Tar, coal, high-temp.

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:

08 Dec. – 20 Feb. 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07 Aug. – 17 Aug. 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Principles of method if other than guideline:

open system

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Samples for TOC-analysis were taken at 0 and 48 h

Vehicle:

no

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. 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 loading rates of coal tar were weighed on microscope slides, that afterwards were placed 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, 25 mg/L, 50 mg/L, 100 mg/L, 250 mg/L, and 500 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 (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia
- Strain: ---
- 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 were fed with algae (Desmodesmus subspicatus.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

none

Hardness:

see data under "salinity"

Test temperature:

20.2 – 20.4 °C (20 °C ± 2 °C)

pH:

7.8 - 8.0 at 0, 24 and 48 h

Dissolved oxygen:

7.8 - 8.2 mg/L at 0, 24 and 48 h

Salinity:

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 25 mg/L 100 mg/L 500 mg/L (loading)
======================================================================
0 2.3 6.8 7.2 12.7 mg/L (TOC of WAF)
48 3.1 6.3 7.1 11.1 mg/L (TOC of WAF)
=======================================================================
Tar-borne TOC values were about 3.0 - 4.5 mg/L (at 25 mg tar/L), 4.0 - 5.0 mg/L (at 100 mg tar/L) and 8.0 - 10.0 mg/L (at 500 mg tar/L).

Range finding:
Time [h] Control 100 mg/L 1000 mg/L (Loading)
==========================================================
0 1.0 4.2 14.4 mg/L (TOC of WAF)
48 1.3 3.7 11.6 mg/L (TOC of WAF)
==========================================================
The TOC was approx. 2.5 - 3.5 mg/L (at 100 mg tar/L) and 10.5 - 13.5 mg/L (at 1000 mg tar/L).

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): 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
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
- 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

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

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 - 2.5
- Range finding study
- Test concentrations: 100 and 1000 mg/L (loading)
- Results used to determine the conditions for the definitive study: immobilisation 50 % at 100 mg/L, immobilisation 100 % at 1000 mg/L.

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EL50

Remarks:

effective loading

Effect conc.:

72 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EL100

Effect conc.:

250 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOELR

Remarks:

No-observable effect loading rate

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOELR

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Other biological observations: number of moulds incipient reduction from 50 to 100 mg/L
- Mortality of control: 1/20
- 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:

Determination of the EL50 (EL= Effective Loading) was accomplished using Probit-analysis (Finney-method, lognormal distribution; confidence limit 95 %, significance level: p <0.05).

Table 1: Immobilisation rate in dependence of time and loading rate, number of carapace moults, and

number of living animals trapped at the surface [number per total animals, %] (Report Tab. 7)

Control

25 mg/L

50 mg/L

100 mg/L

250 mg/L

500 mg/L

number

[%]

number

[%]

number

[%]

number

[%]

number

[%]

number

[%]

Immobilisation 24 h

0/20

0

0/20

0

0/20

0

4/20

20

4/20

20

8/20

40

Immobilisation 48 h

1/20

5

0/20

0

7/20

35

13/20

65

20/20

100

20/20

100

Moults 48 h

20/20

100

19/20

95

16/20

80

17/20

85

14/20

70

9/20

45

Living animals at the surface 48 h

0/20

0

0/20

0

0/20

0

0/20

0

0/20

0

0/20

0

Validity criteria fulfilled:

yes

Remarks:

- Immobilisation in the control: 0 % / 5 % (nominal value ≤10 %) // - Dissolved oxygen in all vessels: >= 7.9 mg/L / >= 7.8 mg/L (nominal value >= 3 mg/L)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Principles of method if other than guideline:

closed system

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Tar, coal, high-temp.

EC Number:

266-024-0

EC Name:

Tar, coal, high-temp.

Cas Number:

65996-89-6

Molecular formula:

not applicable

IUPAC Name:

naphthalene

Details on test material:

- Name of test material (as cited in study report): Coal Tar
- Molecular formula (if other than submission substance): not applicable
- Molecular weight (if other than submission substance): not applicable
- Substance type: organic
- Physical state: liquid, viscous
- Stability under test conditions: no measured data; based on chemical structure assumed to be stable
- Storage condition of test material: room temperature, exclusion of light

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Samples for TOC-analysis were taken from separate vessels without daphnia at 0 h and 48 h in order to assess the stability of exposure concentrations.

