Pyrrolo[3,4-c]pyrrole-1,4-dione, 3,6-bis(4-chlorophenyl)-2,5-dihydro-

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 1986-01-20 to 1986-01-24

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Non-GLP guideline study according to OECD guideline 203 (limit test) without analytical monitoring

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1981

Deviations:

yes

Remarks:

limit test without stock solution

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION:
- Method: No stock solution was prepared. 100 mg/L test substance and 4 mg/L alkylbenzolesulphonate were premixed with water, then added directly to the water in the tank and homogeneously distributed to produce the desired test concentration.
- Controls: blank (dechlorinated tap water) and vehicle control
- Chemical name of vehicle: Alkylbenzenesulphonate
- Concentration of vehicle in test medium: 4 mg/L
- Evidence of undissolved material: yes, initially the test substance appeared homogeneously distributed at all test concentrations. A deposit was
observed after 30 minutes.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Length at study initiation: 22 mm (18 - 25 mm)
- Weight at study initiation: 0.09 g ( 0.06 - 0.11 g)
- Feeding during test: None
- Adaptation : 24 h
- Feeding: None 24 h prior to exposure

ACCLIMATION
- Acclimation period: 39 d

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

163 mg CaCO3/L

Test temperature:

23 ± 1 °C

pH:

8.0 - 8.1

Dissolved oxygen:

81 - 100 %

Nominal and measured concentrations:

Nominal: 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Aquaria
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: Glass (36/22/25 cm) 20 L filled with 15 L
- Aeration: None
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source of dilution water: Dechlorinated tap water (carbon filtered)

OTHER TEST CONDITIONS
- Light: Fluorescent, 16 h/d

EFFECT PARAMETERS MEASURED: Mortality, symptoms
TEST SYSTEM
- Test vessel: glass aquaria
- Type (delete if not applicable): not reported
- Material, size, headspace, fill volume: glass aquaria of 20 liters filled with 15 liters
- Aeration: none
- Type of flow-through (e.g. peristaltic or proportional diluter): not applicable
- Renewal rate of test solution (frequency/flow rate): not applicable
- No. of organisms per vessel: 10 or 20
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap water (carbon filter)

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16h
- Light intensity: not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality at 24, 48, 72 and 96h

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable as limit test
- Justification for using less concentrations than requested by guideline: limit test
- Range finding study: no
- Test concentrations: 100 mg/L (nominal)
- Results used to determine the conditions for the definitive study: not applicable

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No toxic effects occur within the range of solubility.

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No toxic effects occur within the range of solubility.

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No toxic effects occur within the range of solubility.

Details on results:

- Mortality of control and vehicle control: None
- Any observations that might cause a difference between measured and nominal values: Yes, at concentrations > 100 mg/L the test substance precipitated immediately

Validity criteria fulfilled:

yes

Conclusions:

In an acute fish toxicity study, the substance did not cause any mortality at the nominal limit test concetration of 100 mg/L.

Executive summary:

The purpose of this study was to evaluate the acute toxicity of the test substance on Zebrafish (Danio rerio) similar to OECD TG 203.

Fish were exposed to aqueous test media containing the test item at the single concentration of 100 mg/L (limit test)

for 96 hours. Concurrent negative and vehicle (alkylbenzenesulphonate) control groups were run.

A static test was performed by premixing 100 mg/L test substance and 4 mg/L of the vehicle alkylbenzolesulphonate and adding the mixture directly to the water in the tank. Test item precipitated immediately. Test item concentration were not measured analytically.

The fish were observed at approximately 24, 48, 72 and 96 hours for mortality.

All validity criteria were met. Mortality was not observed during the 96-hexposure period either in the treated or in the control groups.The 96-h NOEC and LC₀values were determinedto be 100 mg/L; the LC₅₀the LOEC, and LC₁₀₀values were determined to be > 100 mg/L.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 1986-01-20 to 1986-01-24

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Non-GLP guideline study according to OECD guideline 203 (limit test) without analytical monitoring

Justification for type of information:

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The read-across is performed between two forms of the same substance. The identities of the two forms are describe below.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
The source form is 3,6-bis(4-chlorophenyl)-2,5-dihydropyrrolo[3,4-c]pyrrol-1,4-dione (EC-no. 401-540-3), referred to here as PR254. PR254 is an organic mono-constituent substance with a typical purity of > 99.5% (w/w). It does not contain any impurity relevant for classification or labelling of the substance. The target form is the nanoform of the source substance, referred to here as PR254 nanoform. As the source form, it has a typical purity of > 99.5% (w/w) and it does not contain any impurity relevant for classification or labelling of the substance. The PR254 nanoform is spheroidal with a pure polyhedral shape and is not surface-treated.

