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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
Reliability:
1 (reliable without restriction)
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Breeder: Iffa Crédo, 69210 L'Arbresle, France.
- Age at study initiation: The animals were approximately 6 weeks old.
- Weight at study initiation: The animals had a mean body weight ± standard deviation of 181 ± 12 g for the males and 143 ± 6 g for the females.
- Fasting period before study: The animals were fasted for an overnight period of approximately 18 hours before dosing, but had free access to water.
Food was given back approximately 4 hours after administration of the test substance.
- Housing: The animals were housed in polycarbonate cages (48 cm x 27 cm x 20 cm). Each cage contained four to seven animals of the same sex during the acclimatization period and five rats of the same sex during the treatment period. Each cage contained dust-free sawdust (SICSA, 94142 Alfortville, France). Bacteriological and chemical analysis of the sawdust, including the detection of possible contaminants (pesticides, heavy metals), are performed regularly by external laboratories. The results of these analyses are archived at CIT.
- Diet (e.g. ad libitum): All the animals had free access to A04 C pelleted diet (UAR, 91360 Villemoisson-sur-Orge, France), except during fasting period. Each batch of food was analysed by the supplier for composition and contaminant levels.
- Water (e.g. ad libitum): Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum. Bacteriological and chemical analysis of the water and diet, including the detection of possible contaminants (pesticides, heavy metals and nitrosamines), are performed regularly by external
laboratories. No contaminants are known to be present in the diet, drinking water or bedding material at levels which may be expected to interfere with or prejudice the outcome of the study.
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2
- Humidity (%): 30 to 70
The temperature and relative humidity were under continuous control and recording. The records were checked daily and retained. In addition to these daily checks, the housing conditions and corresponding instrumentation and equipment are verified and calibrated at regular intervals.

- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air.
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: not specified
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: not specified
- Amount of vehicle (if gavage): The test substance was prepared in corn oil and administered to the animals under a volume of 10 ml/kg.
- Justification for choice of vehicle: not specified
- Lot/batch no. (if required): 86H0059 (Sigma, 38297 Saint-Quentin-Fallavier, France).
- Purity: not specified

Doses:
2000; 5000; 7000; 10000 mg/kg
No. of animals per sex per dose:
5 males for doses of 2000 and 10000 mg/kg
5 females for doses of 2000; 5000; 7000; 10000 mg/kg
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
The animals were observed frequently during the hours following administration of the test substance, for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day until day 15. Type, time of onset and duration of clinical signs were recorded for each animal individually. Time of death was recorded individually, in terms of the number of hours or days after dosing.

