6-chloro-5-(2-chloroethyl)-1,3-dihydroindol-2-one

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

Study period:

5 March 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: done under GLP and OECD method

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Principles of method if other than guideline:

The study was conducted in accordance with EEC Methods for Determination of Ecotoxicity Annex
to Directive 92/69/EEC (O.J. No. L383A, 29.12.92) Part C, Method 2 "Acute toxicity to Daphnia" and the OECD Guideline for Testing of Chemicals No. 202, Part I "Daphnia Acute Immobilisation Test".
Test was assessed under static exposure conditions.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

6-chloro-5-(2-chloroethyl)-1,3-dihydroindol-2-one

EC Number:

421-320-0

EC Name:

6-chloro-5-(2-chloroethyl)-1,3-dihydroindol-2-one

Cas Number:

118289-55-7

Molecular formula:

Hill formula: C10 H9 Cl2 N O CAS formula: C10 H9 Cl2 N O

IUPAC Name:

6-chloro-5-(2-chloroethyl)-2,3-dihydro-1H-indol-2-one

Test material form:

not specified

Details on test material:

Expiry: 2 June 1999
Purity: 98.82% by hplc
Date received: 2 June 1997

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

TEST SUBSTANCE PREPARATION: concentrations were verified by chemical analysis. Samples, 100 ml, were taken from the pooled replicates of control and each test concentration at 0 hours and after 48 hours

Test solutions

Vehicle:

no

Details on test solutions:

The test organisms were maintained and the tests conducted in a reconstituted medium Elendt M4. The medium was prepared using analytical grade reagents and deionised water. Appendix 2 gives full details of the medium

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Strain:Straus
- Source:cultured in-house and were obtained from a strain originating from the Institute National de Recherche Chimique Applique (IRChA), France
- Age at study initiation (mean and range, SD):less than 24 hours old

ACCLIMATION
- Acclimation period:None

Stock cultures of Daphnia magna were maintained in glass vessels containing approximately 300 ml of Elendt M4 culture medium with a 16 hour light: 8 hour dark photoperiod at 20 ± 2°C. Cultures were fed daily with a suspension of the unicellular green alga, Chiorella vulgaris. Culture conditions ensure that cultures reproduce by parthenogenesis.

The day before the start of the study, all juvenile Daphnia were removed from the laboratory cultures. The following morning, juveniles produced by the gravid (egg-bearing) adult Daphnia were used in the test

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

without renewal of test media

Post exposure observation period:

The temperature in each vessel was measured daily and the pH and dissolved oxygen levels recorded at the start and at the end of the study

Test conditions

Hardness:

No Data

Test temperature:

20 ± 1 °C

pH:

pH 7.8

Dissolved oxygen:

6.8-8.4 mg/l

Nominal and measured concentrations:

The following test concentrations were prepared based on the results of the earlier range finding study.

Mean measured test concentrations: 0.044, 0.098, 0.23, 0.49, 1.1, 2.3 and 5.0 mg/L

Nominal exposure concentrations quoted in this report refer to the test material as received; no allowance has been made for a purity of less than 100%.

Details on test conditions:

TEST SYSTEM
- Test vessel:250 ml capacity glass jars.
- Type (delete if not applicable): loosely covered to minimise evaporative losses
- Material, size, headspace, fill volume:
- Aeration:
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate):
- No. of organisms per vessel:
- No. of vessels per concentration (replicates):
- No. of vessels per control (replicates):
- No. of vessels per vehicle control (replicates):
- Biomass loading rate:Ten first instar Daphnia were placed without conscious bias into glass jars

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod:with a photoperiod of 16 hours light: 8 hours dark and without supplementary aeration or feeding
- Light intensity:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations:
- Results used to determine the conditions for the definitive study:

Reference substance (positive control):

not specified

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 5 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.51 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 0.34-0.75 (mg/l)

Details on results:

Highest test concentration resulting in 0% immobilisation: <0.044 mg/1*
Lowest test concentration resulting in 100% immobilisation:5.0 mg/1

*A 10% immobilisation was observed in the control and lowest test level. However, a 10% immobilisation is not considered to be biologically significant, and the "No observed effect concentration" is assumed to be 0.044 mg/1.

Any other information on results incl. tables

Summary of chemical analysis results

Occasion	Nominal concentration(mgll)	Measured concentration(mg/1)	Achieved dilutionfactor(%)!	%oftarget dilutionfactor
0hours	Control 1.0 2.2 4.6 10 22 46 100	ND 0.04580 0.1057 0.2285 0.4999 1.105 2.293 4.945	- 0.916 2.11 4.57 10.0 22.1 45.9 98.9	- 91.6 96.1 99.3 100 100 99.7 98.9
48 hours	Control 1.0 2.2 4.6 10 22 46 100	ND 0.04264 0.08979 0.2345 0.4809 1.109 2.312 5.007	- 0.853 1.80 4.69 9.62 22.2 46.2 100	- 85.3 81.6 102 96.2 101 101 100

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 48 h EC50 (immobilisation value for CP-89,575 with Daphnia magna) was determined to be 0.51 mg/1. The 'No-observed effect concentration' (NOEC) was 0.044 mg/1.