7-chloro-5-(2-chlorophenyl)-1,3-dihydro-2-oxo-2H-1,4-benzodiazepin-3-yl acetate

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

7-chloro-5-(2-chlorophenyl)-1,3-dihydro-2-oxo-2H-1,4-benzodiazepin-3-yl acetate

EC Number:

220-653-7

EC Name:

7-chloro-5-(2-chlorophenyl)-1,3-dihydro-2-oxo-2H-1,4-benzodiazepin-3-yl acetate

Cas Number:

2848-96-6

Molecular formula:

C17H12Cl2N2O3

IUPAC Name:

7-chloro-5-(2'-chlorophenyl)-2-oxo-2,3-dihydro-1H-1,4-benzodiazepin-3-yl acetate

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

During the test samples for analysis were taken from the undiluted filtrate and from the blank control.
Sampling: Frequency: at t=0h and t=48h.
Volume: 10 ml from the approximate centre of the test vessel.
Storage: The samples not analysed on the day of sampling were stored in a freezer.
Additionally, reserve samples of 10 ml were taken for possible analysis. lf not used, these samples were stored in a freezer for a maximum of three months after delivery of the draft report, pending a the decision of the sponsor concerning additional analysis.

Test solutions

Vehicle:

yes

Details on test solutions:

The test solutions were prepared using a stock solution with a loading rate of 100 mg/L by adding quantitatively 101.4 mg of test substance to 1000 ml ISO-medium. The dispersion was magnetically stirred for 3 days, after which it was a hazy solution with particulates and a surface film. Filtration using a 5 µm paper filter (Schleicher and Schuell) yielded a clear and colourless solution. The filtrate was subsequently diluted in test medium to reach the lower test concentrations.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Start of each batch: With new-born animals, i.e. less than 3 days old, by placing about 250 of them into 10 litres of medium in an all-glass culture vessel.
Maximum age of the cultures: 4 weeks
Renewal of the cultures: After 7 days of cultivation half of the medium twice a week.
Temperature of medium: 18-22°C
Feeding: Daily, a suspension of fresh water algae.
Medium: M7, as prescribed by Dr. Elendt-Schneider (Eiendt, B.P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus. Protoplasma 154, 25-33).

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

250 mg/L expressed as CaCO3

Test temperature:

18-22°C

pH:

6.0-8.5

Dissolved oxygen:

>7 mg/L at the start of the test, >= 5 mg/L at the end of the test

Nominal and measured concentrations:

limit test: loading rate of 100 mg/L corresponding to 4.5 mg/l measured
range-finding test: 0.1, 1.0 and 10 mg/L

Details on test conditions:

Test vessels: 100 ml, all-glass
Medium: ISO, prepared in milli-RO water
Number of daphnia: Iimit test: 20 per concentration, combined range-finding test: 10 per concentration
Loading: 5 per vessel containing 80 ml medium
Light: 16 hours photoperiod daily (355 Iux)
Feeding: No feeding
Aeration: No aeration of the test solutions.
lntroduction of daphnia: Within 1/2 hours after preparation of the test solutions.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 4.5 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

No effects were observed after 24 hours. WY4088 induced a maximum response of immobility, recorded at the maximum soluble concentration (3.3 mg/L), of 25% after 48 hours of exposure.
Both the 24h-EC50 and the 48h-EC50 were above the maximum soluble concentration in test medium, i.e. >3.3 mg/L, corresponding to an average exposure concentration of >4.5 mg/L.
ln conclusion: due to the limited solubility of the test substance, the EC50 could not be reached.

Results with reference substance (positive control):

The reference test was carried out to check the sensitivity of the test system as used by NOTOX. Daphnia were exposed for a maximum of 48 hours to K2Cr2O7 concentrations of 0.10, 0.18, 0.32, 0.56, 1.0 and 1.8 mg/L and to a blank control. Ten daphnia were exposed per concentration. The 24h-EC50 was 1.2 mg/L with a 95% confidence interval between 1.0 and 1.5 mg/L. The 48h-EC50 was 0.60 mg/L with a 95% confidence interval between 0.52 and 0.76 mg/L.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

ln conclusion: The 48h- EC50 was above the maximum soluble concentration in test medium i.e. >3.3 mg/l, corresponding to an average exposure concentration of > 4.5 mg/l