Pitch, petroleum, arom.

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

29 Oct. - 19 Nov. 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study, including comprehensive analytics

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.20 (Daphnia magna Reproduction Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Pitch, coal tar, high-temp.

EC Number:

266-028-2

EC Name:

Pitch, coal tar, high-temp.

IUPAC Name:

266-028-2

Constituent 2

Reference substance name:

65996-93-2

Cas Number:

65996-93-2

IUPAC Name:

65996-93-2

Constituent 3

Reference substance name:

Pitch, coal tar, high-temp

IUPAC Name:

Pitch, coal tar, high-temp

Details on test material:

- Name of test material (as cited in study report): coal tar pitch / pitch (coal tar) / pitch, coal tar, high-temp
- Molecular formula (if other than submission substance): not applicable
- Molecular weight (if other than submission substance): not applicable
- Substance type: organic, carbon: > 90 %
- Physical state: solid
- Density: 1.30 – 1.35 g/m3 (20 °C) (DIN 51 757)
- Stability under test conditions: WAF (water accommodated fraction): limited
- Storage condition of test material: room temperature, exclusion of light

- Analysed constituents:
================
Naphthalene
Acenaphthylene
1-Methylnaphthalene
2-Methylnaphthalene
Acenaphthene
Fluorene
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benz(a)anthracene
Chrysene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Dibenz(a,h)anthracene
Benzo(ghi)perylene
Indeno(1,2,3-cd)pyrene
==================

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: no applicable
- Sampling method: on days 0, 2, 6, 14, 18 (fresh media, 0 h) and on days 2, 4, 8, 16, 20 (old media, 48 h).
- Sample storage conditions before analysis: at 6 °C

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances):
- Method: Column elution system as generator system:
- Column: 90 x 15 mm
- Tubing: Tygon®, ID 0.48 mm, length ca. 50 cm and Tygon®, 2.4 mm, length ca. 100 cm
- Storage vessel: brown glass flask (ca. 2.5 L capacity)
Eluate medium: 2 L of dilution water (composition according to Directive 92/69/EEC Method C.2, Annex 1)
Elution time: 48 h
Test material: 200 mg
Peristaltic pump - closed circuit - flow rate: 2 mL/min
Exclusion of light

- Eluate: from powdered pitch (particle size <125 µm, obtained by sieving through an analytical sieve, mesh width 125 µm)
==> saturated solution at the nominal loading of 100 mg/L (water-soluble fraction, WSF)
- Differential loading: no, one loading (nominal 100 mg/L)
- Controls: dilution water without test material
- 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 magna
- Strain/clone: STRAUS / clone 5
- Source: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), Berlin /
breeding continued inhouse
- Age of parental stock (mean and range, SD): 2 - 24 h, obtained by removing the mother animals twice within 22 h.
- Feeding during test
- Food type: mix of Pseudokirchneriella subcapitata (1.65 - 2.35 mL) and Desmodesmus subspicatus (0.690 - 0.950 mL) suspension
- Amount: corresponding to 0.2 mg C per daphnia and day.
- Frequency: at least 5x/wk


ACCLIMATION
- Acclimation period: = continuous breeding
- Type and amount of food: mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata, Desmodesmus subspicatus
and Chlorella vulgaris, with an algae cell density of > 10^6 cells/mL
- Feeding frequency: at least 5x/wk ad libitum

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Remarks on exposure duration:

Water renewal: every 48 hours

Test conditions

Hardness:

157 - 189 mg/L as CaCO3 (Report, Tab. 16)

Test temperature:

20.4 - 22.2 °C [20 ±2 °C ] (Report, Tab. 17)

pH:

7.33 - 7.95 (Report, Tab. 14)

Dissolved oxygen:

8.2 - 9.1 mg/L (Report, Tab. 15)

