Condensation products of Dihydrofuran-2,5-dione, mono-alkenyl derivs. with polyethylene amine and acetic anhydride

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 August 2013 to 31 October 2013

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- lot/batch No.of test material: E00031-743
- Purity: 100%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Dark storage place at room temperature
- Stability: Stable at normal temperature

Analytical monitoring:

yes

Remarks:

HPLC

Details on sampling:

The mixed solution was taken out with equal volume from the middle layer of each test vessel.

Vehicle:

no

Details on test solutions:

Due to the preliminay results, a limit test concentration of 100 mg/L was prepared by mixing a test sample with dilution water for 48 hours using a magnetic stirrer. Then, the suspension was filtered with a glass fibre filter by suction. The filtrate was stirred for 30 minutes to recover dissolved oxygen concentrations which were decreased by the suction filtration. The test solution was divided into each test vessel.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: The University of Sheffield, Sheffield, S10 2UQ, Uk)
- Age: Less than 24 hours old
- Feeding during test
Fed Chlorella vulgaris daily between 01. and 0.2 mg C/ Daphnia/ day

ACCLIMATION
Young daphnids produced by parents that had been cultured in the Kurume Laboratory were used. The parents to obtain young daphnids were bred in the same quality of water, water temperature and photoperiod as used in the test. The parental animals used for the test were the same lot and bred for more than 14 days.

Test type:

semi-static

Water media type:

other: ASTM

Limit test:

yes

Total exposure duration:

21 d

Post exposure observation period:

The animals were observed daily for mortality and the number of offspring produced until the end of the study.

Hardness:

150-180 mg/L CaCO3

Test temperature:

20 +/- 1 degrees Celsius

pH:

6 - 9

Dissolved oxygen:

8.5 - 8.9 mg/L

Nominal and measured concentrations:

Due to the poor water solubility, the dissolved concentration of the test item was below the determination limit of 0.227 mg/L, therefore the results are based on a nominal loading rate.

Details on test conditions:

TEST SYSTEM
- Test vessel: 80 mL/test vessel
- Aeration: no
- No. of organisms per vessel: 10 daphnids/test level
- No. of vessels per concentration (replicates): 10 replicates / test level

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water was used for dilution water. Selenium and Vitamin B12 were added at 1 microgram/litre.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light / 8 hours dark
- Light intensity: Artificial light of white fluorescence lamp not exceeding 15-20 uE.m-2.s1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and number of offspring were monitored daily. The appearance of the test solution was observed at the preparation, before renewal and once during the exposure period. Dissolved oxygen concentration, pH, water temperature, total hardness and light intensity were measured 6 times (triple repetitions weekly) during the exposure period for quality and once for the light intensity measurement.

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Duration:

21 d

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

ca. 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Details on results:

At the start of the exposure, the test solution was colourless and clear. The appearance before the renewal was light yellowish green by feeding chlorella. The temperature of the test solutions ranged from 20.2-20.5 degrees Celsius; the pH ranged from 8.1-8.4; the total hardness ranged from 17-164 mgCaC03/L; the light intensity was 15 uE.m^-2.s^-1. All of these results were within the appropriate environmental condition range.
The concentrations of the test item in the test solution used for exposure at the preparation stage, and before the renewal and the end of the exposure period, was not detectable (<0.227 mg/L). The measured concentrations of the test item in test solution used for exposure at the preparation ranged from 0.25-0.26 mg/L, and those before renewal or at the end of the exposure ranged from 0.45 mg/L to less than 0.227 mg/L.
The LC50 on both day 14 and day 21 were more than the solubility in dilution water. The LL50 for day 14 and day 21 were more than 100 mg/L nominal loading concentration. The EC50 and EL50 on both days 14 and 21 were more than the solubility in dilution water and more than 100 mg/L nominal loading concentration respectively. For 21-Day Daphnia reproduction, NOEC and LOEC were both expressed as “more than the solubility of dilution water” since no significant difference was obtained between the exposure group and the control.

Results with reference substance (positive control):

Not applicable

Validity criteria fulfilled:

yes

Conclusions:

No effects on the parental animals or on the reproduction were observed. Therefore, these results suggest that the test item has no effect on Daphnia reproduction and the NOEC on the reproduction is more than or equal to the solubility in dilution water.

Executive summary:

This study was conducted to confirm the reproductive effect of the test item on the test organisms around the solubility of the test item in dilution water. The test item was poorly soluble, below the determination limit (<0.227 mg/L). Therefore, the definitive study was carried out using saturated test solution with insoluble test item under a semi-static regime of three times renewal per week in consideration of maintaining the test item concentration in test solution. In the exposure period, increase in test item concentration was observed in the exposure level; the reason for this was unclear. No effects on the parental animals or on the reproduction were observed. Therefore, these results suggest that the test item has no effect on Daphnia reproduction and the NOEC on the reproduction is more than or equal to the solubility in dilution water. The environmental conditions were within the suitable range, moreover, it is concluded that this study complied with the applied test guidelines.

Description of key information

This study was conducted to GLP in accordance with the OECD 211 study.

No effects on the parental animals or on the reproduction were observed, therefore, these results suggest that the test item has no effect on Daphnia reproduction. The NOEC on the reproduction is more than or equal to the solubility in dilution water. The NOELR was 100 mg/kg, and the LL50 and EL50 were both more than 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L