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Diss Factsheets

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 July 2007 - 14 Aug 2007
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Pitch, coal tar, high-temp., < 1% 4- to 5-membered condensed ring aromatic hydrocarbons
EC Number:
Molecular formula:
not applicaple - UVCB
Pitch, coal tar, high-temp., < 1% 4- to 5-membered condensed ring aromatic hydrocarbons
Test material form:
solid: pellets

Sampling and analysis

Analytical monitoring:
Details on sampling:
Samples for TOC-analysis were taken at 0 h and 48 h

Test solutions

Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water Accommodated Fractions (WAFs) prepared by stirring various amounts of the test item in Daphnia-medium
for 24 h with magnetic stirrers (open conditions). The WAFs were prepared in 2-L glass beakers.
The beakers were equipped with a glass tube that allowed the separation of the water phase by siphoning.
The test material was reduced to small pieces with a pestle and then was directly weighed into the glass beakers.
The beakers were then filled with Daphnia-medium.
Mixing was carried out at a speed that was slow enough not to cause dispersion or emulsification of the undissolved fraction
of the test item. To ensure this, the vortex developed at the surface by stirring was set at ~ 10 % of the water depth.
After stirring for 24 h the WAFs were allowed to stand for 1 h before use to facilitate phase separation.
The extracts gained with this method were clear.

- Pre-conditioning: The test vessels of the Definite Daphnia-Test were rinsed with control media and WAFs for ca. 0.5 h before the definite
start of the tests to facilitate adsorption of the test item to the glass walls of the beakers.
Control media and WAFs were then discarded, and the test vessels were refilled again with control media and WAFs, respectively.

- Differential loading: control, 100 and 1000 mg/L (range finding)
control, 100 mg/L
- Controls: without test material (blank; Daphnia medium)
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): none
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
- Common name: Daphnia
- Strain: Clone 5
- Source: origin German Environmental Ministry
- Age at study initiation (mean and range, SD): =< 24 h
- Method of breeding: The animals are cultivated in a climate-controlled room (light/dark = 14 h/10 h; illumination: Gro-Lux fluorescent tubes).

- Feeding during test
- Food type: The animals are fed with algae (Desmodesmus subspicatus

Study design

Test type:
Water media type:
Limit test:
Total exposure duration:
48 h
Post exposure observation period:

Test conditions

see data under "salinity"
Test temperature:
20.1 – 20.2 °C (20 °C ± 2 °C)
7.8 - 8.1 at 0, 24 and 48 h
Dissolved oxygen:
7.2 - 7.6 mg/L at 0, 24 and 48 h
The total of Ca- and Mg-ions was 2.5 mmol/L, the ratio of Ca/Mg was 4:1, the ratio of Na/K was 10:1. The total alkalinity of the solution was 0.8 mmol/L, the pH-value was 7.8 ± 0.2.
Nominal and measured concentrations:
Main test series:
Time [h] Control 1000 mg/L (loading)
0 0.9 0.5 mg/L (TOC of WAF)
48 0.6 0.6 mg/L (TOC of WAF)

Range finding (pretest):
Time [h] Control 100 mg/L 1000 mg/L (Loading)
0 0.6 0.5 0.7 mg/L (TOC of WAF)
48 0.4 0.5 0.7 mg/L (TOC of WAF)

TOC-values appeared to slightly increase with loading range (low water solubility of the test item).
TOC-values were approximately constant over the 48-h test.
Note: Test material-related values are in the range of background values and hence are not precisely quantifiable by the method applied.
Details on test conditions:
- Test vessel:
- Type: open
- Material, size, headspace, fill volume: glass beaker, 50 mL, test volume: 20 mL
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): not applicable

- Source/preparation of dilution water: no data
- Total organic carbon: no data
- Alkalinity: 0.8 mmol/L
- Ca/Mg ratio: 4:1
- Conductivity: no data
- Culture medium different from test medium: no

- Adjustment of pH: no
- Photoperiod: 16 h light, 8 h dark
- Light intensity: no data, Gro-Lux fluorescent tubes

- Spacing factor for test concentrations: ---
- Range finding study
- Test concentrations: 100 and 1000 mg/L (loading)
- Results used to determine the conditions for the definitive study:
immobilisation 0 % at 100 mg/L
immobilisation 10 % at 1000 mg/L
Reference substance (positive control):

Results and discussion

Effect concentrationsopen allclose all
48 h
Dose descriptor:
Effect conc.:
> 1 000 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Remarks on result:
other: 1/20 immobile
48 h
Dose descriptor:
Effect conc.:
ca. 1 000 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Details on results:
- Other biological observations:
- Mortality/immobilisation of control: 0/10 (pretest) and 0/20 (main test)
- Mortality/immobilisation in pretest: 0/10 at 1000 mg/L
- Mortality/immobilisation in main test: 1/20 at 1000 mg/L after 48 h
- Other adverse effects control: none
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none
- Effect concentrations exceeding solubility of substance in test medium: none
Reported statistics and error estimates:
not applicable

Applicant's summary and conclusion

Validity criteria fulfilled:
- Immobilisation in the control: 0 % / 0 % (nominal value ≤10 %) - Dissolved oxygen in all vessels: >= 7.2 mg/L (nominal value >= 3 mg/L)