Pitch, coal tar, high-temp., < 1% 4- to 5-membered condensed ring aromatic hydrocarbons

Long-term toxicity to fish

Administrative data

Endpoint:

fish early-life stage toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 Oct. - 13 Nov. 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)

Version / remarks:

OECD Draft Guideline Fish, Early Life Stage Toxicity Test (1988)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Benzo[def]chrysene

EC Number:

200-028-5

EC Name:

Benzo[def]chrysene

Cas Number:

50-32-8

Molecular formula:

C20H12

IUPAC Name:

benzo[pqr]tetraphene

Test material form:

solid

Details on test material:

- Substance type: organic
Phys.-chem. properties (data from EU 2008: Coal-Tar Pitch, high temperature - Risk Assessment. European Union Risk Assessment Report, The Netherlands, and WHO 1998: EHC 202)
- Vapour pressure: 7.3E-7 Pa (at 25 °C)
- Water solubility: in freshwater, approx. 1.5 - 3.8 µg/L (20 - 25 °C);
- Henry's law constant: 0.034 Pa*m³/mol (at 20 °C)
- log Pow: = 6.13 - 6.5
- pKa: not applicable
- Stability in water: limited due to photolytic decomposition and evaporation
- Stability in light: instable in the presence of UV
- pH dependance on stability: not applicable

Specific details on test material used for the study:

- Source: no data
- Analytical purity: no data
- Stability in water: limited due to photolytic decomposition and evaporation
- Stability in light: instable in the presence of UV

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

weekly

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Organic stock solution in TBA: 120.7 mg B(a)P/L
- Eluate: no (Note: Only for the determination of the water solubility, the elution column technique was used at 25 °C.)
- Differential loading: no
- Controls: with solvent, 84.2 µL TBA/L dilution water
- Chemical name of vehicle (organic solvent): tert. butyl alcohol (TBA)
- Concentration of vehicle in test medium: 84.2 µL TBA/L dilution water
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: commercial hatchery M.B. Ruijsbrook B.V., Maassluis/NL
- acclimatisation: >= 3 wks, 24 °C
- housing: females individual, separated from males, 7 h light, 17 h dark

METHOD FOR PREPARATION AND COLLECTION OF FERTILIZED EGGS
- Numbers of parental fish (i.e. of females used to provide required number of eggs):
1 egg-carrying female placed into a basin with three males previously added (temp. 26 °C)
- Subsequent handling of eggs: deposition in the test vessels, microscopic observation of development

POST-HATCH FEEDING
- Type/source of feed: Immediately after hatching: rotifers; from day 9 Artemia nauplii (enriched with Selco rotifiers)
- Frequency of feeding: daily

Study design

Test type:

other: intermittent flow-through

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

42 d

Test conditions

Hardness:

204 mg/L (as CaCO3)

Test temperature:

24.4 - 25.1 °C

pH:

7.8 - 8.2

Dissolved oxygen:

>= 7.66 mg/L

Nominal and measured concentrations:

nominal: 10 µg/L
analytical (average): 4.0 +-0.4 µg/L (n = 8) (range of maximum water solubility)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 2.6 L chamber containing 4 cylindrical retention chambers with the fish, each 220 mL volume
- Aeration: no
- Type of flow-through: intermittent
- Renewal rate of test solution (frequency/flow rate): every 60 min, renewal of 1 L/h dilution water plus TS dosed from a stock solution.
Water change: about 38 %/h ( 1L/2.6 L)
- No. of fertilized eggs/embryos per vessel: 20
- No. of vessels per concentration (replicates): 4
(At the start of the test, ca. 60 potentially fertilized eggs were placed in each test compartment (total ~240)
and, after first exposure of 24 h, were reduced to about 80 fertilized eggs per test concentration,
thus to assure early exposure in the young blastula stage.)

- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): 4



TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Groundwater supplemented with minerals
- Total organic carbon: 1.5 - 1.7 mg C/L
- Ca/Mg ratio: approx. 2:1


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light / 8 h dark (yellow light)
- Light intensity: no data


EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Hatching success, mortality, and malformations of eggs and larvae


VEHICLE CONTROL PERFORMED: yes

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

42 d

Dose descriptor:

NOEC

Effect conc.:

4 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

larval development

Remarks on result:

other: measured concentration slightly above the max. water solubility

Effect concentrations 2

Duration:

42 d

Dose descriptor:

LC50

Effect conc.:

> 4 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of neonate fish

Remarks on result:

other:

Remarks:

measured concentration slightly above max. water solubility

Details on results:

- Cumulative mortality at each concentration and for each recommended observation time if possible:
Test group: approx. 2.5 % (after 42 d)
- Mortality in the controls: none (day 1 - day 42)
- Development: no effects with regard to hatching, growth or malformations
length: 1.53 +-0.15 cm (test, n = 77) vs. 1.55 +-0.11 cm (control, n = 82) / 1.54 +- 0.11 cm (solvent control, n = 80)
weight: 26.5 +-7.36 mg (test, n = 77) vs. 25.7 +-5.8 mg (control, n = 82) / 25.6 +- 5.7 mg (solvent control, n = 80)

Reported statistics and error estimates:

Mortality: binominal test at 95- and 98-% significance level
Growth: two-tailed Dunnett-test with 95- and 99-% significance level in both cases as compared to controls

Applicant's summary and conclusion

Validity criteria fulfilled:

yes