Reaction mass of 3,6-Bis(3-chlorophenyl)-2,5-dihydro-pyrrolo[3,4-c]pyrrole-1,4-dione, 3-(3-Chlorophenyl)-6-(4-chlorophenyl)-2,5-dihydro- pyrrolo[3,4-c]pyrrole-1,4-dione and 3,6-Bis(4-chlorophenyl)-2,5-dihydro-pyrrolo[3,4-c]pyrrole-1,4-dione

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147

GLP compliance:

yes

Test material

Test material information

Constituent 1

Details on test material:

- Description: Orange-red powder
- Test substance storage: At room temperature in the dark
- Stability under storage conditions: Stable
- Expiry date: 15 August 2007

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charies River Deutschland, Kisslegg, Germany
- Age at study initiation: 7 - 14 weeks
- Weight at study initiation: 1458 - 2552 g
- Housing: Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5 cm).
- Diet: Standard laboratory rabbit diet (Charies River Breeding and Maintenance Diet for Rabbits, Altromin, Lage, Germany) approx. 100 g. per day. Hay (TecniLab-BMl BV, Someren, The Netherlands) was provided at least three times a week.
- Water: Free access to tap water.
- Acclimation period: Acclimatisation period was at least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.0- 21.8
- Humidity (%): 35-65
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

other: watery ethanol (50% v/v) (water; Milli-U, Millipore Corporation, Bedford, USA; ethanol, Merck, Darmstadt, Germany)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5g of the test substance. The test substance was moistened with 1 mL of the vehicle


VEHICLE
- Amount(s) applied (volume or weight with unit): 1 mL
- Concentration (if solution): watery ethanol (50% v/v) (water; Milli-U, Millipore Corporation, Bedford, USA; ethanol, Merck, Darmstadt, Germany)

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

STUDY DESIGN
The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 7 days later, after considering the degree of skin irritation observed in the first animal.

TREATMENT
Approximately 24 hours before treatment, the dorsal fur was clipped with electric clippers, exposing an area of approximately 150 square centimeters (10x15 cm²). To facilitate scoring, treated skin areas were re-clipped at least 3 hours before the observations. Each animal was treated by dermal application of 0.5g of the test substance. The test substance was moistened with 1 mL of the vehicle and applied to the skin of one flank, using a metalline patch of 2x3 cm. The patch was mounted on Micropore tape, which was wrapped around the abdomen and secured with Coban elastic bandage. Four hours after the application, the dressing was removed and the skin cleaned of residual test substance using tap water, watery ethanol (50% v/v) (water; Milli-U, Millipore Corporation, Milli-U, Millipore Corporation, Bedford, USA; acetone, Labscan, Dublin, Ireland). For one animal the skin was also cleaned on day 2 with watery acetone (50% v/v) (tap water; acetone, Labscan, Dublin, Ireland). The treated skins were still stained by the test substance, but erythema could be assessed from 48 hours after application of the test substance onwards. After the final observation, the animals were sacrificed by intra-venous injection of pentobarbital (Euthesate®; Sanofi Sante B.V., Maassluis, The Netherlands).

OBSERVATIONS
Mortality/Viability: Twice daily; Toxicity: At least once daily; Body Weight: Day of treatment (prior to application) and at termination; Irritation: The skin reactions were assessed at approximately 1, 24,48 and 72 hours after the removal of the dressings and test substance. The irritation scores and a description of all other (local) effects were recorded. Adjacent areas of the untreated skin of each animal served as controls.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 48 - 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: Orange staining of the skin by the test substance, therfore at the 1 and 24 hours time point no scoring possible.

Results 2

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 1 - 72 hours

Score:

0

Max. score:

4

Irritant / corrosive response data:

No skin irritation was caused by 4 hours exposure to the test substance. The observed mean values (24 - 72 h or 48 - 72 h) of individual animals for edema and erythema were 0 without exception.

Other effects:

Colouration / Remnants: Orange staining of the treated skin by the test substance was observed throughout the observation period. This hampered the skin irritation assessment on day 1 and 2 only.
Toxicity / Mortality: No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met