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Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2006-04-24 to 2006-05-26
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study without restrictions. The study was conducted according to the appropriate OECD test guideline, and in compliance with GLP.
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Control media and 100 mg/l WAF

- Sampling method: Concentrations of Total Organic Carbon (TOC) were determined in the fresh and old media at the start and end of the first and last 24 hour exposure period.

- Sample storage conditions before analysis: samples were analysed immediately after collection
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: The test substance was a poorly soluble complex mixture and therefore the test media were water-accommodated fractions (WAF) prepared by stirring for 48 hours with a vortex depth not exceeding 10% of the liquid depth. The media were then allowed to stand for 1 hour before drawing off the WAFs for testing.

WAFs were prepared at a nominal loading rate of 100 mg/L. The WAFs were renewed after each successive 24 hour exposure period.

- Controls: Dilution water

- Evidence of undissolved material (e.g. precipitate, surface film, etc): The test substance was not fully soluble at the tested loading rate. However the method of preparing the WAFs was designed to mimimise exposure to undissolved test material.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM

- Common name: Zebrafish

- Source: - Source: Max-Planck-Institut für Entwicklungsbiologie, Tübingen

- Length at study initiation: 3.0 - 3.5 cm

- Feeding during test: none

ACCLIMATION

- Acclimation period: >12 days

- Acclimation conditions: same as test

- Type and amount of food: not reported

- Feeding frequency: not reported

- Health during acclimation (any mortality observed): mortality < 1 %
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
9.0 ºdH
Test temperature:
23 +/- 2.0ºC
pH:
8.0-8.6
Dissolved oxygen:
70-95% ASV
Salinity:
not applicable
Nominal and measured concentrations:
Nominal loading rates: 0(Control) and 100 mg/L WAF

See Tables 2 and 3 for details of measured concentrations.

The test results are reported with reference to nominal loading rate.
Details on test conditions:
TEST SYSTEM

- Test vessel:

- Type: open aquaria

- Material, size, headspace, fill volume: Glass, 20 litre, 10 litre fill volume

- Aeration: none

- Renewal rate of test solution: daily

- No. of organisms per vessel: 7

- No. of vessels per concentration (replicates): 1

- No. of vessels per control (replicates): 1

- Biomass loading rate:


TEST MEDIUM / WATER PARAMETERS

- Metals: Hg/Cd/Pb/Cu/Ni: < 0.001 mg/L

- Nitrate: 4.4 mg/L

- Nitrite: < 0.005 mg/L

Ammonium: < 0.01 mg/L

- Intervals of water quality measurement: daily

OTHER TEST CONDITIONS

- Adjustment of pH:

- Photoperiod: Gro-Lux fluorescent tubes (14 h light, 10 h dark)

EFFECT PARAMETERS MEASURED: daily observations of mortality and behaviour

TEST CONCENTRATIONS

- Spacing factor for test concentrations: limit test
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
water-accommodated fraction loading rate
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
NOELR
Effect conc.:
>= 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
water-accommodated fraction loading rate
Basis for effect:
mortality (fish)
Details on results:
- Behavioural abnormalities: none

- Mortality of control: 0

- Other adverse effects control: none
Reported statistics and error estimates:
No toxic effects were observed in the test and therefore statistical analysis of the results was not required.
Sublethal observations / clinical signs:

Table 1. Test results - percentage mortality for each observation period

Period

Control

100 mg/L

0-24 h start

0

0

0-24 h end

0

0

24-48 h start

0

0

24-48 h end

0

0

48-72 h start

0

0

48-72 h end

0

0

72-96 h start

0

0

72-96 h end

0

0

Table 2. Results of TOC analysis of test media (mg/L)

Period

Control

100 mg/L

0-24 h start

3.2

6.9

0-24 h end

3.4

5.3

72-96 h start

1.9

7.2

72-96 h end

1.7

6.3

Concentrations of Total Organic Carbon (TOC) were determined in the fresh and old Control media and 100 mg/L WAFs at the start and end of the first and last 24 hour exposure period. The measured TOC concentrations ranged between 1.7 and 3.4 mg/L in the Control media and 5.3 and 7.2 mg/L in the 1000 mg/L WAFs. At observation periods 0 – 24 h and 72 – 96 h TOC-values of the Control were relatively stable (within ± 11 %) of initial values. TOC-values in the 100 mg/L WAFs slightly decreased at observation periods 0 – 24 h (23 %) and 72 – 96 h (13 %).

Table 3. Results of GCMS analysis of test media (mg/l)

Vessel

Period

9-Octa-decenoic acid

9,12-Octa-decadienoic acid

Abietic acid

3-Carene

Terpinole

Total

Control

0-24 h start

0,0064

0,014

0,0060

0,00053

0,00054

0,027

Control

0-24 h end

0,0064

0,014

0,0060

0,00053

0,00054

0,027

100 mg/L

0-24 h start

0,0064

0,014

0,0060

0,00053

0,00054

0,027

100 mg/L

0-24 h end

0,0064

0,014

0,018

0,00053

0,00054

0,039

Control

72-96 h start

0,0064

0,014

0,014

0,00053

0,00054

0,036

Control

72-96 h end

0,0064

0,014

0,021

0,00053

0,00054

0,043

100 mg/L

72-96 h start

0,022

0,014

0,21

0,00053

0,00054

0,25

100 mg/L

72-96 h end

0,0064

0,014

0,15

0,00053

0,00054

0,17

Detection Limit

0,0064

0,014

0,0060

0,00053

0,00054

Remark: Quantitative GCMS-values are based on results and/or limits of detection

Concentrations of GCMS-detected components in the Control were below detection limits at start and end of observation period 0 – 24 h. Traces of Abietic acid could be found at start and end of observation period 72 – 96 h. This was probably due to slight residues of Abietic acid in the gas chromatograph and/or from the glass vessels that were used for preparation of the Water Accommodated Fractions.

Concentrations of GCMS-detected components at test loading rate 100 mg/L were below detection limits at the start and end of observation period 0 – 24 h, with the exception of Abietic acid that could be found in traces at the end of this observation period. Traces of 9-Octadecenoic acid could be found at the start of observation period 72 – 96 h. At the end of this observation period concentration of 9-Octa-decenoic acid had decreased below the detection limit (0.0064 mg/l). The decrease was probably caused by biodegradation of this component. Traces of Abietic acid also could be found at the start and end of observation period 72 – 96 h. Total concentrations of GCMS-detected components increased (44 %) during observation period 0 – 24 h and decreased (32 %) during observation period 72 – 96 h.

GCMS concentrations were lower compared to TOC concentrations. This was to be expected because the GCMS-analysis only quantified selected components of the test substance whereas TOC-analysis obtains a sum parameter for all dissolved organic components.

Validity criteria fulfilled:
yes
Conclusions:
A 96-hour LL50 value of >100 mg/L and NOELR of ≥100 mg/L have been determined for the effects of the substance on mortality of Danio rerio. The fish were exposed to water-accommodated fractions of the substance.

Description of key information

The results for short-term toxicity to fish are summarised in Table 7.1.

Reliable data are available for short-term toxicity to fish. A 96-hour LL50 value of >100 mg/L and NOELR of ≥100 mg/L have been determined for the effects of

the substance on mortality of Danio rerio. The fish were exposed to water-accommodated fractions of the substance. The result is supported by similar results from five other studies.

Calculation of PNECs for the aquatic compartment will be based on data for the blocks of constituents rather than on data for the whole substance.

Key value for chemical safety assessment

Additional information