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

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Substance type: pigment
- Physical state: solid
- Purity test date: Feb 13, 2013
- Expiration date of the lot/batch: Oct 2014
- Stability under test conditions: stable
- Storage condition of test material: at room temperature

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories B.V., The Netherlands
- Age at study initiation: 8 to 9 weeks
- Weight at study initiation: Males: 256.8 to 285.6 g, Females: 170.5 to 219.3 g
- Fasting period before study: no
- Housing: groups of 5 of the same sex
- Diet: ad libitum
- Water: e.g. ad libitum
- Acclimation period: 5 - 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C,
- Humidity (%): 30 - 70%
- Photoperiod (hrs dark / hrs light): hour fluorescent light / 12 hour dark cycle.

IN-LIFE DATES: From: Jan 31, 2013 To: March 27, 2013

Administration / exposure

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: flow-past, nose-only exposure system
- Method of holding animals in test chamber: restraint tubes
- Source and rate of air: 1.0 L/min
- System of generating particulates/aerosols: A dust aerosol was generated from the test item using a CR3020 rotating brush aerosol generator
connected to an AirVac TD110M. The aerosol generated was then discharged into the exposure chamber through a 63Ni charge neutralizer.
- Method of particle size determination: Mercer 7 stage cascade Impactor
- Temperature, humidity, pressure in air chamber: The temperature and relative humidity of the test atmosphere was measured continuously during
exposure using a calibrated device. The results were recorded manually and are reported in 30 minute intervals from the start of exposure.

TEST ATMOSPHERE
- Brief description of analytical method used: Gravimetric determination of aerosol concentration was performed twice (group 1) or six times
(groups 2 and 3) during exposure. The samples were collected on a Millipore®durapore filter, Type HVLP loaded in a 47 mm in-line stainless steel filter sampling device. The filters were weighed before and immediately after sampling using a calibrated balance. The test aerosol concentration was calculated from the amount of test item present on the filter and the sample volume.
- Samples taken from breathing zone: yes

TEST ATMOSPHERE : see table

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: Limit dose

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

0.5, 1 and 5 mg/L

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of weighing: Weighing on test days 1 (before exposure), 2, 4, 8 and 15 (before necropsy).
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: Observations for viability were recorded once before exposure on the day of exposure (test day 1), three times during exposure, immediately and 1 h after exposure on test day 1 and twice daily during the observation period. Each animal was examined three times during exposure, immediately and 1 h after exposure on test day 1 and once daily during the observation period. Observations were detailed and carefully recorded using explicitly defined scales as appropriate. Only grossly abnormal signs were detectable during exposure as the animals were restrained in the exposure tubes.

Statistics:

No statistical analysis was performed.

Results and discussion

Effect levels

Effect levels 1

Sex:

male/female

Dose descriptor:

LC0

Effect level:

1 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Effect levels 2

Sex:

male/female

Dose descriptor:

LC100

Effect level:

5 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

All animals exposed to 5.0 mg/l air died within two hours after exposure start. All animals exposed to 0.52 or 1.0 mg/L air survived the scheduled observation period.

Clinical signs:

other: In one male and two females exposed to 5.0 mg/L air, clinical signs such as increased activity, tachypnea and reduced body temperature were recorded before their death during exposure; the remaining seven animals exposed to 5.0 mg/L air died during exposu

Body weight:

Between test days 1 and 2, slight body weight loss was noted in all animals exposed to
0.52 mg/L or 1.0 mg/L air. This effect persisted in two females exposed to 0.5 mg/L air as well
as in two females exposed to 1.0 mg/L air and in up to day 4 of treatment. From test day 4
onwards, these females showed normal body weight gain. In the remaining animals exposed to
0.52 or 1.0 mg/L air, normal body weight development was noted from day 2 of treatment
onwards.

Gross pathology:

After their spontaneous death during exposure, all animals exposed to 5.0 mg/L air showed reddish discolored and incompletely collapsed lungs at necropsy. Although a relation to the treatment with test item cannot be fully excluded (the substance is an orange pigment) - this finding is considered to be most likely
secondary to the spontaneous death of the animals. In animals exposed to 0.52 or 1.0 mg/L air there were no macroscopic findings at scheduled necropsy.

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria