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Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2011-04-08 till 2011-05-23
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
(2008)
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
(2004)
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
yes
Details on sampling:
- Test solution of 100 mg/L was measured at 0 and 48 hours; control (0 mg/L) at 48 hours
- Sample storage conditions before analysis: Routinely, the samples were analysed on the day of sampling. If this was exceptionally not possible, the samples were stored in a refrigerator at 4 °C until the analysis was carried out.
Vehicle:
no
Details on test solutions:
A direct weighing was prepared to produce the only test concentration. 105.2 mg of the test item were added to 1 litre of dilution water, treated
with an ultra turrax for 60 sec. at 8000 rpm and then stirred for 24 h on a magnetic stirrer. Finally undissolved particles of the test item were
removed by filtration using an aseptic filter with a pore size of 0.45 + 0.2 μm. The pH was measured to be 7.9.
19 mL of the solution were taken and diluted with 1 mL of dilution water containing 10 daphnids resulting in the final concentration. For each test item concentration and the control 2 replicates were prepared.
Test organisms (species):
Daphnia magna
Details on test organisms:
- Name: Daphnia magna STRAUS, parthenogenetic females
- Source: Strain of Bundesgesundheitsamt Berlin
- Maintenance and Acclimatisation: A population of parthenogenetic females of synchronized age structure has been maintained for more than 15 years in the test facility under constant temperature conditions (20 +/- 1 °C) at a 16 : 8 hour light-dark photoperiod (light intensity: < 20 μE x m-2 x s-1). The culture water (so-called 'M4 medium') was partly renewed once a week. The Daphnia were exclusively fed unicellular green algae (Desmodesmus subspicatus) 'ad libitum'. Mortalities of parent Daphnia during the culture period were recorded daily in a semiquantitative way. The neonates were separated from their parent Daphnia by filtration prior to the acute test.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
14.8 °dH (= 264.2 mg/L CaCO3).
Test temperature:
20.7 - 21.1 °C measured at each test vessel at the beginning and the end of the test
pH:
7.9 -8.1 measured at each test vessel at the beginning and the end of the test
Dissolved oxygen:
8.2 - 8.6 mg /L with 92 - 96 % saturation measured at each test vessel at the beginning and the end of the test
Nominal and measured concentrations:
100 mg/L (nominal) plus control
Details on test conditions:
Exposure conditions
- Test vessels: 50 mL glass beakers covered with watch glasses holding 10 neonates in 20 mL of test medium
- Experimental design: 1 test concentration plus 1 control; 10 neonates per vessel, 2 replicates per concentration/control; no feeding during the exposure period; static system
- Method of initiation: neonates were placed in prepared media
- Photoperiod: 16 h light : 8 h dark
- Temperature of incubation unit: 20.4 to 20.5 °C
- Aeration: none
- Test item concentration: 100 mg/L
- Method of administration: direct weighing
- Medium renewal: none
- Duration of exposure: 48 hours
- Criteria of effects: The criterion of adverse effects used in this study was the item-induced alteration of the normal mobility behaviour and the loss of locomotory actions of the neonates, observed at 24 and 48 hours.
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
EL0
Effect conc.:
>= 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EL100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
In the test, Daphnia were exposed to the test item added to dilution water at a limit Effective Loading of nominally 100 mg/L for a period of 48 hours. At this concentration no toxic effects against Daphnia were observed at the end of the 48 hour study period, thus no statistical analysis was required to determine the EL50. Immobilisation rates were recorded at 24-hour intervals. Additionally any abnormal behaviour or appearance of the Daphnia was reported every 24 hours.

No toxic effects against Daphnia were observed at the limit of water solubility.

The test substance is insoluble or poorly soluble in water. Therefore a suitable selective and sensitive chromatographic method for the determination of the test item in aqueous solutions could not be established.

The results are expressed in terms of Effective Loadings (EL). As the test item is a multi constituent and no information about the correlation between molecular weight and the structural formula of the test item are available, a Water Accommodated Fraction (WAF) was used to test effects at a limit concentration of 100 mg/L, and no specific analysis was performed. With the sponsor’s agreement, the content of the test item during the exposure period was verified by DOC determination.

Validity criteria fulfilled:
yes
Remarks:
immobilisation and other abnormalities in the controls did not exceed 10 % by the end of the test; dissolved oxygen concentration remained above 3 mg/L throughout the exposure period
Conclusions:
Daphnia magna were exposed under static conditions for 48 hours to one concentration (100 mg/L) of the test substance. No effect was observed.
Executive summary:

The acute toxicity of the substance to invertebrates Daphnia magna was determined according to the method C.2 "Acute toxicity for Daphnia" (2008) which is equivalent to OECD Guideline for Testing of Chemicals No. 202 "Daphnia sp., Acute Immobilisation Test" (2004).

In the test, Daphnia were exposed to the test item added to dilution water at a limit Effective Loading of nominally 100 mg/L for a period of 48 hours. At this concentration no toxic effects against Daphnia were observed at the end of the 48 hour study period, thus no statistical analysis was required to determine the EL50. Immobilisation rates were recorded at 24-hour intervals. Additionally any abnormal behaviour or appearance of the Daphnia was reported every 24 hours.

The test was conducted as a static test with the test medium unchanged throughout the duration of the test. During the test a temperature range of 18 - 22 °C was maintained in the test vessels, with a maximum temperature fluctuation of +/- 1 °C in each individual test. The temperature, the pH and the oxygen values were measured at the beginning and end of the test.

The test substance is insoluble or poorly soluble in water. Therefore a suitable selective and sensitive chromatographic method for the determination of the test item in aqueous solutions could not be established.

No toxic effects against Daphnia were observed at the limit of water solubility and the immobilisation rate gave a result of EL0 ≥ 100 mg/L and EL100 >100 mg/L. An EL50 of >100 mg/L was calculated.

The results are expressed in terms of Effective Loadings (EL). As the test item is a multi constituent and no information about the correlation between molecular weight and the structural formula of the test item are available, a Water Accommodated Fraction (WAF) was used to test effects at a limit concentration of 100 mg/L, and no specific analysis was performed. With the sponsor’s agreement, the content of the test item during the exposure period was verified by DOC determination.

Description of key information

The acute toxicity to Daphnia magna was determined according to the method C.2 "Acute toxicity for Daphnia" (2008) which is equivalent to OECD Guideline for Testing of Chemicals No. 202 "Daphnia sp., Acute Immobilisation Test" (2004).

Daphnia were exposed to the test item added to dilution water at a limit Effective Loading of nominally 100 mg/L for a period of 48 hours. The test was conducted as a static test with the test medium unchanged throughout the duration of the test. During the test a temperature range of 18 - 22 °C was maintained in the test vessels, with a maximum temperature fluctuation of +/- 1 °C in each individual test. The temperature, the pH and the oxygen values were measured at the beginning and end of the test.

No toxic effects against Daphnia were observed at the limit of water solubility and the immobilisation rate gave a result of EL0 ≥ 100 mg/L and EL100 >100 mg/L. An EL50 of >100 mg/L was calculated.

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
100 mg/L

Additional information

The key value should read EC50 > 100 mg/L as there was no effect in this limit test.