The animals were weighed individually just before administration of the test substance on day 1 and then on days 8 and 15.
Individual weights of animals found dead during the study were measured at necropsy when survival exceeded 24 hours and if no signs of "cannibalism" were present. The body weight gain of the treated animals was compared to that of CIT control animals with the same initial body weight.
- Necropsy of survivors performed: yes
The animals found dead during the study were subjected to a macroscopic examination as soon as possible.
On day 15, all surviving animals were killed by carbon dioxide asphyxiation and a macroscopic examination was performed.
After opening the thoracic and abdominal cavities, a macroscopic examination of the main organs (digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities) was performed.
In case of macroscopic lesions, organ samples were taken and preserved in 10% buffered formalin.
No microscopic examination was performed.
- Other examinations performed: none
Statistics:
The LD50 value, expressed in milligrams of test substance per kilogram of animal (mg/kg), was calculated according to Probit-Analysis (Weber (1972) and Bliss (1938)).
The 70 to 95% confidence interval limits were calculated statistically according to Fieller's method (1944).
Sex:
male/female
Dose descriptor:
LD50
Effect level:
6 873 mg/kg bw
Based on:
test mat.
95% CL:
2 184 - 19 981
Remarks on result:
other: with 95% confidence interval limits
Mortality:
Mortality was as follows (number of animals which died/total number of animals in the group):
At dose of 2000 mg/kg: 0/5 for male and 0/5 for female.
At dose of 5000 mg/kg: 0/5 for female (not tested on male).
At dose of 7000 mg/kg: 5/5 for male and 5/5 for female.
At dose of 10000 mg/kg: 3/5 for female (not tested on male).
Clinical signs:
At the 10000 mg/kg dose-level, 3/5 females died between days 2 and 4. Hypoactivity or sedation, coma, piloerection, dyspnoea, lateral recumbency and unsteady gait were observed from day 1 in almost all animals. Stiff gait, ptosis and ocular secretion were also noted between days 6 and 13 in 1/2 surviving animals. Recovery was complete in the surviving animals on day 4 or 14.
At the 7000 mg/kg dose-level, all animals died between days 1 and 5. Hypoactivity or sedation, coma, piloerection, dyspnoea, lateral recumbency and unsteady gait were observed prior to death in almost all animals.
At the 5000 mg/kg dose-level, no mortality occurred. Hypoactivity or sedation, piloerection, lateral recumbency, unsteady gait, stiff gait, tremors and coma were observed from day 1 in almost all animals. Recovery was complete in all animals on day 8.
At the 2000 mg/kg dose-level, no mortality occurred. On day 1, hypoactivity and piloerection were observed in all animals; unsteady gait was also noted in one of these animals. Recovery was complete in all animals on day 2.
Body weight:
The body weight gain of the animals given 5000 mg/kg and of the surviving animals given 10000 mg/kg was reduced during the first week of the study. The body weight gain of the animals given 2000 mg/kg was not affected by treatment with the test substance.
Gross pathology:
Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.
Other findings:
None
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under experimental conditions, the oral LD50 of the test substance BUTYLAL is, for female rats, 6873 (2184 - 19981) mg/kg with 95% confidence interval limits. Toxicity is comparable in males.
Executive summary:

The acute oral toxicity of the test substance BUTYLAL was evaluated in rats according to OECD (No. 401, 24th February 1987) and EC (92/69/EEC, B.1, 31st July 1992) guidelines. The study was conducted in compliance with the Principles of Good Laboratory Practice Regulations.

The test substance was administered by oral route (gavage) to groups of five male and/or five female fasted Sprague-Dawley rats. The test substance was prepared in corn oil and administered to the animals under a volume of 10 ml/kg. The study design was as follows. Doses of 2000; 5000; 7000; 10000 mg/kg was tested with 5 males for doses of 2000 and 10000 mg/kg 5 females for doses of 2000; 5000; 7000; 10000 mg/kg. Vehicle used was corn oil at 10 ml/kg.

Clinical signs, mortality and body weight gain were checked for a period of up to 14 days following the single administration of the test substance. All animals were subjected to necropsy. The LD50 was calculated for females according to Probit's method. The interpretation of results was carried out according to the classification criteria laid down in Council Directive 93/21/EEC (27th April 1993) adapting to technical progress for the eighteenth time Council Directive 67/548/EEC.

At the 10000 mg/kg dose-level, 3/5 females died between days 2 and 4. Hypoactivity or sedation, coma, piloerection, dyspnoea, lateral recumbency and unsteady gait were observed from day 1 in almost all animals. Stiff gait, ptosis and ocular secretion were also noted between days 6 and 13 in 1/2 surviving animals. Recovery was complete in the surviving animals on day 4 or 14. At the 7000 mg/kg dose-level, all animals died between days 1 and 5. Hypoactivity or sedation, coma, piloerection, dyspnoea, lateral recumbency and unsteady gait were observed prior to death in almost all animals.

At the 5000 mg/kg dose-level, no mortality occurred. Hypoactivity or sedation, piloerection, lateral recumbency, unsteady gait, stiff gait, tremors and coma were observed from day 1 in almost all animals. Recovery was complete in all animals on day 8. At the 2000 mg/kg dose-level, no mortality occurred. On day 1, hypoactivity and piloerection were observed in all animals; unsteady gait was also noted in one of these animals. Recovery was complete in all animals on day 2.

Mortality was as follows (number of animals which died/total number of animals in the group). At dose of 2000 mg/kg: 0/5 for male and 0/5 for female. At dose of 5000 mg/kg: 0/5 for female (not tested on male). At dose of 7000 mg/kg: 5/5 for male and 5/5 for female. At dose of 10000 mg/kg: 3/5 for female (not tested on male).

