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

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990-02-16 to 1990-08-09

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Modified GLP guideline study according to OECD Guideline 202 (Part II) without analytical monitoring

Qualifier:

according to guideline

Guideline:

other: OECD-Guideline No.: 202, Part II, 1984 semi-static system, modified according to the draft guideline arising out of the EEC/OECD ringtest 1985-1986

Deviations:

yes

Remarks:

, see "Principles of method if other than guideline"

Principles of method if other than guideline:

- Test concentration: Testwater saturated with test substance
- According to EEC/OECD ringtest (1987): 10 animals per concentration and control 1 animal per vessel.
- 5 additional vessels to account for the occurence of males and/or injured daphnids (day 0 to 13)
- Measurement of oxygen in 1 vessel per concentration

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 2.0 g of the test substance were mixed with and made up to 2000 mL of testwater. This slurry was continously agitated during 48 hours at room temperature and finally filtered (Millipore filter 0.22 µm) before use. This solution without further dilution was used for the exposure of the daphnids. The stock solution was directly used after filtration. The saturated solution was clear and colorless from start of exposure to renewal.
- Controls: water
- Evidence of undissolved material: None, the saturated solution was clear and colorless from start of exposure to renewal

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Source: CIBA-GEIGY Ltd., Facilities, Basel, Switzerland
- Age at study initiation: 0 - 24 h
- Method of breeding: Cultures of daphnids are maintained in glass vessels containing approximately 3L of reconstituted water (according to guideline) at 20 ± 1 °C (water is renewed partially thrice weekly). At each renewal the adult daphnids are fed with a suspension of green algae (Scenedesmus subspicatus) 1.25 mL of approx. 1E6 cells/mL per liter water supplemented by 1.5 ml suspension of TETRAMIN-extract per liter water.
- Pretreatment: 24 hours before the start of the test reproductive daphnia are separated from the young by sieving through a 800 micrometer sieve. This operation is repeated immediately before the start and the young daphnia (0 - 24 h old) are retained for the test.
- Feeding during test: Daily (Monday through Saturday)

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Hardness:

240 mg/L CaCO3

Test temperature:

20 ± 1 °C

pH:

7.9 - 8.8

Dissolved oxygen:

85 - 121 °C

Nominal and measured concentrations:

Only one concentration tested (saturated solution)

Details on test conditions:

TEST SYSTEM
- Test vessel: Beakers covered with watch glasses; 50 mL solution per beaker; 10 beakers per concentration with 1 daphnia each; 5 additional vessels to account for the occurence of males and/or injured daphnids (day 0 to 13)
- Aeration: None
- Renewal rate of test solution: Renewal of test solutions on Mondays, Wednesdays and on Fridays.
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Intervals of water quality measurement: pH was measured daily (Monday - Saturday) before and after each renewal of the test solutions; temperature was measured continuously; dissolved oxygen was measured daily (Monday - Saturday) immediately before the renewal of the test solutions

OTHER TEST CONDITIONS
- Photoperiod: 16h daily, approx. 1500 lux

EFFECT PARAMETERS MEASURED: Reproduction, mobility

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

NOEC

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

reproduction

Remarks on result:

other: No effects occur within the range of solubility.

Details on results:

- Mortality of parent animals: None
- No. of offspring produced per female: 42 (blank); 46 (saturated solution 1); 54 (sat. solution 2)

Validity criteria fulfilled:

yes

Conclusions:

In a long-term (21 days) semi-static daphnia reproduction study, the substance did not cause any effects using a saturated solution.

Executive summary:

The long-term effect of the substance on daphnia magna reproduction was investigated in a GLP-study similar to OECD TG 202 exposing daphnia for 21 days.

Due to the very low water solubility, a saturated solution was tested under semi-static conditions (without analytical measurements).

Per control and treatment, 10 daphnia were used per treatment and control.

The test substance did not induce any immobilisation, nor did it affect the reproduction.

