6H-Benzofuro[3a,3,2-ef][2]benzazepin-6-one, 1-bromo-4a,5,9,10,11,12-hexahydro-3-methoxy-11-methyl-, (4aR,8aR)-rel-

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2001-11-07 to 2002-02-26

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

439-750-2

EC Name:

-

Cas Number:

183487-56-1

Molecular formula:

Hill formula: C17H19BrClNO3 CAS formula: C17H19BrClNO3

IUPAC Name:

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

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Physical state: pale yellow solid (powder)
- Source and lot/batch No.of test material: RT002113G3A231
- Expiration date of the lot/batch: 2001-12-01 (retest date)
- Purity test date: no data


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature at about 20°C away from sunlight
- Stability under test conditions: no data
- Solubility and stability of the test substance in the solvent/vehicle: no data

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: duplicate samples were taken from the test media. The samples were acidified to about pH 3-5 immediately after sampling using 0.1 M phosphoric acid solution.
- Sample storage conditions before analysis: samples were analyzed immediately after sampling without prior storage.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: a concentrated stock solution of nominal 100 mg/L is prepared at the start of the test by dissolving 1000 mg T002113 in 10 L test water by intense stirring for 5 minutes at room temperature. The lower test item concentrations were prepared by diluting the highest test concentration. The test media were freshly prepared just before introduction of the fish.
- Eluate: no
- Controls: yes

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Source: The test fish were obtained from Aquarium Center Wildenauer, Munich, Germany.
- Length at study initiation (length definition, mean, range and SD): 3.1 ± 0.3 cm
- Weight at study initiation (mean and range, SD): 0.32 ±0.12 g
- Feeding during test: no
- Food type: commercial diet fish (Tetra min Hauptfutter)

ACCLIMATION
- Acclimation period: one week prior the start to the same temperature and test water
- Acclimation conditions (same as test or not): yes
- Type and amount of food: commercial diet fish (Tetra min Hauptfutter)
- Health during acclimation (any mortality observed): no fish died and all fish were healthy

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

125 mg CaCO3/L

Test temperature:

22-23°C

pH:

7.3-7.9

Dissolved oxygen:

at least 8.0 mg/L or higher

Salinity:

not applicable

Conductivity:

not indicated

Nominal and measured concentrations:

nominal test concentrations: 4.6, 10, 22, 46, 100 mg/L
mean measured concentrations at the start of the test: 4.87, 10.4, 22.9, 46.1, 93.4 mg/L
mean measured concentrations after 2 days: 2.96, 6.49, 14.0, not applicable, not applicable mg/L
mean measured concentrations after 4 days: 2.88, 6.36, not applicable, not applicable, not applicable mg/L.

The mean measured concentrations (calculated as the average over all measurements per test concentration) ranged from 84 to 77% of nominal.

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquarium
- Material, size, headspace, fill volume: glass aquarium with 5 liter test medium
- Aeration: slightly aerated
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): not reported
- No. of vessels per control (replicates): not reported
- No. of vessels per vehicle control (replicates): not reported
- Biomass loading rate: lower than 1 g fish wet weight per liter test medium.


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionized water
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4 : 1 (base on molarity)
- Intervals of water quality measurement: daily

OTHER TEST CONDITIONS
- Photoperiod: 16-hour light to 8-hour darkness with a 30 minute transition period
- Light intensity: 50 to 500 lux

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 to 2.2
- Range finding study
- Test concentrations: data not shown
- Results used to determine the conditions for the definitive study: yes, range-finding test (without GLP)

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

12 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 7.8-19

Details on results:

- Behavioural abnormalities: apathy
- Mortality of control: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: foaming
- Effect concentrations exceeding solubility of substance in test medium: no remarkable observation, clear test medium.
- 96-h NOEC = 7.8 mg/L; 96-h LC100 = 19 mg/L

Reported statistics and error estimates:

- The LC50 and the 95% confidence interval at the observation dates could not be calculated by Probit Analysis or Moving Average Interpolation due to the steep concentration-effect relationship. Instead the LC50-values were determined as the geometric mean value of the two consecutive test concentrations with 0 and 100% mortality, and the 95% confidence intervals for the LC50 as the test concentrations with 0 and 100% mortality. The calculations of the LC values are based on the mean measured test item concentrations.
- The NOEC, LOEC, LCO and LC100 were determined directly from the raw data.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of T002113 to zebra fish was determined in a 96-hour static test according to the OECD 203 guideline. Under the conditions of the present study, the mean measured 96-h LC50 was determined to be 12 mg/L. The results of the test can be considered reliable without restriction.