increased or decreased activity, nasal discharge, diarrhoea, salivation, lacrimation, ptosis, chromodacryorrhea, dyspnoea, decreased muscle tone, abnormal gait, abnormal stance and prostration. None of the rats died at 1600, 2000 or 2500 mg/kg; two of ten died at 3200 mg/kg, four of ten died at 3600 mg/kg and ten of ten died at 4000 mg/kg. Necropsy of the animals dying on study revealed distended stomachs, lesions in the stomach mucosa and fluid-filled intestines. Congested, bright red lungs and foci on the kidneys were also observed. No visible lesions were observed in any of the remaining animals at terminal necropsy.

The calculated acute oral LD50 in male and female rats was determined to be 3561 mg/kg with 95 % confidence limits of 3246 to 3906 mg/kg and a slope of 1.11. The calculated acute oral LD50 in females was determined to be 3362 mg/kg with 95% confidence limits of 3002 to 3766 mg/kg and a slope of 1.1. An attempt was made to calculate the LD50 in males but the data generated did not lend itself to the method of Litchfield and Wilcoxon.

Under the conditions of the study, the LD50 in female rats was determined to be 3362 mg/kg with 95 % confidence limits of 3002 to 3766 mg/kg.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
3 362 mg/kg bw
Quality of whole database:
The key study was conducted under GLP conditions utilising a method which is similar or equivalent to the standardised guideline OECD 401. It was assigned a reliability score of 1 in accordance with the criteria detailed by Klimisch et al. (1997) and the quality of the database is therefore considered to be good.

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
Study period:
05 November 1992 to 19 November 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Weight at study initiation: pre-test weight range was 2.2 to 2.4 kg for males and 2.1 to 2.3 kg for females
- Housing: One per cage in suspended wire mesh cages. Bedding was placed beneath the housing and changed twice per week.
- Diet: Fresh diet was provided daily
- Water: ad libitum
- Acclimation period: At least one week in quarantine

ENVIRONMENTAL CONDITIONS
- Temperature: Temperature controlled; value not reported
- Photoperiod: 12 hour dark/light cycle

IN-LIFE DATES
From: 20 October 1992
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: Dorsal area of the trunk (intact skin)
- % coverage: Approximately 10 % of the body surface
- Type of wrap if used: The test site was covered with a gauze patch, secured with non-irritating tape and gentle pressure was applied to the gauze to aid the distribution of the test material over the area. The torso was wrapped with plastic which was secured with non-irritating tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The test material was gently washed off with distilled water prior to dermal observations.
- Time after start of exposure: At 24 hours the patches were removed

TEST MATERIAL
- Amount(s) applied: The dose was based on the sample weight as calculated from the specific gravity
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed for toxicity and pharmacological effects at 1, 2 and 4 hours post dosing and once daily up to 14 days. Body weights were recorded pre-test, weekly and at death or termination. The animals were observed twice daily for mortality. The test sites were scored for dermal irritation at 24 hours post dose and on days 7 and 14 using the numerical Draize scale.
- Necropsy of survivors performed: Yes. All animals were examined for gross pathology. Abnormal tissues were preserved in 10 % buffered formalin for possible future examination.
Statistics:
The LD50, 95 % confidence limits, dose response curve and slope were calculated, if possible, by the method of Litchfield and Wilcoxon.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Nine out of ten animals survived the 2000 mg/kg dermal application. One male rabbit died on day 12.
Clinical signs:
The animal that died on day 12 exhibited pre-death physical signs of diarrhoea, lethargy and few faeces.
The nine out of ten animals that survived the 2000 mg/kg dermal application experienced instances of diarrhoea. Dermal reactions, absent to well defined on day 1 were absent on days 7 and 14.
Body weight:
Body weight changes were normal in all survivors.
Gross pathology:
Necropsy of the dead animal revealed abnormalities of the treated skin, lungs, pleural cavity and gastrointestinal tract. The death did not appear to be related to the treatment with the test material but to a pulmonary infection.
Necropsy results were normal in eight out of the nine surviving rabbits. One female exhibited liver abnormalities (nodules on liver).