Reference 2

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990-02-16 to 1990-08-09

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Modified GLP guideline study according to OECD Guideline 202 (Part II) 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:

other: OECD-Guideline No.: 202, Part II, 1984 semi-static system, modified according to the draft guideline arising out of the EEC/OECD ringtest 1985-1986

Deviations:

yes

Remarks:

, see "Principles of method if other than guideline"

Principles of method if other than guideline:

- Test concentration: Testwater saturated with test substance
- According to EEC/OECD ringtest (1987): 10 animals per concentration and control 1 animal per vessel.
- 5 additional vessels to account for the occurence of males and/or injured daphnids (day 0 to 13)
- Measurement of oxygen in 1 vessel per concentration

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 2.0 g of the test substance were mixed with and made up to 2000 mL of testwater. This slurry was continously agitated during 48 hours at room temperature and finally filtered (Millipore filter 0.22 µm) before use. This solution without further dilution was used for the exposure of the daphnids. The stock solution was directly used after filtration. The saturated solution was clear and colorless from start of exposure to renewal.
- Controls: water
- Evidence of undissolved material: None, the saturated solution was clear and colorless from start of exposure to renewal

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Source: CIBA-GEIGY Ltd., Facilities, Basel, Switzerland
- Age at study initiation: 0 - 24 h
- Method of breeding: Cultures of daphnids are maintained in glass vessels containing approximately 3L of reconstituted water (according to guideline) at 20 ± 1 °C (water is renewed partially thrice weekly). At each renewal the adult daphnids are fed with a suspension of green algae (Scenedesmus subspicatus) 1.25 mL of approx. 1E6 cells/mL per liter water supplemented by 1.5 ml suspension of TETRAMIN-extract per liter water.
- Pretreatment: 24 hours before the start of the test reproductive daphnia are separated from the young by sieving through a 800 micrometer sieve. This operation is repeated immediately before the start and the young daphnia (0 - 24 h old) are retained for the test.
- Feeding during test: Daily (Monday through Saturday)

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Hardness:

240 mg/L CaCO3

Test temperature:

20 ± 1 °C

pH:

7.9 - 8.8

Dissolved oxygen:

85 - 121 °C

Nominal and measured concentrations:

Only one concentration tested (saturated solution)

Details on test conditions:

TEST SYSTEM
- Test vessel: Beakers covered with watch glasses; 50 mL solution per beaker; 10 beakers per concentration with 1 daphnia each; 5 additional vessels to account for the occurence of males and/or injured daphnids (day 0 to 13)
- Aeration: None
- Renewal rate of test solution: Renewal of test solutions on Mondays, Wednesdays and on Fridays.
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Intervals of water quality measurement: pH was measured daily (Monday - Saturday) before and after each renewal of the test solutions; temperature was measured continuously; dissolved oxygen was measured daily (Monday - Saturday) immediately before the renewal of the test solutions

OTHER TEST CONDITIONS
- Photoperiod: 16h daily, approx. 1500 lux

EFFECT PARAMETERS MEASURED: Reproduction, mobility

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

NOEC

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

reproduction

Remarks on result:

other: No effects occur within the range of solubility.

Details on results:

- Mortality of parent animals: None
- No. of offspring produced per female: 42 (blank); 46 (saturated solution 1); 54 (sat. solution 2)

Validity criteria fulfilled:

yes

Conclusions:

In a long-term (21 days) semi-static daphnia reproduction study, the substance did not cause any effects using a saturated solution.

Executive summary:

The long-term effect of the substance on daphnia magna reproduction was investigated in a GLP-study similar to OECD TG 202 exposing daphnia for 21 days.

Due to the very low water solubility, a saturated solution was tested under semi-static conditions (without analytical measurements).

Per control and treatment, 10 daphnia were used per treatment and control.

The test substance did not induce any immobilisation, nor did it affect the reproduction.

Description of key information

In a long-term (21 days) semi-static daphnia reproduction study, the substance did not cause any effects using a saturated solution.

Additional information

As the substance did not induce any effects at its maximum water solubility, no key value is indicated.

Note also that a long-term toxicity study with aquatic invertebrates would not have been required, if it had not existed already, as the hazard assessment revealed neither a need to classify the substance as dangerous to the environment, nor that it is a PBT or vPvB substance, nor that there are any further indications that the substance may be hazardous to the environment (see Annex I of Regulation (EC) No 1907/2006).