[No public or meaningful name is available]

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 July 2008-19 September 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study has been performed according to OECD and/or EC guidelines and according to GLP principles

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] - Part 2: Semi-static method)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] - Part 1: Static Method)

Deviations:

no

Test guideline 5

Qualifier:

according to guideline

Guideline:

other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, December 14, 2000.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

470-280-0

EC Name:

-

Molecular formula:

C26H42O8S

Details on test material:

Name of test material (as cited in study report): SPP100 B7
- Substance type: Highly viscous brownish/yellowish oil
- Physical state: Solid
- Analytical purity: 94.9%
- Lot/batch No.: C0031
- Expiration date of the lot/batch: 31 January 2009
- Stability under storage conditions: Stable
- Storage condition of test material: At room temperature in the dark
- Stability in vehicle:
Water: Not indicated
Acetone: Not indicated
- Solubility in vehicle:
Water: Yes, 0.011 g/l
Aceton: Not indicated

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

During the final test samples for possible analysis were taken from all test concentrations and the control according to the schedule below:
Frequency:
1. At the start of the test and after 72 hours from the freshly prepared solutions.
2. At the first renewal (t=24h) and the end of the test from the 24-hour old solutions. In addition, samples were taken from 100 mg/l on day 2 when all fish had died.
Volume :2 ml from the approximate centre of the test vessels
Storage :samples were stored in a freezer until analysis.

Test solutions

Vehicle:

yes

Details on test solutions:

Preparation of test solutions started with a stock in acetone (SupraSolv., Merck, Darmstadt, Germany) at 1 g/g. Aliquots of 0.2 ml of this stock were added per liter ISO-medium to achieve a loading rate of 100 mg/l. The resulting emulsion was stirred for 2 hours, then stabilized for 30-40 minutes and finally the clear and colourless Water Soluble Fraction (WSF) was siphoned off and used for testing. The lower test concentrations for the final test were prepared by subsequent dilutions of the WSF in test medium.

Test organisms

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp (Cyprinus carpio, Teleostei, Cyprinidae)
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands
- Length at study initiation (length definition, mean, range and SD): Final test: 2.5 ± 0.2 cm
- Weight at study initiation (mean and range, SD): Final test: 0.43 ± 0.11 g
- Feeding during test: no feeding
ACCLIMATION
- Acclimation period: At least 12 days after delivery
- Acclimation conditions (same as test or not): same as the test
- Feeding: Daily with pelleted fish food (Trouvit)
- Health during acclimation (any mortality observed): In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

180 mg CaCO3 per litre

Test temperature:

Between 21.0 and 21.6 ºC

pH:

Between 7.5 and 7.9

Dissolved oxygen:

Between 7.2 and 9.1 mg/l O2

Nominal and measured concentrations:

Nominal concentrations:0, 4.6, 10, 22, 46 and 100% of a WSF prepared at a loading rate of 100 mg/l
Measured concentrations:0 - 46 and 100 % of WSF prep. at 100 mg/l
At the start of the first refreshment period actual test concentrations were 3.4 and 7.2 mg/l at 46 and 100% of the WSF, respectively. At the start of the last refreshment period the actual test concentration was 2.1 at 46% of the WSF. During both 24-hour refreshment periods these concentrations remained stable (i.e. 87-90% of initial). Based on these results, the effect parameters were based on average exposure concentrations, i.e. 2.6 and 6.9 mg/l at 46 and 100% of the WSF.

Details on test conditions:

TEST SYSTEM
- Test vessel: 10 litres, all-glass, containing 9 litres of test solution
- Aeration: The test media were not aerated during the test
- No. of organisms per vessel: 7 fish per concentration and control
- No. of vessels per concentration (replicates):1
- No. of vessels per control (replicates):1
- Biomass loading rate: 0.33 g fish/litre, i.e. 7 fish per 9 litres of test medium

TEST MEDIUM / WATER PARAMETERS
Adjusted ISO medium, formulated using RO-water (tap-water purified by reverse osmosis; GEON Waterbehandeling, Berkel-Enschot, The Netherlands) with the following composition:
CaCl2.2H2O 211.5 mg/l
MgSO4.7H2O 88.8 mg/l
NaHCO3 46.7 mg/l
KCl 4.2 mg/l
- Intervals of water quality measurement: Dissolved oxygen content pH and temperature, Daily in all vessels with surviving fish, beginning at the start of the test (day 0).
OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours photoperiod daily
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :LC50 24,96h and NOEC
TEST CONCENTRATIONS
- Final study: 0, 4.6, 10, 22, 46 and 100% of a WSF prepared at a loading rate of 100 mg/l

Reference substance (positive control):

yes

Remarks:

pentachlorophenol (PCP)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

2.6 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 6.9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

4.2 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI: 2.6 - 6.9 mg/l

Details on results:

Behavioral abnormalities: No abnormalities were observed in the control, 4.6, 10, 22 and 46 % of a WSF prep. at 100 mg/l. Loss of equilibrium , discoloration , hyperactivity were observed in the highest concentration.
- Mortality of control: No mortality in control
- Abnormal responses: no abnormalities were observed
- Effect concentrations exceeding solubility of substance in test medium: no

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- LC50:Under the conditions of the present test PCP induced no lethal effects in carp at 0.10 mg/l. The 96h-LC50 for carp exposed to PCP was 0.20 mg/l (95 % confidence interval between 0.16 and 0.32 mg/l).

Reported statistics and error estimates:

The LC50 could not be determined using the maximum likelihood estimation method with the probits of the percentages of dead fish as function of the logarithms of the corresponding concentrations (Finney, D.J., 1971: Probit analysis, Cambridge University Press, Cambridge, U.K., 3rd edition). This was because there was no concentration between the highest concentration (A) at which 0% mortality and the lowest concentration (B) at which 100% mortality occurred. Instead, the LC50 was calculated as (AB)½, with A and B being limits of the 95% confidence interval.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the present test SPP100 B7 induced no visible effects in carp at or below 2.6 mg/l (NOEC).

The 96h-LC50 was 4.2 mg/l (95% confidence interval between 2.6 and 6.9 mg/l) and was already reached after 48 hours of exposure.