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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1999-12-20 to 1999-01-04
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP, OECD guideline study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1999
Report date:
1999

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
adopted 1984
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
-
EC Number:
429-290-0
EC Name:
-
Cas Number:
3380-30-1
Molecular formula:
C12 H8 Cl2 O2
IUPAC Name:
5-chloro-2-(4-chlorophenoxy)phenol
Details on test material:
- Name of test material (as cited in study report): FAT 80'220/A
- Physical state: solid powder
- Batch no.: GRU 98
- Expiration date: October 31, 2008
- Purity: >99%
- Solubility in water: 19.5 mg/L (determined in RCC Project 712012)
- Stability in water: >1 year (estimated half-life at 25 °C; determined in RCC Project 712260)
- pH in aqueous solution: 5-7 at a concentration of 0.01 g/L
- Storage conditions: at room temperature at about 20 °C, away from direct sunlight

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
For analytical measurements of the test item concentrations, one sample from the freshly prepared stock solution and duplicate samples from the freshly prepared test media of all test concentrations and the control were taken just before the start of the test (without daphnids) for analysis.
For determination of the stability of the test item under the test conditions, respectively the maintenance of the test item concentrations during the test period, sufficient volumes of the freshly prepared test media of all test concentrations and the control were incubated during the test period under the same conditions as in the actual test (but without daphnids). Duplicate samples were taken at the end of the test period. The collecting of samples after 48 hours from the actual test itself was not possible, since the test media volumes in the test were too small for the analytical requirements. All samples were deep-frozen (at about -20 °C) immediately after sampling.

The concentrations of the test item DCPP were analyzed in the stock solution sample and in the duplicate test media samples from the test concentrations of nominal 0.22 to 0.46 mg/L and both sampling times (0 and 48 hours). The lowest test item concentration of nominal 0.1 mg/L was not analysed, since it was below the 48-hour NOEC. The highest test item concentrations of nominal 1.0 and 2.2 mg/L were not analysed, since after 48-hours the same toxic effect was determined at the next lower test item concentration of nominal 0.46 mg/L. The test item concentration of nominal 1.0 and 2.2 mg/L therefore were considered as being of no biological relevance for the concentration-effect relationship. From the control samples only one of the duplicate samples was analysed from each of both sampling times.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
A stock solution of nominal 10 mg/L was prepared by suspending 10 mg of the test item in about 100 mL test water by ultrasonic treatment for 5 minutes first. The suspension was made up to a volume of 1000 mL with test water and dissolved by ultrasonic treatment for 15 minutes and intense stirring for 3 hours. Adequate volumes of the intensively mixed stock solution were added to test water to prepare the following nominal test concentrations: 0.1, 0.22, 0.46, 1.0, and 2.2 mg/L.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna Straus
- Original source: University of Sheffield/UK in 1992. Since this date the clone is bred in the laboratories of RCC.
- Age at study initiation (mean and range, SD): 6 to 24 hours
- Method of breeding: bred in the laboratories of RCC in reconstituted water of identical quality (regarding pH, main ions and total hardness) and under identical temperature and light conditions as in the tests
- Feeding during test: none



Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Hardness:
2.5 mmol/l (= 250.0 mg/L) as CaCO3
Test temperature:
20-21 °C during the test period
pH:
7.8 to 7.9
Dissolved oxygen:
8.5 mg/L
Nominal and measured concentrations:
- nominal concetration: 0.1, 0.22, 0.46, 1.0 and 2.2 mg/L
- measured concentration: 81 to 87% of the nominal values, therefore all reported values refer to the nominal concentrations
Details on test conditions:
TEST SYSTEM
- Test vessel: 100-mL glass-beaker
- Type (delete if not applicable): closed
- Aeration: no aeration during the test
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted test water
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4:1 (based on molarity)

OTHER TEST CONDITIONS
- Photoperiod: a 16-hour light to 8-hour darkness photoperiod
- Light intensity: light intensity at light period between 200 and 1200 Lux

TEST CONCENTRATIONS
- The test concentrations were based on the results of a 5 day range-finding test and the results of a pre-experiment to the solubility of the test item (without GLP).

Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.32 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 0.22-0.46
Duration:
48 h
Dose descriptor:
EC0
Effect conc.:
0.22 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
0.22 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC100
Effect conc.:
0.46 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
MEASURED CONCENTRATIONS
The analytical mean test item concentrations in the analysed test media varied in the range from 81 to 87% of the nominal values. In the test media the test item was sufficiently stable during the test period of 48 hours. Therefore, all reported biological results were related to the nominal test item concentrations
IMMOBILITY
In the control and up to and including the test item concentration of nominal 0.46 mg/L no immobility or mortality of the test animals or other signs of intoxication were determined during the test period of 24 hours. After 48 hours of exposure the toxicity of the test item to Daphnia magna had increased.
CONTROL ANIMALS
No abnormalities occurred in the control groups.

Any other information on results incl. tables

Influence of DCPP on the mobility of Daphnia magna:

Nominal Concentrations [mg/L]

Number of Daphnia tested/concentration

Number of Daphnia immobilized after 24 and 48 hours (as %)

24 h

48 h

Control

20

0 (0%)

0 (0%)

0.1

20

0 (0%)

0 (0%)

0.22

20

0 (0%)

0 (0%)

0.46

20

0 (0%)

20 (100%)

1.0

20

20 (100%)

20 (100%)

2.2

20

20 (100%)

20 (100%)

Applicant's summary and conclusion