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: GLP study in compliance with guidelines

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- public name of test material:
Reaction mass of 6,13-dichloro-3,10-bis{[2-({[(2-chloroethyl)sulfonyl]alkanoyl}amino)ethyl]-amino}- polycarboheterocyclo 4,11-disulfonic acid, mono and/or disodium salt and 6,13-dichloro-3-{[2-({[(2-chloroethyl)sulfonyl]alkanoyl}amino) ethyl]amino}-10-[(2-{[4-(ethenylsulfonyl)alkanoyl]amino}ethyl)amino] polycarboheterocyclo -4,11-disulfonic acid, mono and/or di sodium salt
- Physical state: solid, dark blue powder
- Analytical purity: approx. 86.6%
- Lot/batch No.: VER 2108 BOP 02/07
- Expiration date of the lot/batch: November 30, 2012
- Storage condition of test material: At room temperature, light protected, kept strictly dry
- Other:

Test animals

Species:
rat
Strain:
other: HanRcc: WIST(SPF)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories Ltd, Laboratory Animal Services, Füllinsdorf, Switzerland
- Age at study initiation: Males were 9 weeks, females were 11 weeks
- Weight at study initiation: 187.2 to 260.1 g
- Fasting period before study:
- Housing: During acclimatisation in groups of five in Makrolon type-4 cages with standard softwood bedding; individually in Makrolon type-3 cages with softwood bedding (Lignocel, Schill AG, Muttenz, Switzerland)
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 44/04 (Provimi Kliba AG, Kaiseraugst, Switzerland) ad libitum
- Water (e.g. ad libitum): Community tab water from Füllinsdorf ad libitum
- Acclimation period: Under laboratory conditions after health examination

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 °C
- Humidity (%): 30 to 70%
- Air changes (per hr): 10 to 15
- Photoperiod (hrs dark / hrs light): 12 hours light, 12 hours darkness

IN-LIFE DATES: From: To:

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
other: purified water (deinoised, processed with PURELAB Option-R unit using reverse osmosis, adsorption, ion-exchange and photo oxidation)
Details on dermal exposure:
TEST SITE
- Area of exposure: backsof the animals
- % coverage: approximately 10% of total body surface area
- Type of wrap if used: elastic adhesive bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): flushed with lukewarm tap water
- Time after start of exposure: 24 hours

TEST MATERIAL and VEHICLE
Test material was weighed and purified water was added. Approximately 6 mL/kg bodyweight of the mixture was evenly applied to the back of testing animals using a syringe, so that the final dose was 2000 mg/kg bodyweight.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bodyweight
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Viability and mortality after 30 minutes, 1, 2, 3 and 5 hours at day of application and twice daily at study days 2 to 15
Clinical signs after 30 minutes, 1, 2, 3 and 5 hours at day of application and once daily at study days 2 to 15
Local dermal signs once daily on study days 2 and 15
Bodyweights on study days 1, 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:
Statistics:
No statistical analysis was used.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths occurred during the study
Clinical signs:
No clinical signs were observed in the course of the study
Body weight:
One female did not gain bodyweight during the first week after treatment but recovered until the end of the study. The bodyweight of the other animals was within the range commonly recorded for this strain and age.
Gross pathology:
No macroscopic findings were recorded at necropsy
Other findings:
A marked blue staining produced by the test item was observed in all males and females at removal of dressing and persisted as slight up to test days 10, 13 or 15. The erythema was not assessable due to the blue staining up to test days 4, 7 or 8. When assessable, no reddening was observed

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
No adverse effects were seen in male and female rats receiving a single application of test material dissolved in water at a dose of 2000 mg/kg bodyweight under semiocclusive conditions for 24 hours.
Executive summary:

The acute dermal toxicity of test material was tested in a GLP study in accordance with EU Method B.3. A single dose of 2000 mg/kg bodyweight of the test substance dissolved in water was applied topically to the backs of five male and five female adult HanRcc WIST(SPF) rats. The application site was kept under semiocclusive dressing for 24 hours. After removal of the dressing the skin was washed with lukewarm water. Animals were observed during a period of 14 days after removal of the dressing. No deaths occurred during the study. No clinical signs were observed. Due to a persisting blue staining of the skin a reliable assessment of potential skin irritating effects was not possible during the first study days. At later time points when the staining had vanished no reddening of the skin was seen. No gross findings were recorded during necropsy at the termination of the study at day 15. In conclusion, no adverse effects in rats were seen after the application of a single dermal dose of test material at a dose level of 2000 mg/kg bodyweight.