bis(2-methylpropyl) perylene-dicarboxylate

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3,10-bis(2-methylpropyl) perylene-3,10-dicarboxylate; 3,9-bis(2-methylpropyl) perylene-3,9-dicarboxylate

EC Number:

616-749-7

Cas Number:

79869-59-3

Molecular formula:

C30H28O4

IUPAC Name:

3,10-bis(2-methylpropyl) perylene-3,10-dicarboxylate; 3,9-bis(2-methylpropyl) perylene-3,9-dicarboxylate

Details on test material:

Name of test substance: Oracet F Yellow 084
Test substance No.: 16/0121-1
Batch identification: 12-7612
CAS No.: 79869-59-3
Content: > 99.8 g/100 g
Identity: confirmed
Homogeneity: given
Storage stability: Expiry date: 26 Mar 2023
Chemical Name: Perylenedicarboxylic acid, bis(2-methylpropyl) ester
Date of production: unknown
Physical state: solid
Color: orange
Water solubility: <100 μg/L (pH at 23°C: 7.25)
Storage conditions: ambient

Sampling and analysis

Analytical monitoring:

no

Remarks:

Analyses of the test substance preparations was not conducted for technical reasons. No reliable method for analyses in the required concentration range could be developed.

Test solutions

Vehicle:

no

Details on test solutions:

The test substance is poorly water soluble; therefore a saturated stock solution was prepared following general guidance provided in OECD 23. Each test solution was prepared separately (differential loading) by directly adding test substance to test medium and stirring for about 2 days. After this time, undissolved test substance was removed by filtration with a membrane filter (pore width 0.2 μm). The first 50-100 mL of filtered solution was discarded (used to condition the filter). The exposure was started after separation of the undissolved material. The aqueous fraction of the test solution, after separation of the undissolved material, was considered 100% saturated in test media. According to OECD 23, test solutions of poorly soluble substances separated from the remaining undissolved fraction are considered equivalent to the nominal or loading concentration used to prepare them. Fresh test solutions were prepared and renewed daily to maximize the exposure to all soluble components of the test substance.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Test species: Daphnia magna STRAUS
Reason for selection of the test species: Recommended species in the test guidelines.
Origin: The clone of Daphnia magna STRAUS 1820 used was supplied by the Institut National de Recherche Chimique Appliquée, France, in 1978. From this date on this clone was cultured and bred continuously in the Ecotoxicology Laboratory of Experimental Toxicology and Ecology, BASF SE, Ludwigshafen Germany.
Culture conditions: Daphnia brood stock are kept in mass cultures consisting of approx. 20 – 30 parthenogenetically reproducing females for a maximum of 4 weeks and fed live unicellular algae (Desmodesmus subspicatus), cultured continuously at the test facility. All individuals in each mass culture originate from a single female and are thus genetic clones. After approximately 14 days the adults have produced at least 3 broods and the young can be used in tests. Offspring are removed from the mass cultures at least once daily during the normal work week to ensure that young daphnia are <24-h old (first instar) at start of the exposure. Detailed records are kept (in test facility archives) to monitor the health of Daphnia brood stock cultures including observations of young production, mortality, ephippia, and measurement of water chemistry parameters. Only young from healthy cultures without signs of stress are used for testing.
Acclimatization: The Daphnia are cultured under the identical conditions as the test including test media (Elendt M4), water quality, and temperature (20 ±1°C).
Age at start of exposure: <24 hours (at least 3rd brood progeny)

Reference substance testing:
In order to verify that the Daphnia magna culture is responding normally to toxic stress, tests with a reference substance, sodium chloride, are conducted monthly. Reference substance tests are conducted according to OECD 202 guidelines and in accordance with GLP, but without a GLP status. The EC50(48h) of the reference substance sodium chloride was 4.54 g/L (experiment date: 16 Aug 2016. This result is within the range of 3.88 – 7.22 g/L and indicates that the culture of Daphnia magna used in this study is responding normally to toxic stress.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

2.41 mmol/L

Test temperature:

19.8 – 20.7 °C (continuous monitoring)

pH:

7.9-8.0

Dissolved oxygen:

8.5-9.0 mg/L

Salinity:

n/a

Conductivity:

583 μS/cm

Nominal and measured concentrations:

nominal: 100 mg/L

Details on test conditions:

