(4aR*, 8aR*)-1-bromo-4a,5,9,10,11,12-hexahydro-3-methoxy-6H-benzofuro[3a,3,2-ef] [2]benzazepin-6-one hydrochloride (1:1)

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2001-10-08 to 2002-03-07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 3.4, 7.5, 15, 34 and 75 mg/L (nominal)
- Sampling method: Duplicate samples of the test medium and controls were taken just before the start of the experiment and after 48 hours.
- Sample storage conditions before analysis: All samples were deep-frozen (at about -20 deg C) immediately after sampling. Based on pre-experiments for investigation of the storage stability, the test item is sufficiently stable in the test water under these storage conditions.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test media were prepared just before introduction of the daphnids. The test medium of the highest test concentration of nominal 75 mg/L was prepared by dissolving 45 mg test item completely in 600 mL test water by ultrasonic treatment for 10 minutes and intense stirring for 10 minutes at room temperature. Adequate volumes of this test medium were diluted with test water to prepare the test media with the lower test item concentrations.
- Eluate: no data
- Differential loading: no data
- Controls: yes, reconstituted test water
- Chemical name of vehicle: not applicable
- Concentration of vehicle in test medium: not applicable
- Evidence of undissolved material: no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain: Daphnia magna Straus.
- Source: A clone of this species was originally supplied by the University of Sheffield/UK in 1992, defined from the supplier as clone 5.
- Age at study initiation: 6-24 hours
- Weight at study initiation: no data
- Length at study initiation: no data
- Valve height at study initiation, for shell deposition study: not applicable
- Peripheral shell growth removed prior to test initiation: not applicable
- Method of breeding: A clone of this species was originally supplied by the University of Sheffield/UK in 1992, defined from the supplier as clone 5. Since this date, the clone is bred in the laboratories of RCC in reconstituted water of the quality identical to the water quality used in the tests (regarding pH, main ions and total hardness) and under temperature and light conditions identical to those of the tests.
- Feeding during test
- Food type: not applicable
- Amount: not applicable
- Frequency: not applicable

ACCLIMATION
Bred in water quality and conditions identical to those of the tests.
- Acclimation period: not applicable
- Acclimation conditions: not applicable
- Type and amount of food: not applicable
- Feeding frequency: not applicable
- Health during acclimation: not applicable

QUARANTINE (wild caught)
- Duration: not applicable
- Health/mortality: not applicable

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

water hardness: 2.5 mmol/L (250.0 mg/L) as CaCO3

Test temperature:

21 deg C

pH:

7.9 - 7.6

Dissolved oxygen:

8.2 (0 hours) - 7.9 (48 hours) mg/L

Salinity:

no data

Nominal and measured concentrations:

nominal concentrations: 3.4, 7.5, 15, 34 and 75 mg/L
measured concentrations: not analysed, not analysed, 12, 26, and 58 mg/L
The measured test substance concentrations were in the range 88 to 97% of the nominal concentrations. The lowest two concentrations were not analyzed because they were below the 48-h NOEC.

The measured test item concentrations in the analyzed test media samples were in the range of 88 to 97% of the nominal at the start of the test. This shows the correct dosage of the test concentrations. The concentrations of T002102 decreased to values in the range from 62 to 65% of the nominal values after the 48-hour test period. The losses may be due to instability of the test item in test water. The HPLC chromatograms of the aged treatment samples show a new peak
with a shorter retention time.
Therefore, all biological results are related to the mean measured test item concentrations, calculated as the average over all measurements per test concentration during the test period.

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL glass beakers
- Type (delete if not applicable): closed; The beakers were covered with glass plates to reduce the loss of water and to avoid the entry of dust into the solutions.
- Material, size, headspace, fill volume: 50 mL fill volume
- Aeration: The test water was aerated prior to the start of the study until oxygen saturation was reached. During the test period, the test water was not aerated.
- Type of flow-through: not applicable
- Renewal rate of test solution: not applicable
- No. of organisms per vessel: 10
- No. of vessels per concentration: 2
- No. of vessels per control: 2
- No. of vessels per vehicle control: not applicable
- Biomass loading rate: less than one daphnia per 2 mL

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted test water: analytical grade salts were dissolved in purified water to obtain the following nominal concentrations:
CaCI2 x 2H2O 2.0 mmol/L (294.0 mg/L)
MgSO4 X 7H2O 0.5 mmol/L (123.0 mg/L)
NaHCO30.75 mmol/L (65.0 mg/L)
KCI 0.075 mmol/L (5.8 mg/L)

- Total organic carbon: no data
- Particulate matter: no data
- Metals: no data
- Pesticides: no data
- Chlorine: no data
- Alkalinity: reconstituted test water = 0.8 mmol/L
- Ca/Mg ratio: reconstituted test water = 4/1
- Conductivity: no data
- Culture medium different from test medium: no
- Intervals of water quality measurement: no data

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: a 16-hour light to 8-hour darkness photoperiod (with a 30-minute transition period)
- Light intensity: between 370 and 420 Lux

EFFECT PARAMETERS MEASURED:
Immobility of the daphnids was determined by visual controls after 24 and 48 hours of exposure.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: no data
- Justification for using less concentrations than requested by guideline: not applicable
- Range finding study: yes
- Test concentrations: no data
- Results used to determine the conditions for the definitive study: yes, the test concentrations were based on the results of a range-finding test and on results of a pre-experiment to the solubility of the test item.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

31 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL 22-44 mg/L

Details on results:

- Behavioural abnormalities: After 48 hours of exposure, no immobility of the test organisms or visible abnormalities were observed in Daphnia magna exposed to concentrations of up to and including 12 mg/L. At the concentration of 26 mg/L, 30% of the daphnids were found to be immobile. At the highest test concentration of 58 mg/L, all daphnids were immobile.
- Observations on body length and weight: no data
- Other biological observations: none
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No remarkable observations were made concerning the appearance of the test media. All test media were clear solutions throughout the entire test duration.
- Effect concentrations exceeding solubility of substance in test medium: no
- 48-hour EC0 = 12 mg/L
- 48-hour NOEC = 12 mg/L.
- 48-hour EC100 = 58 mg/L.

Reported statistics and error estimates:

The 48-hour EC50 and the 95% confidence limits were calculated by moving average interpolation. All reported biological results are based on the mean measured test item concentrations. The lowest test concentrations of nominal 3.4 and 7.5 mg/L were not taken into account at the calculation because they were below the determined 48-hour NOEC and thus not analyzed.

Validity criteria fulfilled:

yes

Conclusions:

A 48-h immobilization test with the aquatic invertebrate Daphnia magna resulted in a measured 48-h EC50 of 31 mg/L, with 95% confidence limits from 22 to 44 mg/L. The result of the test can be considered reliable without restriction.

Description of key information

In the guideline study (Piether, 2002) 48-h immobilization test was carried out with the aquatic invertebrate Daphnia magna with T002102. The measured 48-h EC50 of 31 mg/L, with 95% confidence limits from 22 to 44 mg/L was obtained. The results of the test can be considered reliable without restriction.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

31 mg/L