Test solutions

Vehicle:

no

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.
The WAFs were prepared in closed glass beakers (decanter flasks of 2.5 L with screw caps).
The headspace: water ratio was low ( ~ 0.1): The water level was close to the glass fold, i.e. the upper edge of the vertical water column.
(note: The relatively high water volume allowed to weigh 1 mg/L without technical problems.)
The beakers were equipped with a glass tube that allowed the separation of the water phase by siphoning.
The test loadings of coal tar were weighed on microscope slides, that afterwards were placed 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 test vessels were closed immediately following transfer of the WAFs.
- Differential loading: control, 1 mg/L, 2.5 mg/L, 5 mg/L, 10 mg/L, 25 mg/L, and 50 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:

TEST ORGANISM- Common name: Daphnia - Strain: --- 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:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

none

Test conditions

Hardness:

see data under "salinity"

Test temperature:

19.8 – 20.5 °C (20 °C ± 2 °C)

pH:

8.1 - 8.3 at 0, 24 and 48 h

Dissolved oxygen:

8.0 - 8.3 mg/L at 0, 24 and 48 h

Salinity:

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:

nominal: 1, 2.5, 5, 10, 25, and 50 mg/L measured as TOC:
Time [h] Control 1 mg/L 5 mg/L 50 mg/L (loading)
=====================================================================
0 0.3 - 0.4 1.2 1.7 mg/L (TOC of WAF)
48 0.4 - 0.3 0.9 1.7 mg/L (TOC of WAF)
=====================================================================
Tar-borne TOC values were about 0 mg/L (at 1 mg tar/L), 0.5 - 1 mg/L (at 5 mg tar/L) and 1.5 mg/L (at 50 mg tar/L).
Note: The negative TOC-values at the test loading rate of 1 mg/L are derived from the differentiation measurements of
TC (TC = Total Carbon) and TIC (TIC = Total Inorganic Carbon).

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type: closed (sealed with glass stopper)
- Material, size, headspace, fill volume: glass beaker, 100 mL, test volume: 100 mL (no headspace)
- 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

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

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

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 - 2.5
- Range finding study: based on previous daphnia study (open system)

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EL50

Remarks:

(EL = effective loading)

Effect conc.:

2.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EL100

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

NOELR

Remarks:

(No-observable effect loading rate)

Effect conc.:

< 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

48 h

Dose descriptor:

LOELR

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Other biological observations: number of moulds incipient reduction from >5 mg/L
- Immobilisation of control: 1/20
- 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:

Determination of the EL50 (EL= Effective Loading) was accomplished using Probit-analysis (Finney-method, lognormal distribution; confidence limit 95 %, significance level: p <0.05).

Any other information on results incl. tables

Immobilisation rate in dependence of time and loading rate, number of carapace moults, and

number of living animals trapped at the surface [number per total animals, %] (Report Tab. 3)

Control

1 mg/L

2.5 mg/L

5 mg/L

10 mg/L

25 mg/L

50 mg/L

Number

[%]

Number

[%]

Number

[%]

Number

[%]

Number

[%]

Number

[%]

Number

[%]

Immobilisation 24 h

0/20

0

0/20

0

0/20

0

0/20

0

0/20

0

2/20

10

10/20

50

Immobilisation 48 h

1/20

5

6/20

30

10/20

50

13/20

65

13/20

65

18/20

90

20/20

100

Moults 48 h

18/20

90

17/20

85

12/20

60

16/20

80

12/20

60

12/20

60

8/20

40

Living animals at the surface 48 h

0/20

0

0/20

0

0/20

0

0/20

0

0/20

0

0/20

0

0/20

0

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

- Immobilisation in the control: 5 % (nominal value ≤ 10 %) / - Dissolved oxygen in all vessels: >= 8.0 mg/L (nominal value >=3 mg/L)