Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: An apparently well done GLP study performed according to OECD test guideline 402.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1994
Report Date:
1994

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
not specified
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
gas under pressure: refrigerated liquefied gas
Details on test material:
- Name of test material (as cited in study report): PEROXIMON K3
- Molecular formula (if other than submission substance): C5H10O4
- Molecular weight (if other than submission substance): 134.6
- Physical state: Clear Liquid
- Analytical purity: 35% max (W/W)
- Impurities (identity and concentrations):
- Composition of test material, percentage of components: Acetylacetate peroxide, 35%
- Purity test date: July, 1, 1994.
- Lot/batch No.: Protocol: RN 0331; Labelling: RN 0331; (30/3/1994); Elf Atochem filing Number : CAL 2658/94
- Expiration date of the lot/batch: June 1995
- Specific gravity: 1.025
- Storage condition of test material: In dark and 4 degrees Celcius
- Other: Soluble > 50 g/100 ml in water; > 50 g/ml in methylic alcohol

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Animals
Species, strain: rat, Sprague-Dawley ICO: OFA-SD (lOPS Caw).
Reason for this choice: rodent species cOIl11ilonly requested by the international regulations for this type of study.
Breeder: Iffa Credo, 69210 L' Arbresle, France.
Number and sex: 1 group of 10 animals (5 males and 5 females).
Age/weight: on the day of treatment, the animals were approximately 8 weeks old, and had a mean body weight of 256 ± 12 g for the males and 219 ± 4 g for the females.
Acclimatization: at least 5 days before the beginning of the study.
Identification of the animals: the animals were identified individually by earmarks or earnotches.

Environmental conditions
During the acclimatization period and during the main test, the conditions in the animal room were as follows:
temperature: 21 ± 2°C
relative humidity: 30 to 70%
light/dark cycle: 12 hl12 h
ventilation: about 12 cycleslhour of filtered, non-recycled air.
The temperature and relative humidity were recorded continuously and records retained.
The housing conditions (temperature, relative humidity, light/dark cycle and ventilation) were checked monthly. The animals were housed in polycarbonate cages covered with a stainless steel lid.
During the acclimatization period each cage (48 x 27 x 20 cm) contained 4 to 7 animals of the same sex. During the treatment period each cage (35.5 x 23.5 x 19.3 cm) contained 1 rat. Each cage
contained graded, dust-free sawdust (SICSA, 94142 Alfortville, France). Bacteriological analysis of the sawdust and detection of possible contaminants (pesticides, heavy metals) are performed periodically.

Food and water
All the animals had free access to A04 C pelleted diet (U.A.R., 91360 Villemoisson-sur-Orge, France). Each batch of food was analysed (composition and contaminants) by the supplier. The diet formula is presented in appendix 2.

Drinking water filtered by a F.G. Millipore membrane (0.22 micron) was contained in bottles. Bacteriological and chemical analysis of the water and detection of possible contaminants (pesticides, heavy metals and nitrosamines) are performed periodically. Results are archived at C.LT. There were no contaminants in the diet, water or sawdust at levels likely to have influenced the
outcome of the study.

Administration / exposure

Type of coverage:
occlusive
Details on dermal exposure:
TREATMENT

Preparation of the animals
On the day before treatment, the dorsal area (6 cm x 8 cm) of each animal was clipped of hair using electric clippers. Only animals with healthy intact skin were used for the study.

Mode of treatment
As the test substance was anticipated to be non-toxic at 2000 mg/kg, a limit test was performed by application of 2000 mg/kg of the test substance to a group of 10 animals (5 males and 5 females).

Taking into consideration that the specific gravity (SO) of the test substance was 1.025, the test substance in its original form and at a dose of 2000 mg/kg, was applied directly to an area of the skin representing approximately 10% (5 x 6 cm for the females and 5 x 7 cm for the males) of the body surface of the animals. This was calculated according to Meeh's formula (1). A hydrophilic gauze pad (Semes France, 54183 Heillecourt, France) was then applied to the skin. The test substance and the gauze pad were held in contact with the skin for 24 hours by means of an adhesive hypoallergenic aerated semi-occlusive dressing (Laboratoires de Pansements et d'Hygiene, 21300 Chenove, France) and a restraining bandage (Laboratoires 3M Sante, 92245 Malakoff, France). This dressing prevented ingestion of the test substance by the animals. No residual test substance was observed at removal of the dressing.
Duration of exposure:
Date of treatment
The single administration was performed on 1.9.94 in the morning (day 1) and was followed by
a 14-day observation period until 15.9.94 (day 15).
Doses:
As the test substance was anticipated to be non-toxic at 2000 mg/kg, a limit test was performed by application of 2000 mg/kg of the test substance to a group of 10 animals (5 males and 5 females). The dose applied to each animal was adjusted according to body weight determined on the day
of treatment.
No. of animals per sex per dose:
5
Control animals:
yes
Details on study design:
CLINICAL EXAMINATIONS

Clinical signs
The animals were observed frequently during the hours following application of the test substance for detection of possible treatment-related clinical signs. Observation of the animals was made at least once a day for a period of 14 days, to determine whether any of the clinical signs were reversible or not.

