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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:
17 MAR 2000 - 24 MAY 2000
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:
EEC, 1992, Annex to Commission Directive 92/69/EEC of 31 July 1992 adapting to technical progress for the seventeenth time Council Directive 67/548/EEC on the approximation of laws, regulations and administrative provisions relating to the classification, packing and labeling of dangerous substances (Official Journal No. L 383 A, December 29, 1992), Part C.2. "Acute toxicity for Daphnia magna"
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
OECD Guideline for Testing of Chemicals, No. 202: "Daphnia sp... Acute Immobilisation Test and reproduction Test ", adopted April 4, 1984
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
no
Remarks:
see further explanations in "Any other information on materials and methods incl. tables"
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test medium (reconstituted water and test material) was freshly prepared. Therefore, the calibrated flask with test material and vehicle, reconstituted water, was treated in an ultrasonic device for 1 hour. Subsequently the preparation was aerated, and stirred with a magnetic stirrer for further 23 hours. After 24 hours the formulation was given through a nutsch filter (pore size > 10 - < 16 µm). The filtrate was used for the study.
- Controls: Reconstituted water only was tested as control group.
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): No remarkable observations were made concerning the appearance of the solution of the test material preparation. All test media were clear preparations and stayed unchanged throughout the study.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: Daphnia magna (Straus)
- Age at study initiation (mean and range, SD): Not older than 24 hours.
- Stage and instar at study initiation: Neonates.
- Method of breeding: The strain with the parent generation was bred and maintained in vessels containing a batch of Daphnia magna of different ages.
- Source: Merck KGaA, Darmstadt Institute of Toxicology
- Feeding during test : No.

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES, INCLUDING CULTURING CONDITIONS:
Preparing a new study, juvenile Daphnia magna were separated and placed in 100 ml of reconstituted water. The water was renewed and the Daphnia magna were fed with a suspension of fresh water algae once a week. Newborn animals were separated and allocated to the different dose groups.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
250 mg/L as CaCO3
Test temperature:
The temperature in the control vessel was 21 °C constantly.
pH:
7.88 - 7.98 at test start
7.76 - 7.99 at test end
Dissolved oxygen:
98.3 - 98.4 % at test start
98.9 - 99.2 % at test end
Nominal and measured concentrations:
nominal: 0 and 0.1 g/L
Details on test conditions:
TEST SYSTEM
- Test vessel: glass vessels
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 25 mL all-glass vessels containing 10 mL reconstituted water with or without test material
- Aeration: no
- No. of organisms per vessel: 5 daphnids were introduced in each test vessel
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituuted water was used as dilution water. The water was composited according to ELENDT M4 (1990)
For the reconstituted water the following chemicals (analytical grade) were dissolved in fully demineralized water:
Macro nutrients (mg/L):
CaCl2 * 2H2O = 293.8
MgS04 * 7H2O = 123.3
NaHCO3 = 64.8
KCl = 5.8
Na2SiO3 * 9H20 10
NaNO3 = 0.27
KH2PO4 = 0.14
K2HPO4 = 0.18
Trace elements, macro nutrients and vitamins are added to freshly prepared ISO-medium to reach the following concentrations:
Trace elements (mg/L):
B = 0.5
Fe = 0.2
Mn = 0.1
Li, Rb and Sr = 0.05
Mo = 0.025
Br = 0.0125
Cu and Zn = 0.0063
Co and J = 0.0025
Se =0.0010
V = 0.0003
Na2EDTA * 2H2O = 2.5
Vitamins (µg/L):
Thiamine = 75.0
B12 = 1.0
Biotin = 0.75
After preparation the reconstituted water was aerated for 24 hours.
The hardness: 250 mg/L expressed as CaCO3 and the pH: 7.9 ± 0.3 after an aeration for 24 hours.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light - 8 hours dark regime controlled by a timer
- Other: The study was located in an air-conditioned room in the Institute of Toxicology.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The mobility was determined by visual control and recorded after 24 and 48 hours.

