Registration Dossier

Administrative data

Endpoint:
fish early-life stage toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
02 Oct. - 13 Nov. 1991
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (OECD draft)

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:
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

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): benzo(a)pyrene
- Molecular formula (if other than submission substance): C20H12
- Molecular weight (if other than submission substance): 252
- Substance type: organic
- Physical state: solid
- Source: no data
- Analytical purity: no data
- Typical concentration in coal-tar pitch: 1.0 - 1.3 % (data from NL 2008: Coal-Tar Pitch, Annex-XV dossier )
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES (data from NL 2008: Coal-Tar Pitch, Annex-XV dossier, and WHO 1998: EHC 202, if not otherwise indicated)
- Vapour pressure: 7.3*10^-7 Pa (at 25 °C)
- Water solubility: in freshwater, approx. 1.5 - 3.8 µg/L (20 - 25 °C);
soluble portion of the WSF of coal-tar pitch (loading 100 mg/L): [see 4.8: NOACK 2009_WSF, 100 mg/L nominal]
- Henry's law constant: 0.034 Pa*m3/mol (at 25 °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

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 concentrationsopen allclose all
Duration:
42 d
Dose descriptor:
LC50
Effect conc.:
> 4 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Remarks:
(> max. water solubility)
Basis for effect:
mortality
Remarks:
of neonate fish
Duration:
42 d
Dose descriptor:
NOEC
Effect conc.:
4 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Remarks:
(>max. water solubility)
Basis for effect:
larval development
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