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Ecotoxicological information

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:
5 MAY 2000 - 28 AUG 2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
The study was performed in compliance with the Good Laboratory Practice (GLP) regulations (revised in 1997, ENV/MC/CHEM(98)17). The method followed that described in the OECD Guidelines for Testing of Chemicals (Adopted: 4 April 1984) No 202 "Daphnia sp., Acute Immobilisation Test and Reproduction Test".
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
no
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.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea, Daphnia magna (Straus)
- Age at study initiation (mean and range, SD): not older than 24 h
- Stage and instar at study initiation: Newborn animals were separated and allocated to the different dose groups.
- Source: in house breeding
- Age of parental stock (mean and range, SD): The strain with the parent generation was bred and maintained in vessels containing a batch of Daphnia magna of different ages.
- Feeding during test : no
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
14 ° dH (about 250 mg/L CaCO3)
Test temperature:
22-23 °C
pH:
7.9 +/- 0.3
Dissolved oxygen:
at least 90 %
Nominal and measured concentrations:
limit test at 0.1 g/L
Details on test conditions:
TEST SYSTEM
- Test vessel: glass vessel
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass, 25 mL
- Volume of solution: 10 mL
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per vehicle control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
Reconstituted water according to ELENDT (1990):
Macro nutrients (mg/L):
CaCI2 x 7 H2O 293.80
MgSO4 x 7 H2O 123.30
NaHCO3 64.80
KCI 5.80
Na2SiO3 x 9H2O 10.00
NaNO3 0.27
KH2PO4 0.14
K2HPO4 0.18

Trace elements (mg/L):
B 0.5000
Fe 0.2000
Mn 0.1000
Li, Rb and Sr 0.0500
Mo 0.0250
Br 0.0125
Cu and Zn 0.0063
Co and I 0.0025
Se 0.0010
V 0.0003

Macro nutrients (mg/L):
Na2EDTA x 2H2O 2.50

Vitamins (µg/L):
Thiamine 75.00
B12 1.00
Biotin 0.75


After preparation, the reconstituded water was aerated for 24 hours.

- Hardness: about 14° dH (about 250 mg/L CaCO3)
- pH: 7.9 ± 0.3, after an aeration for 24 hours.

References:

ELENDT, B.-P. Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus.
Protoplasma 154, 25-33, 1990

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light - 8 hours

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : EC50 (24, 48 h)

VEHICLE CONTROL PERFORMED: yes
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 0.1 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 0.1 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Mortality of control: none
- Immobilisation of control: none
In the control and in an aqueous solution of the test material concentration of nominal 0.1 g/L (limit test), no immobilized test animals were observed during the test period of 48 hours.
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.
Results with reference substance (positive control):
The sensitivity of the test system has to be demonstrated periodically. The positive control was carried out with potassium dichromate.

Table 1: Immobilisation of daphnids at a test item concentration of 0 and 100 mg/L (nominal) after 24 and 48 h

Nominal concentration Immobilized/exposed daphnids
[mg/L] 24 hours 48 hours
0 0/20 0/20
100 0/20 0/20
Validity criteria fulfilled:
yes
Conclusions:
The 48 hour EC50 for the test material to Daphnia magna could not be quantified due to the absence of toxicity at the highest test concentration achievable in water. An aqueous solution of 0.1 g/L revealed no aquatic toxicity in the test system.
Executive summary:

The registered substance was tested for toxicity to invertebrates in an acute immobilisation test according to OECD Guideline 202 following GLP.

Purpose
The purpose of this assay was to identify the aquatic toxicity potential of the test material in Daphnia magna to provide a rational basis for hazard estimation for the test item in aquatic environments.

Study Design
For this purpose, juvenile daphnids were exposed to an aqueous test material solution over 48 hours, under defined conditions. The study comprised of four test vessels per concentration and control group containing five daphnids each, i.e. 20 daphnids per concentration (test medium group) and control group.
Daphnids were exposed to a nominal test material concentration of 100 mg/L (limit test) in an open static system.

Results
The analysis of a saturated aqueous solution revealed that the water solubility was < 0.00004 g/L.
The limit of quantification of the analytical method was described to be about 0.000133 g/L. Therefore the test material concentrations in the aqueous medium at the start and the end of the study could not be quantified.

After an exposure to a saturated aqueous solution of a nominal concentration of 0.1 g/L for 48 hours to Daphnia magna the following results were obtained:

Nominal concentration Immobilized/exposed daphnids
[mg/L] 24 hours 48 hours
0 0/20 0/20
100 0/20 0/20


Daphnia magna exposed to an aqueous solution of the nominal test item concentration of 0.1 g/L were not affected.

For the test material, the following nominal EC50 values for daphnids were determined:

24 h EC50     > 100 mg/L
48 h EC50     > 100 mg/L

Conclusion
The 48 hour EC50 for the test material to Daphnia magna could not be quantified due to the absence of toxicity at the highest test concentration achievable in water. An aqueous solution of 0.1 g/L revealed no aquatic toxicity in the test system.

Description of key information

Key, acute immobilisation, D. magna, limit test, OECD 202, GLP: EC50 > 100 mg/L (nominal), NOEC > 100 mg/L (nominal) after 48 h.

Key value for chemical safety assessment

Additional information

Acute immobilisation, OECD 202

The registered substance was tested for toxicity to invertebrates in an acute immobilisation test according to OECD Guideline 202 following GLP.

Purpose
The purpose of this assay was to identify the aquatic toxicity potential of the test material in Daphnia magna to provide a rational basis for hazard estimation for the test item in aquatic environments.

Study Design
For this purpose, juvenile daphnids were exposed to an aqueous test material solution over 48 hours, under defined conditions. The study comprised of four test vessels per concentration and control group containing five daphnids each, i.e. 20 daphnids per concentration (test medium group) and control group.
Daphnids were exposed to a nominal test material concentration of 100 mg/L (limit test) in an open static system.

Results
The analysis of a saturated aqueous solution revealed that the water solubility was < 0.00004 g/L.
The limit of quantification of the analytical method was described to be about 0.000133 g/L. Therefore the test material concentrations in the aqueous medium at the start and the end of the study could not be quantified.

After an exposure to a saturated aqueous solution of a nominal concentration of 0.1 g/L for 48 hours to Daphnia magna the following results were obtained:

Nominal concentration Immobilized/exposed daphnids
[mg/L] 24 hours 48 hours
0 0/20 0/20
100 0/20 0/20


Daphnia magna exposed to an aqueous solution of the nominal test item concentration of 0.1 g/L were not affected.

For the test material, the following nominal EC50 values for daphnids were determined:

24 h EC50     > 100 mg/L
48 h EC50     > 100 mg/L

Conclusion
The 48 hour EC50 for the test material to Daphnia magna could not be quantified due to the absence of toxicity at the highest test concentration achievable in water. An aqueous solution of 0.1 g/L revealed no aquatic toxicity in the test system.