3. ANALOGUE APPROACH JUSTIFICATION
The two analogue forms have the same structure. Under ambient atmosphere, the specific surface energy of particles increases with decreasing particle size. Therefore, particle aggregate to reach an energy minimum. The driving forces are hydrogen bonds and van der Waals forces (π-π interaction). Substantial energy is required to disperse the PR254 nanoform aggregates to particles that fall under the nanoform definition.
PR254 was been tested extensively addressing information requirements of Annexes VII to IX without identifying any biological target. PR254 nanoform could potentially have biological targets due to the different particle size distribution, which would require processes capable of dispersing the aggregates, e.g. in aqueous milieu. However, both forms have been tested according to OECD Test Guideline 318, demonstrating that PR254 nanoform cannot be dispersed under the condition of the study, i.e. immediately after sonification re-forms aggregates. Also, PR254 aggregates to a large extent, but can be more easily dispersed than the nanoform. The experiments demonstrated that exposure in aqueous milieu will be primarily to aggregates, regardless of the PR254 form.
Therefore, it is concluded that both forms will behave identically in studies, in which they are applied under atmospheric conditions and/or in aqueous milieus, so that for the PR254 nano-form no specific biological targets need to be considered.
As both forms form non-dispersible aggregates in aqueous milieu, read-across of aquatic toxicity studies from the source to the target form is scientifically justified.

4. DATA MATRIX
The data matrix is included as Annex 1 in the assessment report ‘PR254 bulk nano analogue approach 210111’ attached here below under ‘Attached justification’.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1981

Deviations:

yes

Remarks:

limit test without stock solution

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION:
- Method: No stock solution was prepared. 100 mg/L test substance and 4 mg/L alkylbenzolesulphonate were premixed with water, then added directly to the water in the tank and homogeneously distributed to produce the desired test concentration.
- Controls: blank (dechlorinated tap water) and vehicle control
- Chemical name of vehicle: Alkylbenzenesulphonate
- Concentration of vehicle in test medium: 4 mg/L
- Evidence of undissolved material: yes, initially the test substance appeared homogeneously distributed at all test concentrations. A deposit was
observed after 30 minutes.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Length at study initiation: 22 mm (18 - 25 mm)
- Weight at study initiation: 0.09 g ( 0.06 - 0.11 g)
- Feeding during test: None
- Adaptation : 24 h
- Feeding: None 24 h prior to exposure

ACCLIMATION
- Acclimation period: 39 d

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

163 mg CaCO3/L

Test temperature:

23 ± 1 °C

pH:

8.0 - 8.1

Dissolved oxygen:

81 - 100 %

Nominal and measured concentrations:

Nominal: 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Aquaria
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: Glass (36/22/25 cm) 20 L filled with 15 L
- Aeration: None
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source of dilution water: Dechlorinated tap water (carbon filtered)

OTHER TEST CONDITIONS
- Light: Fluorescent, 16 h/d

EFFECT PARAMETERS MEASURED: Mortality, symptoms
TEST SYSTEM
- Test vessel: glass aquaria
- Type (delete if not applicable): not reported
- Material, size, headspace, fill volume: glass aquaria of 20 liters filled with 15 liters
- Aeration: none
- Type of flow-through (e.g. peristaltic or proportional diluter): not applicable
- Renewal rate of test solution (frequency/flow rate): not applicable
- No. of organisms per vessel: 10 or 20
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap water (carbon filter)

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16h
- Light intensity: not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality at 24, 48, 72 and 96h

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable as limit test
- Justification for using less concentrations than requested by guideline: limit test
- Range finding study: no
- Test concentrations: 100 mg/L (nominal)
- Results used to determine the conditions for the definitive study: not applicable

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No toxic effects occur within the range of solubility.

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No toxic effects occur within the range of solubility.

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: No toxic effects occur within the range of solubility.

Details on results:

- Mortality of control and vehicle control: None
- Any observations that might cause a difference between measured and nominal values: Yes, at concentrations > 100 mg/L the test substance precipitated immediately

Validity criteria fulfilled:

yes

Conclusions:

In an acute fish toxicity study, the substance did not cause any mortality at the nominal limit test concetration of 100 mg/L.

Executive summary:

The purpose of this study was to evaluate the acute toxicity of the test substance on Zebrafish (Danio rerio) similar to OECD TG 203.

Fish were exposed to aqueous test media containing the test item at the single concentration of 100 mg/L (limit test)

for 96 hours. Concurrent negative and vehicle (alkylbenzenesulphonate) control groups were run.

A static test was performed by premixing 100 mg/L test substance and 4 mg/L of the vehicle alkylbenzolesulphonate and adding the mixture directly to the water in the tank. Test item precipitated immediately. Test item concentration were not measured analytically.

The fish were observed at approximately 24, 48, 72 and 96 hours for mortality.

All validity criteria were met. Mortality was not observed during the 96-hexposure period either in the treated or in the control groups.The 96-h NOEC and LC₀values were determinedto be 100 mg/L; the LC₅₀the LOEC, and LC₁₀₀values were determined to be > 100 mg/L.

Description of key information

The acute fish toxicity the substance was determined as LC50 >100 mg/L.

Additional information

As no mortality was observed in a limit test at 100 mg/L, no key value for risk assessment is reported.