Anthracene oil

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 Aug. – 19 Sep. 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

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:

open test conditions

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Anthracene oil

EC Number:

292-602-7

EC Name:

Anthracene oil

Cas Number:

90640-80-5

Molecular formula:

Not applicabel

IUPAC Name:

Anthracene oil

Test material form:

other: depending on composition (UVCB) solid or viscous liquid

Details on test material:

anthracene oil; anthracene oil ≤ 50 ppm BaP, AOL (anthracene oil low)
- substance type: organic

Specific details on test material used for the study:

- Name of test material (as cited in study report): anthracene oil (< 50 ppm BaP)
- Lot/batch No. of test material: ATE-No. 10443 (anthracene oil, composite sample No. 07)
- Stability under test conditions: WAF (water accommodated fraction) limited
- 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 at 0 h and 48 h.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water Accommodated Fractions (WAFs) were 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 loading rates of anthracene oil (AOL) 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, 10 mg/L, 25 mg/L, 50 mg/L, 100 mg/L, 250 mg/L, and 500 mg/L (main test)
- 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:

other: static (test system was open to the atmosphere)

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:

20.0 – 20.2 °C (20 °C ± 2 °C)

pH:

7.9 - 8.1 at 0, 24, and 48 h

Dissolved oxygen:

8.0 - 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:
Nominal test concentrations: control, 10 mg/L, 25 mg/L, 50 mg/L, 100 mg/L, 250 mg/L, and 500 mg/L
Measured test concentrations:
Time [h] Control 10 mg/L 100 mg/L 500 mg/L (loading)
==================================================================
0 1.0 1.6 4.3 11.8 mg/L (TOC of WAF)
48 0.7 1.1 3.6 10.2 mg/L (TOC of WAF)
==================================================================
Oil-related TOC values were about 0.5 mg/L (at 10 mg oil/L), 3.0 - 3.5 mg/L (at 100 mg oil/L) and 9.5 - 11.0 mg/L (at 500 mg oil/L).

Range finding:
Nominal test concentrations: 100 mg/L and 1000 mg/L
Measured test concentrations:
Time [h] Control 100 mg/L 1000 mg/L (Loading)
=======================================================
0 1.7 8.9 19.4 mg/L (TOC of WAF)
48 2.4 6.9 16.0 mg/L (TOC of WAF)
=======================================================
The oil-related TOC was approx. 4.5 - 7.0 mg/L (at 100 mg oil/L) and 13.5 - 18.0 mg/L (at 1000 mg oil/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: ca. 1 - 2 mg/L
- 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: 80 % at 100 mg/L
immobilisation: 100 % at 1000 mg/L

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

137 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: EL = effective loading

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: NOELR = no-observable effect loading rate

Effect concentrations 3

Duration:

48 h

Dose descriptor:

LOELR

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: LOELR = lowest observable effect loading rate

Details on results:

- Other biological observations: number of moults significant reduction at 100 mg/L (nominal test concentration) and above
- Immobilisation of control: 0/20
- Other adverse effects control: none
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: not applicable
- Effect concentrations exceeding solubility of substance in test medium: yes; effect concentrations are nominal values (loadings) with WAFs in the range of 0.5 to 18 mg TOC/L (corresponding nominal test material loadings from 10 to 1000 mg/L)

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. 7)

Control

10 mg/L

25 mg/L

50 mg/L

100 mg/L

250 mg/L

500 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

16/20

80

20/20

100

Immobilisation 48 h

0/20

0

0/20

0

0/20

0

1/20

5

4/20

20

18/20

90

20/20

100

Moults 48 h

17/20

85

19/20

95

16/20

80

17/20

85

10/20

50

0/20

0

0/20

0

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: 0 % / 0 % (limit value ≤ 10 %) - Dissolved oxygen in all vessels: ≥ 7.7 mg/L / ≥ 8.0 mg/L (limit value ≥ 3 mg/L)

Conclusions:

Due to the nature of the test substance (UVCB), the fraction dissolved in water (WAF) is different for different loadings (nominal concentrations). Therefore, aqueous toxicity was determined based on loadings.
In a daphnia acute immobilisation test acording to OECD TG 202 with the test substance anthracene oil (BaP content < 50 ppm, AOL), an EL50 (effective loading) of 135 mg/L was determined. In the study, open test conditions (open test vessels) were used.