Reaction mass of 1,1'-(isopropylidene)bis[3,5-dibromo-4-(2,3-dibromo-2-methylpropoxy)benzene] and 1,3-dibromo-2-(2,3-dibromo-2-methylpropoxy)-5-{2-[3,5-dibromo-4-(2,3,3-tribromo-2-methylpropoxy)phenyl]propan-2-yl}benzene

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:

16 Aug - 05 Nov 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: the control and the prepared saturated solution of the test item were sampled
- Sampling method: 2 mL test samples were taken from one replicate vessel at each test concentration by fresh preparation of the test solution and prior to renewal
- Sample storage conditions before analysis: stored in the refrigerator (approx. 5 °C) if not analyzed within 24 h

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: a nominal concentration of 10 mg/L of the test substance was prepared and stirred in the dark (72 h, 500 rpm) followed by filtration using a 0.22 µm nitrocellulose membrane to make a saturated solution

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: MicroBio Tests Inc.
- Age of parental stock: female daphnia (the parent animals), aged less than 24 h at the start of the test
- Feeding during test: yes
- Food type: concentrated freshwater suspensions of unicellular green algae Desmodesmus subspicatus and Pseudokirchneriel/a subcapitata
- Amount: rate equivalent to 0.1-0.2 mg of organic carbon per adult per day

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Hardness:

160 - 185 mg/L (as CaCO3)

Test temperature:

20.1 - 21.1 °C

pH:

7.388 - 8.667

Dissolved oxygen:

8.12 - 9.76 mg/L

Nominal and measured concentrations:

- control, 10 mg/L (nominal)
- control and treatment < LOD = 89.6 ng/L (measured)

Details on test conditions:

TEST SYSTEM
- Test vessel: 75 mL glass containers
- Fill volume: 50 - 60 mL
- Aeration: no
- Feeding during test: fed concentrated freshwater suspensions of unicellular green algae Desmodesmus subspicatus and Pseudokirchneriel/a subcapitata at a rate equivalent to 0.1-0.2 mg of organic carbon per adult per day
- Renewal rate of test solution (frequency/flow rate): 3 times per week renewal (parent animals were transferred by a glass pipette of suitable diameter)
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium contained: Stock solution I (H3BO3 2859.5 µg/L, MnCl2 x 4 H2O 360.5 µg /L, LiCI 306 µg/L, RbCI 71 µg/L, SrCl2 x 6 H2O 152 µg/L, NaBr 16 µg/L, Na2MoO4 x 2 H2O 63 µg/L, CuCl2 x 2 H2O 16.75 µg/L, ZnCl2 13 µg/L, CoCl2 x 6 H2O 10 µg/L, Kl 3.25 µg/L, Na2SeO3 2.19 µg/L, NH4VO3 0.575 µg/L, Na2EDTA x 2 H2O 250 µg/L, FeSO4 x 7 H2O 99.55 µg/L); Stock solution II (CaCI2 x 2 H2O 293.8 mg/L, MgSO4 x 7 H2O 123.3 mg/L, KCI 5.8 mg/L, NaHCO3 64.8 mg/L, Na2SiO3 x 9 H2O 10 mg/L, NaNO3 274 µg/L, KH2PO4 143 µg/L, K2HPO4 184 µg/L); Vitamin stock solution (Thiamine hydrochloride 75 µg/L, Cyanocobalamine (812) 1.0 µg/L, Biotine 0.75 µg/L). Medium was prepared by analytical reagent and deionized wate and aerated for 30 min prior to the start of the test and placed in testing temperature at least 12 h before the test.
- Total organic carbon: < 0.4 mg/L (before addition of algae)
- Culture medium different from test medium: no
- Intervals of water quality measurement: oxygen concentration, temperature, hardness and pH were measured twice per week, in fresh and old medium, in the control and in the treatment group

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light / 8 h dark
- Light intensity: 1000 - 1500 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : offspring produced by each parent animal were removed and counted from the appearance of the first brood. The numbers of living offspring were counted and the presence of aborted eggs or dead offspring were recorded daily

TEST CONCENTRATIONS
- Range finding study: yes, 4 replicates with 1 animal each were tested per test concentration
- Test concentrations: 1, 10 and 100% saturated solution (semi static test, 3 renewals / week). 51.3 mg test substance were added to glass containers with 5 L Elendt M4 medium and solutions were stirred 72 h at 500 rpm followed by a filtration through 0.221 µm membrane filter. This solution was used as the highest concentration. The other concentration were prepared by diluting in Elendt M4 medium.
- Results used to determine the conditions for the definitive study: based on the result of the range-finding test, a limit test with a 100% saturated solution was carried out

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 10 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test material (saturated solution)

Basis for effect:

reproduction

Remarks on result:

other: Measured concentrations of the test substance were below the limit of detection (89.6 ng/mL)

Details on results:

- Mortality of control: 0%
- The measured concentrations of the test substance were lower than the limit of detection of analytical method (LOD = 89.6 ng/mL).

Reported statistics and error estimates:

The differences of the average number of the offsprings during 21 d test between the control group and the treatment group were determined by using the Homoscedastic t Test. The data analyses was carried out with Toxcalc software (v 5.0.26).

- The NOEC (21 d) value was higher than or equal to the concentration of the saturated solution (which the measured concentration was lower than LOD = 89.6 ng/mL)

- The LOEC (21 d) value was higher than the concentration of the saturated solution (measured concentration was lower than LOD = 89.6 ng/mL)

- The EC50 (21 d) value was higher than the concentration of the saturated solution (measured concentration was lower than LOD = 89.6 ng/mL)

- The mortality of the parent animals in the control and in the treatment was 0% at the end of the test.

- The mean number of live offspring produced per parent animal surviving at the end of the test in the control was 122 and in the treatment 115.

Description of key information

NOEC (21 d) ≥ 10 mg/L (nominal) for Daphnia magna (OECD 211)

Additional information

The long term influence of Reaction mass of 1,1'-(isopropylidene)bis[3,5-dibromo-4-(2,3-dibromo-2-methylpropoxy)benzene] and 1,3-dibromo-2-(2,3-dibromo-2-methylpropoxy)-5-{2-[3,5-dibromo-4-(2,3,3-tribromo-2-methylpropoxy)phenyl]propan-2-yl}benzene to aquatic invertebrates was tested in a 21 d reproduction test according to OECD 211 (Jing, 2014). A single nominal concentration of 10 mg/L of the test substance was prepared stirred and filtered in order to prepare a saturated solution for the test, as the test substance is poorly soluble in water. The reproduction of the test organism Daphnia magna was monitored in a semi static system. The test solution was renewed 3 times per week. The concentrations of the test substance in the test solutions were analyzed by UPLC-MS. The measured concentrations of the test substance were lower than the limit of detection of (89.6 ng/mL). Under the testing conditions the reproduction of Daphnia magna was not affected by the test substance and therefore a NOEC (21 d) of ≥ 10 mg/L (nominal) was determined.