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
- Results used to determine the conditions for the definitive study: not specified
Reference substance (positive control):
yes
Remarks:
The sensitivity of the test system has to be demonstrated periodically. The positive control was carried out with potassium dichromate (Heusener A., 2000).
Duration:
24 h
Dose descriptor:
EC10
Effect conc.:
> 0.1 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: see "Remarks"
Remarks:
The respective measured concentrations were < 0.01 g/L (LOQ).
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 0.1 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: see "Remarks"
Remarks:
The respective measured concentrations were < 0.01 g/L (LOQ).
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
> 0.1 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: see "Remarks"
Remarks:
The respective measured concentrations were < 0.01 g/L (LOQ).
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 0.1 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: see "Remarks"
Remarks:
The respective measured concentrations were < 0.01 g/L (LOQ).
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
>= 0.1 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: see "Remarks"
Remarks:
The respective measured concentrations were < 0.01 g/L (LOQ).
Details on results:
- Behavioural abnormalities: no
- Mortality of control: no
- Other adverse effects control: no
- Immobilisation of control: no
Reported statistics and error estimates:
As the test was conducted as a limit test, no statistical evaluation was necessary. The results were determined directly from the raw data.
Validity criteria fulfilled:
yes
Conclusions:
The study was conducted under GLP according to OECD Guideline 202 and EU Method C.2 on the registered substance itself. The method is to be considered scientifically reasonable with no deficiencies in documentation or deviations from the guidelines, the validity criteria were met. Hence, the results can be considered as reliable to assess the toxicity of the test substance towards daphnids.
Daphnia magna were exposed to an aqueous solution from a test material concentration of nominal 0.1 g/L (limit-test) over 48 hours in an open system under defined conditions. After an exposure to a saturated aqueous solution of a nominal concentration of 0.1 g/L for 48 hours the test animals were not affected. As the analysis of a saturated aqueous solution revealed that the water solubility of the test material was < 0.01 g/L and the limit of quantification of the analytical method was described to be about 0.01 g/L, the test material concentrations in the aqueous medium at the start and the end of the study could not be quantified.
Based on the obtained biological results the 24h and 48h EC50 as well as the 24h and 48h EC10 for Daphnia magna were determined to be > 0.1 g/L, respectively.
Executive summary:

The objective of this study was to determine the acute toxicity of the test item in a static test to Daphnia magna.

For this purpose, juvenile Daphnia magna were exposed to an aqueous test material solution over 48 hours, under defined conditions. The study comprised of four vessels per concentration containing five Daphnia magna , i.e. 20 Daphnia per concentration (test medium group).

Daphnia magna were exposed to an aqueous solution from a test material concentration of 0.1 g/L (limit-test) in an open system.

After an exposure to a saturated aqueous solution of a nominal concentration of 0.1 g/L for 48 hours the test animals were not affected.

The analysis of a saturated aqueous solution revealed that the water solubility of the test material was < 0.01 g/L.

The limit of quantification of the analytical method was described to be about 0.01 g/L. Therefore the test material concentrations in the aqueous medium at the start and the end of the study could not be quantified.

For the test material the following nominal EC50 values for Daphnia magna were calculated:

EC50 (24h) > 0.1 g/L

EC50 (48h) > 0.1 g/L

EC10 (24h) > 0.1 g/L

EC10 (48h) > 0.1 g/L

NOEC 0.1 g/L

Description of key information

Short-term toxicity to aquatic invertebrates: EC10(48h) and EC50(48h) > 0.1 g/L for Daphnia magna based on immobilisation (static, OECD 202, GLP)

Key value for chemical safety assessment

Additional information

The objective of this study was to determine the acute toxicity of the test item in a static test to Daphnia magna.

For this purpose, juvenile Daphnia magna were exposed to an aqueous test material solution over 48 hours, under defined conditions. The study comprised of four vessels per concentration containing five Daphnia magna , i.e. 20 Daphnia per concentration (test medium group).

Daphnia magna were exposed to an aqueous solution from a test material concentration of 0.1 g/L (limit-test) in an open system.

After an exposure to a saturated aqueous solution of a nominal concentration of 0.1 g/L for 48 hours the test animals were not affected.

The analysis of a saturated aqueous solution revealed that the water solubility of the test material was < 0.01 g/L.

The limit of quantification of the analytical method was described to be about 0.01 g/L. Therefore the test material concentrations in the aqueous medium at the start and the end of the study could not be quantified.

For the test material the following nominal EC50 values for Daphnia magna were calculated:

EC50(24h) > 0.1 g/L

EC50(48h) > 0.1 g/L

EC10(24h) > 0.1 g/L

EC10(48h) > 0.1 g/L

NOEC 0.1 g/L