Mortality
The animals were checked frequently during the hours following application of the test substance for mortality or signs of morbidity, then at least twice a day thereafter.

(1) Meeh's formula (AFNOR-T03-23 Standard, August 1980, France)
S =k\j"fT
S = Total surface of the animal in dm2
p = Weight of the animal in grams
k =0.09 for the rat


Body weight
The animals were weighed individually just before administration of the test substance then on days 8 and 15. The body weight gain of the treated animals was compared to a reference curve of C.LT. control animals with the same initial weight.

PATHOLOGY

Necropsy
On day 15, the animals were killed by CO2 inhalation in excess and a macroscopic examination was performed.

Macroscopic examination
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.

Microscopic examination
No microscopic examination was performed.
Statistics:

DATA EVALUATION

Evaluation of the innoxiousness or toxicity of the test substance following a single dermal application in rats should include the relationship, if any, between the animals' exposure to the test substance and the incidence and severity of all the abnormalities including behavioural and clinical abnormalities, macroscopic lesions, body weight changes, mortality and any other toxic effects.

Classification of the test substance is based on the following criteria: Commission Directive 93/21/E.E.C.


LDso
dermal route Labelling Indication Symbol
(mg/kg) sentence of danger


<50 R27 Very toxic in contact with skin Very toxic T+
50 < and < 400 R24 Toxic in contact with skin Toxic T
400 < and < 2000 R21 Harmful in contact with skin Harmful Xn
>2000 None None None

Results and discussion

Preliminary study:
Results
There was no preliminary study.

Results of the main study:
The general behaviour and body weight gain of the animals were not affected by treatment with the test substance. No cutaneous reactions were observed. No deaths occurred at 2000 mg/kg. A macroscopic examination revealed no abnormalities in the animals killed at the end of the study.
Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Mortality:

Mortality:
No deaths occurred during the observation period.
Clinical signs:
CLINICAL EXAMINATIONS
Clinical signs and cutaneous reactions (tables 1 and 2): No clinical signs and cutaneous reactions were observed during the study.
Body weight:
Body weight (treated animals and control animals)

Between days 1 and 8, a decrease in body weight was noted for one female (No. 01). This finding was probably due to the stress caused by the experimental conditions and was not attributed to treatment with the test substance. The body weight of this animal showed an improvement during the remainder of the observation period. The body weight gain of the remaining animals was normal.
Gross pathology:
PATHOLOGY

Macroscopic examination of the main organs of the animals killed at the end of the study revealed no apparent abnormalities.

Applicant's summary and conclusion

Conclusions:
CONCLUSION

Under the experimental conditions, the LDo of the test substance, PEROXIMON K3, when administered by dermal route in rats was higher than or equal to 2000 mglkg. No signs of toxicity were observed at this dose.
Executive summary:

SUMMARY

At the request of Elf Atochem, Milano, Italia, the acute TOXICITY of the test substance, PEROXIMON K3, by dermal route was evaluated in rats according to O.E.C.D. (No. 402, 24th February 1987) and E.C. (92/69/E.E.C.). The study was conducted in compliance with the

principles of Good Laboratory Practice.

 

Methods

The test substance was administered by dermal route to a group of 10 Sprague-Dawley rats (5 males and 5 females). The test substance in its original form was applied directly to the skin of the animals at a dose of 2000 mg/kg, taking into consideration that the specific gravity (SG) of the test substance was 1.025. After 24 hours under a semi-occlusive dressing, no residual test substance was observed on removal of the dressing. The animals were checked for clinical signs, mortality and body weight gain for a period of 14 days following the single application of the test substance. A necropsy was performed on each animal killed at the end of the study.

 

Results

The general behaviour and body weight gain of the animals were not affected by treatment with the test substance. No cutaneous reactions were observed. No deaths occurred at 2000 mg/kg. A macroscopic examination revealed no abnormalities in the animals killed at the end of the study.

 

Conclusion

Under the experimental conditions, the LD50 of the test substance, PEROXIMON K3, when administered by dermal route in rats was higher than or equal to 2000 mg/kg. No signs of toxicity were observed at this dose.