Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 09 Mar. 2006 to 23 Mar. 2006.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Test inspired from OECD guideline 402, with only one dose tested at 500 mg/Kg bw.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report Date:
2006

Materials and methods

Test guidelineopen allclose all
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratoires France, Domaine des Oncins, 69592 L'Arbresle Cedex, France.
- Age at study initiation: 8-12 weeks at the time of administration
- Weight at study initiation: In each group, individual weights of the animals did not deviate from the group mean weight by more than ± 20%. 217.1g, 228.9g and 220.9g
- Fasting period before study: period unknown
- Housing:Observations were performed at the time of delivery of the animals and daily during the period of acclimatisation. Animals were housed in cages of standard dimensions with sawdust bedding (SAFE, Reference B8/20). Cages were cleaned according CERB internal SOPs. The animals were placed in an air-conditioned (20-24 ° C) animal house kept at relative humidity between 45% and 65% (except during the cleaning slot) in which non-recycled filtered air was changed approximately 10 times per hour. The artificial day/night cycle involved 12 hours light and 12 hours darkness with light on at 7.30 a.m.
- Diet (e.g. ad libitum): RM1 (E)-SQC SDS/DIETEX feed (quality controlled/radiation sterilised) was available ad libitum except during the fasting experimental period. The criteria for acceptable levels of contaminants in the feed supplied were within the limits of the analytical specifications established by the diet manufacturer.
- Water (e.g. ad libitum): Drinking water was available ad libitum in polycarbonate feeder bottles with a stainless steel nipple. A specimen of water is obtained every 6 months and sent to the Laboratoire Departemental d'Analyse du Cher - 216, Rue Louis Mallet - 18014 Bourges Cedex, France, for analysis. The criteria for acceptable levels of contaminants in the water supplied were within the limits of the analytical specifications.
- Acclimation period: Minimum of five days before the treatment in the laboratory animal house where the experiment took place.

ENVIRONMENTAL CONDITIONS, see above

IN-LIFE DATES: From D1 day of administration to D15, day of euthanasia.

Administration / exposure

Type of coverage:
occlusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: the test item was applied to each animal on a piece of absorbant gauze measuring 30 cm2 (6 cm x 5 cm)
- % coverage: at least 10% of the total body surface
- Type of wrap if used: gauze was protected by a pad and covered with an adhesive tape Nylexfixr, for 24 hours

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period, any remaining test item was removed by rinsing with sterile water and the application site was gently rinsed with water.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 500 mg/kg body weight
- Concentration (if solution): Solid test item was moistened with sterile water 1mL/kg
- Constant volume or concentration used: yes
- For solids, paste formed: no

VEHICLE
- Amount(s) applied (volume or weight with unit): 1mL/kg
- Concentration (if solution):/
- Lot/batch no. (if required):/
- Purity:/
Duration of exposure:
24 hours
Doses:
At the Sponsor's request, a limit test at one dose level of 500 mg/kg body weight was carried out.
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality was recorded twice a day, i.e. in the morning and at the end of the working day. Animals were examined clinically on the day of treatment, 30 minutes, 2 hours, 4 hours and 6 hours after administration. Thereafter they were examined clinically at least once daily for 14 days. The skin examination was performed daily. Animals were weighed on D1, D7, D14 and D15
- Necropsy of survivors performed: yes. All animals surviving to the end of the 14-day monitoring period were euthanased on D15. All animals were subjected to gross necropsy and their organs (liver, spleen, kidneys, stomach, intestines, gonads/reproductive tract, lungs, heart, treated area and any other organs with obvious abnormalities) were examined macroscopically.
Statistics:
No statistical tests

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD0
Effect level:
500 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred during the study.
Clinical signs:
No clinical signs were observed during the course of the study.
Body weight:
Mean body weight and body weight gains values of treated animals on D7 were slightly lower than normal. Indeed, 2/3 females did not gain weight during this first week. Mean body weight and body weight gains values of treated animals on D14 were normal.
Gross pathology:
No organ or tissue gross findings were seen at necropsy. A redness was well perceptible on the application site of any animal only after exsanguination. It can be due to an irritation and/or a colouration induced by the test item.
Other findings:
On D2, after removal of the dressings, a purple colouring of the application site was observed in any animal which did not persist up to the end of the study. On D6 and D7, one animal presented a very slight erythema (barely perceptible; grade 1). No other dermal reactions were observed during the course of the study.

Any other information on results incl. tables

Effect on body weight of female rats (individual values)

Treatment  Animal number  D1  D7  D14  D15
500 mg/kg  200602869  217.1  217.6  235.5  217.8
 200602870  228.9  228.7  247.1  225.0
 200602871  220.9  231.3  248.2  229.1

Applicant's summary and conclusion

Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: other: Based on observed results
Conclusions:
Under the experimental conditions adopted in this study equivalent to an OECD 402, the maximal non-lethal dose of the test item was 500 mg/kg by dermal route in rats. Therefore test item was not considered as toxic by dermal route according to CLP criteria.
Executive summary:

At the Sponsor's request, the toxicity of the test item was evaluated after single dose administration by the dermal route in the rat inspired by the General Requirements of OECD Guideline No. 402 (February 24, 1987) and method B3 of Commission Directive No. 92/69/EEC (July 31, 1992) adapting to technical progress for the 17th time Council Directive No. 67/548/EEC and subsequent amendments (GLP study, scored as validity 2 according to Klimisch criteria). At the Sponsor's request, a single group of 3 females was dosed at 500 mg/kg body weight. The test item was applied on the skin in the dorsal region of each animal on an area exposing at least 10 % of the total body surface. The test item was moistened with sterile water. The absorbent gauze was held in place for 24 hours using an elastic band.

Under the experimental conditions adopted, the dermal application of the test item at 500 mg/kg body weight caused no mortality and a very slight erythema on D6 and D7 was observed in 1/3 female Sprague-Dawley rats. Under the experimental conditions adopted, the maximal non-lethal dose of the test item was 500 mg/kg by dermal route in rats.