Test medium: A synthetic fresh water (Elendt M4) is used as media for culture and test purposes. For the composition of this M4 medium see OECD 202. The general properties of this medium are as follows.
Total hardness: 2.20 – 3.20 mmol/L
Acid capacity up to pH 4.3: 0.80 – 1.00 mmol/L
Molar ratio Ca:Mg: about 4 : 1
pH value: 7.5 – 8.5
Conductivity: 550 - 650 μS/cm
Total organic carbon: <2 mg/L
Dissolved oxygen: Must remain ≥3mg/L during the test. To assure optimal dissolved oxygen levels, the M4 medium is aerated for approximately 24 hours prior to use.
Test vessels: Numbered glass beakers (nominal volume 100 mL)
Test volume: 50 mL
Test solution renewal: Static-renewal exposure via test water.
Reason: In order to insure constant exposure conditions this study was conducted as a static-renewal exposure. The renewal period was 24 hours.
Test temperature: 19.8 – 20.7 °C (continuous monitoring)
Biological loading: 5 animals / test vessel (0.1 animals/mL)
Light intensity / Photo period: 161 – 677 lux at a wave length of 400-750 nm; 16 hours light : 8 hours darkness
Aeration: none
Diet: No feeding during the exposure period.
Test groups : 0 (control), 100 mg/L as nominal concentrations (loading concentration) based on test substance mass without correction for purity or composition
Replicates / test group: 4 replicates with test animals (5 animals per replicate)
Daphnids / test group: 20
Reason for the selection of the test concentrations: In a preliminary test (experimental conduct in accordance with
GLP, without a GLP status) a concentration of 100 mg/L caused no immobilisation or symptoms after 48 hours. According to the OECD-guideline, the highest suggested test concentration is 100 mg/L for a limit test. The raw data of the range finding test are archived together with the raw data of this study.

The exposure was started by impartially distributing 20 neonate (< 24 hours old) Daphnia magna evenly among the 4 test vessels per test group. The neonate daphnids all originated from the same mass culture and were thus genetic clones. The Daphnia were transferred into the test vessels with minimal culture water by capturing each daphnid in a pipette then gently expelling it onto a Teflon mesh (70 μm) held by forceps. Each daphnid was then immediately transferred from the mesh into the corresponding test vessel from lowest to highest concentration. Each test vessel was visually checked for immobilized daphnids after 0, 24 and 48 hours. In addition any abnormal behavior or appearance was documented. For test renewal, daphnids in each replicate were captured with a pipette and transferred into test vessels with fresh test solution. Throughout the test, the appearance of the test solutions and dissolution behavior of the test substance was observed and recorded daily in old and new test solutions. The chemical and physical parameters of the test medium (total hardness, acid capacity, pH, TOC and conductivity) were determined after aeration and prior to use in the test. The pH and dissolved oxygen content of the test solutions was measured after 0h and 24h (new test solutions) and after 24 h and 48 h (old test solutions) in replicate 1 of each tested concentration. In addition, temperature was measured continuously during the whole exposure period in a separate vessel filled with water proximal to the test vessels. The TOC (Total organic carbon) values of the test concentration and the control were analyzed at the start and the end of one renewal interval as an additional water quality parameter.

Reference substance (positive control):

yes

Remarks:

sodium chloride

Results and discussion

Effect concentrations

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Any other information on results incl. tables

All of the test solutions were visibly clear and colorless after preparation and remained so after 24 and 48 hours of exposure in the test. No undissolved test substance was visible and there were no other remarkable observations.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

This test was fully compliant with all the following validity criteria required by the corresponding test guidelines and is considered valid. • ≤10% immobilisation in the control • O2 concentration ≥3 mg/L in control and test vessels

Conclusions:

In a 48-hour acute toxicity study, neonate Daphnia magna were exposed to the test substance at nominal (loading) concentrations of 0 (control) and 100 mg/L under semi-static, daily-renewal conditions in accordance with the OECD 202 guideline. The water pH, temperature and dissolved oxygen were within acceptable guideline specifications. Immobilization (mortality) and sublethal effects were observed daily.

The 48 hour EC50 /EL50 values determined in this acute toxicity study were: >100 mg/L based on the nominal (loading) rate

The concentration of test substance in test media could not be verified analytically. According to guidance in OECD 23, for tests with chemicals that cannot be quantified by analytical methods at the concentrations causing effects, the effect concentration can be expressed based on the nominal concentrations. The toxicity results presented here are consistent with the results from preliminary tests. The results in this study are consistent with all validity criteria and the test is valid according to the guidelines of this study. No deviations from test guidelines or other incidents occurred during the course of the reported test which may have influenced the results.