Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
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:
2001
Report Date:
2001

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Butylal
- Physical state: Colourless 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
- Storage condition of test material: at room temperature and protected from light
- Container: one smoked glass flask

Test animals

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

Administration / exposure

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

Results and discussion

Effect levels
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

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 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.