The body weight gain of the animals given 5000 mg/kg and of the surviving animals given 10000 mg/kg was reduced during the first week of the study. The body weight gain of the animals given 2000 mg/kg was not affected by treatment with the test substance. No apparent abnormalities were observed in all animals at necropsy.

Under our experimental conditions, the oral LD50 of the test substance BUTYLAL is, for female rats, 6873 (2184 - 19981) mg/kg with 95% confidence interval limits. Toxicity is comparable in males.

According to the classification criteria laid down in Commission Directive 93/21/EEC, concerning the potential toxicity by oral route, the test substance should not be classified.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
6 873 mg/kg bw
Quality of whole database:
Acute oral toxicity was studied in an OECD 401 and GLP test.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
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)
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
yes (incl. certificate)
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: Breeder: Iffa Crédo, 69210 L'Arbresle, France.
- Age at study initiation: The animals were approximately 8 weeks old, and
- Weight at study initiation: The animals had a mean body weight ± standard deviation of 261 ± 7 g for the males and 228 ± 14 g for the females.
- Fasting period before study: The animals were fasted for an overnight period of approximately 18 hours before dosing, but had free access to water.
Food was given back approximately 4 hours after administration of the test substance.
- Housing: During the acclimatization period, the animals were housed in polycarbonate cages (48 cm x 27 cm x 20 cm). During the treatment period, the animals were housed individually in polycarbonate cages (35.5 cm x 23.5cm x 19.3 cm). Each cage contained dust-free sawdust (SICSA, 94142 Alfortville, France). Bacteriological and chemical analysis of the sawdust, including the detection of possible contaminants (pesticides, heavy metals), are performed regularly by external laboratories. The results of these analyses are archived at CIT.
- Diet (e.g. ad libitum): All the animals had free access to A04 C pelleted diet (UAR, 91360 Villemoisson-sur-Orge, France), except during fasting period. Each batch of food was analysed by the supplier for composition and contaminant levels.
- Water (e.g. ad libitum): Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum. Bacteriological and chemical analysis of the water and diet, including the detection of possible contaminants (pesticides, heavy metals and nitrosamines), are performed regularly by external
laboratories. The results of these analyses are archived at CIT.
No contaminants are known to be present in the diet, drinking water or bedding material at levels which may be expected to interfere with or prejudice the outcome of the study.
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2
- Humidity (%): 30 to 70
The temperature and relative humidity were under continuous control and recording. The records were checked daily and retained. In addition to these daily checks, the housing conditions and corresponding instrumentation and equipment are verified and calibrated at regular intervals.
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air.
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: not speciifed
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsal area
- % coverage: 10% (5 cmx 6cm for females and 5 cm x 7 cm for males)
- Type of wrap if used: An hydrophilic gauze pad (Coopérative Pharmaceutique Française, 77000 Melun, France) was applied to the skin. The test substance and the gauze pad were held in contact with the skin for 24hoursby means of an adhesive hypoallegenic aerated semi-occlusive dressing (Laboratoires de Pansements et d'Hygiène, 21300 Chenove, France) and a restraining bandage (Laboratoires 3M Santé, 92245 Malakoff, France). This dressing prevented ingestion of the test substance by the animals.

REMOVAL OF TEST SUBSTANCE
No residual test substance was observd at removal of the dressing.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg butylal/kg bw. The dose applied to each animal was adjusted according to the body weight determined on the day of treatment.
- Concentration (if solution): 2000 mg/kg bw
- Constant volume or concentration used: yes/no


VEHICLE
Taking into consideration that the density of the test substance was 0.83, the test substance was applied in its original form without vehicle at the dose volume of 2000 mg/kg bw.
Duration of exposure:
14 days
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 males and 5 females
Control animals:
other: CIT historical control rats were used for body weight comparison with treated rats. CIT historical data of animals dosed by the dermal route was obtained from results of control animals from October 1995 to December 1997.
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
The animals were observed frequently during the hours following administration of the test substance, for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day until day 15.
Type, time of onset and duration of clinical signs were recorded for each animal individually.
Time of death was recorded individually, in terms of the number of hours or days after dosing.
The animals were weighed individually just before administration of the test substance on day 1 and then on days 8 and 15.
The body weight gain of the treated animals was compared to that of CIT control animals with the same initial body weight.