Table 1: Body weight, dose volume and dermal reactions

Ear tag # & Sex

Dose volume (cc)

Body Weight (kg) on Day

Skin Reactions on Day

% Rem.

1

7

14

1

7

14

E

O

E

O

E

O

D4973-M

3.9

2.3

2.4

2.6

1a

2

0*

0

0*

0

70

D4969-M

4.1

2.4

2.1†

-

2a

2

0*

0

-

-

60

D4967-M

4.1

2.4

2.5

2.7

1a

1

0

0

0

0

40

D4971-M

3.9

2.3

2.3

2.4-

1a

1

0*

0

0*

0

50

D4963-M

3.7

2.2

2.3

2.3

1a

1

0

0

0

0

50

Mean

2.3

2.3

2.5

 

SD

0.08

0.15

0.18

Number

5

5

4

D5020-F

3.5

2.1

2.3

2.4

0a

0

0

0

0

0

40

D5011-F

3.5

2.1

2.3

2.4

1a

0

0f*

0

0

0

35

D5013-F

3.9

2.3

2.4

2.5

0a

0

0

0

0

0

40

D5017-F

3.9

2.3

2.4

2.6

0a

0

0*

0

0*

0

40

D5014-F

3.5

2.1

2.2

2.3

0a

0

0

0

0

0

35

Mean

2.2

2.3

2.4

 

SD

0.11

0.08

0.11

Number

5

5

5

†Body weight verified. This animal died on day 12 with a terminal body weight of 1.9 kg

E = erythema (redness)

O= oedema

a = dose site stained yellow

f = flaking skin

* = animal re-clipped

% Rem. = a visual estimate of the amount of material remaining on the skin, gauze and occlusive binding at 24 hours, after the occlusive binding was removed.

Interpretation of results:
other: Not classified in accordance with EU criteria
Conclusions:
Under the conditions of this study, the LD50 value was found to be greater than 2000 mg/kg of body weight.
Executive summary:

The potential of the test material to cause acute dermal toxicity in the rabbit was investigated in accordance with the standardised guideline OECD 402 under GLP conditions.

Five healthy male and five healthy female New Zealand Albino rabbits were dosed dermally with the test material at 2000 mg/kg of body weight. The test material was applied to intact skin and covered in an occlusive fashion for 24 hours. at the end of the exposure period, the test material was gently washed off with distilled water.

Animals were observed for toxicity and pharmacological effects at 1, 2 and 4 hours post dosing and once daily up to 14 days. Body weights were recorded pre-test, weekly and at death or termination. The animals were observed twice daily for mortality. The test sites were scored for dermal irritation at 24 hours post dose and on days 7 and 14 using the numerical Draize scale. All animals were examined for gross pathology.

Nine out of ten animals survived the 2000 mg/kg dermal application. One male rabbit died on day 12 and exhibited pre-death physical signs of diarrhoea, lethargy and few faeces.

The nine out of ten animals that survived the 2000 mg/kg dermal application experienced instances of diarrhoea. Dermal reactions, absent to well defined on day 1 were absent on days 7 and 14. Body weight changes were normal in all survivors.

Necropsy of the dead animal revealed abnormalities of the treated skin, lungs, pleural cavity and gastrointestinal tract. The death did not appear to be related to the treatment with the test material but to a pulmonary infection. Necropsy results were normal in eight out of the nine surviving rabbits. One female exhibited liver abnormalities (nodules on liver).

Under the conditions of this study, the LD50 value was found to be greater than 2000 mg/kg of body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
The key study was conducted under GLP conditions in accordance with a standardised guideline. It was assigned a reliability score of 2 in accordance with the criteria detailed by Klimisch (1997). The quality of the database is therefore considered to be good.

