Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
December 1 - December 18, 2009
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name: ISOSORBIDE C PHARMA
Supplier: ROQUETTE FRERES
Batch Number: E1143
CAS number: 652-67-5
Purity: 99.8%
Moisture: 0.3%
Appearance: solid flakes
Odour: neutral
Molecular formula: C6H10O4
Molecular weight: 146.16 g/mol
Solubility: highly soluble in water
pH: 5.0 - 7.0
Stability: stable with respect to storage conditions

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Sex: Male and female. Females were nulliparous and non-gravid.
Origin: Charles River Laboratoires France, Domaine des Oncins, 69592 L0Arbresle Cedex, France.
Identification: Animals were identified individually by marking the tail with a felt-tip marker.
Age: 8 weeks at the time of administration.
Weight: 175.7 g and 300.5 g at the start of the experiment.
Number: 5 males, 5 females.

Housing: Daily observations were undertaken at the time of delivery of the animals and during the period of acclimatisation. Animals were housed in cages of standard dimensions with sawdust bedding.
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. Any deviations outside of the temperature or hygrometry ranges
were reported by the Study Director according to the SOP 5.36. The artificial day/night cycle involved 12 hours light and 12 hours darkness with light on at 7.30 a.m..
Feeding: 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 supply were within the limits of the analytical specifications established by the diet manufacturer.
Drinking water: Drinking water was available ad libitum in polycarbonate bottles with a stainless steel nipple. A specimen of water is obtained every 6 months and sent to IPL (Institut Pasteur de Lille-Laboratoire d0analyses de l0Allier) - Boulevard Nomazy - BP1707 - 03017 Moulins Cedex - France, for analysis. The criteria for acceptable levels of contaminants in the water supplied were within the limits of the analytical specifications.

Administration / exposure

Type of coverage:
occlusive
Vehicle:
water
Remarks:
Sterile
Details on dermal exposure:
TEST SITE
ISOSORBIDE was applied uniformly to the skin in the dorsal region of each animal on a previously shaved area without prior fasting. For this purpose, on the day prior to application, the dorsal region of each rat was carefully clipped on an area of approximately 50 cm2 using an electric clipper, avoiding damage to the skin.
On the day of study, test item was applied to the prepared area to expose at least 10% of the total body surface. ISOSORBIDE was applied to each animal on a piece of absorbant gauze measuring 30 cm2 (6 cm x 5 cm) which was protected by a pad and covered with an adhesive tape Nylexfixr. The absorbent gauze was held in place for 24 hours except for Animal No. 200905841 which was found without pad.

Duration of exposure:
72 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 males, 5 females.
Control animals:
no
Details on study design:
Findings are routinely noted individually on clinical case report forms.
Mortality was recorded twice a day, i.e. in the morning and at the end of the working day.

General observations: Animals were examined clinically before treatment, twice on the day of treatment (60 minutes ± 30 minutes after administration and then again between 3 and 4 hours post-dose). Thereafter they were examined clinically at least once daily for 14 days.
Full clinical examination: On D2, at least 1 hour after removal of the dressings, and on D7 the animals were submitted to a full clinical examination outside the housing cage, including functional and neurobehavioural tests .
Skin lesion evaluation: The skin examination was performed daily.
Animals were weighed on the following days:
• on D1 (day of administration)
• on D7 and D14 during the study
• on D15, day of euthanasia.
Food and water consumption was not measured.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 other: mg/kg
Based on:
test mat.
Mortality:
No mortality occurred during the study
Clinical signs:
No clinical sign was observed during the course of the study.
Body weight:
Mean weight gain in treated animals was normal when compared with the hystorical range of values.
Gross pathology:
No organ or tissue gross findings were seen at necropsy
Other findings:
No dermal reaction was observed during the course of the study

Any other information on results incl. tables

The application of the test item did not induce colouration of the application site and did not interfere with the grading of any skin lesion. Under the experimental conditions adopted, no mortality occurred during the study. No clinical signs and no dermal reactions were observed during the course of the study. Mean weight gain in treated animals was normal when compared with the range of values usually found in our Centre. No organ or tissue gross findings were seen at necropsy.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The LD50 of the test item ISOSORBIDE administered by the dermal route was higher than 2000 mg/kg in the male and female Sprague-Dawley rat.
Based on the results of this study, Isosorbide was not classified according to Regulation No.1272/2008.