- Necropsy of survivors performed: yes
On day 15, all surviving animals were killed by carbon dioxide asphyxiation and a macroscopic examination was performed.
After opening the thoracic and abdominal cavities, a macroscopic examination of the main organs (digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities) was performed.
In case of macroscopic lesions, organ samples were taken and preserved in 10% buffered formalin.
No microscopic examination was performed.
- Other examinations performed: clinical signs, body weight, macroscopic examination of the main organs
Statistics:
Not necessary
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No death occured during the observation period.
Clinical signs:
No clinical signs and no cutaneous reactions were observed during the study.
Body weight:
The body weight gain of females was slightly reduced when compared tohistorical control animals. The body weight gain of females was not affected by treatment with the test substance.
Gross pathology:
Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.
Other findings:
None
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under experimental conditions, the dermal LD50 of the test substance BUTYLAL is higher than 2000 mg/kg bw in rats.
Executive summary:

The acute dermal toxicity of the test substance BUTYLAL was evaluated in rats according to OECD (No. 402, 24th February 1987) and EC (92/69/EEC, B.3, 31st July 1992) guidelines. The study was conducted in compliance with the Principles of Good Laboratory Practice Regulations.

The test substance was applied to the skin of one group of ten Sprague-Dawley rats (five males and five females). The application was performed with the undiluted test substance at the dose of 2000 mg/kg, taking into consideration that its density was 0.83. The test site was then covered by a semi-occlusive dressing for 24 hours. Clinical signs, mortality and body weight were checked for a period of 14 days following the single administration of the test substance.

All animals were subjected to necropsy. The interpretation of results was carried out according to the classification criteria laid down in Council Directive 93/21/EEC (27th April 1993) adapting to technical progress for the eighteenth time Council Directive 67/548/EEC.

No death occured at 2000 mg/kg. The general behaviour of the animals was not affected by treatment with the test substance. The body weight gain of females was slightly reduced when compared to historical control animals. The body weight gain of males was not affected by treatment with the test substance. No cutaneous reaction were observed. No apparent abnormalities were observed at necropsy.

Under our experimental conditions, the dermal LD50 of the test substance BUTYLAL is higher than 2000 mg/kg in rats. According to the classification criteria laid down in Commission Directive 93/21/EEC, concerning the potential toxicity by dermal route, the test substance should not be classified.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Acute dermal toxicity was studied in an OECD 402 and GLP test.

Additional information

Justification for selection of acute toxicity – oral endpoint

The oral LD50 of the test substance BUTYLAL is, for female rats, 6873 (2184 - 19981) mg/kg with 95% confidence interval limits.

Justification for selection of acute toxicity – inhalation endpoint

In accordance with column 2 of REACH Annex VIII relative to Acute toxicity (Point 8.5), in addition to the oral route (8.5.1), for substances other than gases, the information mentioned under 8.5.2 (acute toxicity by inhalation) to 8.5.3 (acute toxicity by derma route) shall be provided for at least one other route. The choice for the second route will depend on the nature of the substance and the likely route of human exposure. If there is only one route of exposure, information for only that route need be provided.

Testing by dermal route is appropriate regarding to the test substance exposure and physicochemical and toxicological properties.

Testing by inhalation is not appropriate.

Justification for selection of acute toxicity – dermal endpoint

The dermal LD50 of the test substance BUTYLAL is higher than 2000 mg/kg bw in rats

Justification for classification or non-classification

According to the classification criteria laid down in Commission Directive 93/21/EEC, concerning the potential toxicity by oral route, the test substance should not be classified.

According to the classification criteria laid down in Commission Directive 93/21/EEC, concerning the potential toxicity by dermal route, the test substance should not be classified.