Additional information

Acute Oral Toxicity

The potential of the test material to cause acute toxicity via the oral route was investigated using methodology equivalent or similar to the standardised guideline OECD 401 under GLP conditions.

Following a dose range-finding study, six groups of ten Sprague Dawley rats (five males and five females) were administered the test material at dose levels of 1600, 2000, 2500, 3200, 3600 and 4000 mg/kg in 0.25 % methylcellulose. The rats were observed immediately and at one and four hours after dosing and twice daily for fourteen days for pharmacotoxicity, CNS effects and mortality. Body weights were recorded on the 14th day. The surviving rats were sacrificed by CO2 inhalation and a gross necropsy performed. The statistical method used was the method of Litchfield and Wilcoxon.

Signs observed included episodes of increased or decreased activity, nasal discharge, diarrhoea, salivation, lacrimation, ptosis, chromodacryorrhea, dyspnoea, decreased muscle tone, abnormal gait, abnormal stance and prostration. None of the rats died at 1600, 2000 or 2500 mg/kg; two of ten died at 3200 mg/kg, four of ten died at 3600 mg/kg and ten of ten died at 4000 mg/kg. Necropsy of the animals dying on study revealed distended stomachs, lesions in the stomach mucosa and fluid-filled intestines. Congested, bright red lungs and foci on the kidneys were also observed. No visible lesions were observed in any of the remaining animals at terminal necropsy.

The calculated acute oral LD50 in male and female rats was determined to be 3561 mg/kg with 95 % confidence limits of 3246 to 3906 mg/kg and a slope of 1.11. The calculated acute oral LD50 in females was determined to be 3362 mg/kg with 95% confidence limits of 3002 to 3766 mg/kg and a slope of 1.1. An attempt was made to calculate the LD50 in males but the data generated did not lend itself to the method of Litchfield and Wilcoxon.

Under the conditions of the study, the LD50 in female rats was determined to be 3362 mg/kg with 95 % confidence limits of 3002 to 3766 mg/kg.

Inhalation

In accordance with the Column 2 adaptation in Section 8.5 of Annex VIII of Regulation (EC) No 1907/2006 (REACH), it is considered justified to omit the acute toxicity testing by the inhalation route on the grounds that oral and dermal exposure are more likely.

Acute dermal toxicity

The potential of the test material to cause acute dermal toxicity in the rabbit was investigated in accordance with the standardised guideline OECD 402 under GLP conditions.

Five healthy male and five healthy female New Zealand Albino rabbits were dosed dermally with the test material at 2000 mg/kg of body weight. The test material was applied to intact skin and covered in an occlusive fashion for 24 hours. at the end of the exposure period, the test material was gently washed off with distilled water.

Animals were observed for toxicity and pharmacological effects at 1, 2 and 4 hours post dosing and once daily up to 14 days. Body weights were recorded pre-test, weekly and at death or termination. The animals were observed twice daily for mortality. The test sites were scored for dermal irritation at 24 hours post dose and on days 7 and 14 using the numerical Draize scale. All animals were examined for gross pathology.

Nine out of ten animals survived the 2000 mg/kg dermal application. One male rabbit died on day 12 and exhibited pre-death physical signs of diarrhoea, lethargy and few faeces.

The nine out of ten animals that survived the 2000 mg/kg dermal application experienced instances of diarrhoea. Dermal reactions, absent to well defined on day 1 were absent on days 7 and 14. Body weight changes were normal in all survivors.

Necropsy of the dead animal revealed abnormalities of the treated skin, lungs, pleural cavity and gastrointestinal tract. The death did not appear to be related to the treatment with the test material but to a pulmonary infection. Necropsy results were normal in eight out of the nine surviving rabbits. One female exhibited liver abnormalities (nodules on liver).

Under the conditions of this study, the LD50 value was found to be greater than 2000 mg/kg of body weight.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance does not require classification with respect to acute toxicity.