Nominal and measured concentrations:

nominal: 100 mg/L
analytical (total of 18 PAH): 0.4 - 1.9 µg/L (average 1.0 µg/L)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type: open
- Material, size, headspace, fill volume: Glass beakers (5 cm ID x 8 cm H), 100 mL capacity, 50 mL test volume
- Aeration: no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10
- Biomass loading rate: 1 parent daphnia per 50 mL


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: according to Directive 92/69/EEC Method C.2, Annex 1


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: max. 20 µE×m-2 ×s-1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Behaviour, adult mortality, dry body weight and length of
the parent animals, reproduction parameters (occurrence of aborted eggs and stillborn juveniles,
time of production of first brood, intrinsic rate of natural increase)


RANGE-FINDING STUDY
- Test concentrations:
- Results used to determine the conditions for the definitive study:

Reference substance (positive control):

yes

Remarks:

potassium dichromate, static acute test (24 h) [DIN 38412 L11]

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(loading)

Basis for effect:

other: mortality and reproduction

Effect concentrations 2

Duration:

21 d

Dose descriptor:

LOELR

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(loading)

Basis for effect:

other: mortality and reproduction

Details on results:

- Mortality and immobilisation of parent animals: no
- No. of offspring produced per day per female:
- Body length and weight of parent animals:
- Type and number of morphological abnormalities: none
- Type and number of behavioural abnormalities:
- Number of males and females (parental)
- Time to first brood release: day 8
- Egg development time
- Brood size
- Time to sexual maturity
- Type and magnitude of biochemical changes
- Other biological observations:
- Effect concentrations exceeding solubility of substance in test medium:

Results with reference substance (positive control):

- Results with reference substance valid? yes

The EC50-value of the reference item potassium dichromate after 24 h is within the prescribed concentration range of
1.0 - 2.5 mg/L of quality criteria according to AQS P 9/2 (05/1996) for daphnids clone 5 cultured in Elendt M4 medium.
The EC50-value of the reference item is also within the recommended range of 0.6 - 2.1 mg/L according to OECD-Guideline 202.

- Relevant effect levels: EC50 = 1.50 mg/L (CI: 1.30 - 1.73 mg/L)

Any other information on results incl. tables

There was no biologically significant effect determined in the saturated solution* of the test item, pitch (coal tar).

Table1: Summary of all test item-related effects based on the saturated solution* of pitch (coal tar)

with the nominal loading of 100 mg/L

Effects		Pitch (coal tar)	Significance
	Control	Saturated Solution* (Nominal Loading 100 mg/L)
Mean Number of Juveniles per Producing Parent (Reproduction Rate ± SD)	115±6	114±8	no
Intrinsic Rate of Natural Increase	0.51	0.51	no
Appearance of First Brood [Mean Day]	8.0	8.0	no
Number of Broods	5	5	no
Stillborn Juveniles and Aborted Eggs	0	11	no
Adult Mortality after 21 days [%]	0	0	no
Parent Animals: Mean Dry Weight [mg]	0.83	0.88	no
Parent Animals: Mean Body Length [mm]	5.13	5.15	no

SD = Standard deviation / no = No biological significance

^*) Saturated solution: The maximum dissolved concentration of the test item that can be achieved under the test conditions.

The water-soluble fraction (WSF) of the fresh test medium at the beginning of each 48h test cycle was estimated to range

from 0.82 - 1.90 µg/L (n=5), the WSF in the spent test medium after 48 h was estimated to range from 0.40 - 1.51 µg/L (n=5).

The overall mean was 1.04 µg/L (n=10).

NOTE: For calculation of the mean value of each time interval, the measured concentrations < LOQ_M were set to ½ LOQ_M (15 ng/L).

6 - 14 out of the 18 components were below this LOQ on occasion of analysis (Report Tab. 10).

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

1) Mortality of the parent animal in the control group did not exceed 20 % at the end of the test. 2) The mean number of live offspring produced per parent animal surviving at the end of the test was 115 in the control group.