1,1'-[methylenebis(oxy)]dibutane

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Butylal

IUPAC Name:

Butylal

Details on test material:

- Name of test material (as cited in study report): Butylal
- Physical state: colorless liquid
- Analytical purity: >99.99%
- Impurities (identity and concentrations): Formaldehyde (0.0038% w/w), water (not detected), n butanol (not detected).
- Composition of test material, percentage of components: Formaldehyde (0.0038% w/w), water (not detected), n butanol (not detected), butylal (>99.99% w/w)
- Lot/batch No.: E971205
- Expiration date of the lot/batch: No expiry date
- Container: one smoked glass flask
- Storage condition of test material: at room temperature and protected from light

The test substance was prepared in corn oil, batch No. 86H0059 (Sigma, 38297 Saint-Quentin-Fallavier, France).

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system 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

Administration / exposure

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).

Results and discussion

Effect levels

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:

other: 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

Gross pathology:

Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.

Other findings:

None

Applicant's summary and conclusion

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.