Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

Link to relevant study record(s)

Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Not specified
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
(see atatchment below)
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
yes
Remarks:
(see attachment below)
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
yes
Details on sampling:
- Sampling method: samples were taken in duplicate at the start of the test (t=0 h) and at the end of the test (t=48 h). Samples were taken agan from the lowest, midlle and highest concentrations. Aliquots of 10 mL were taken in plastic Corning tubes.
- Sample storage conditions before analysis: One sample was transferred to analytical laboratories immediately after sampling. All other samples were stored in a freezer.
Vehicle:
no
Test organisms (species):
Daphnia magna
Details on test organisms:
- The test organism used was the fresh-water crustacean Daphnia magna, cultured in the laboratory under standard conditions.
- Every week a number of cultures is started with ca. 150 daphnids of the same age (about one day) in ca. 4 litres of dilution water. The cultures are fed daily with ca. 4x10^9 algae cells (Chlorella) and ca. 0.13 grams of yeast per 4 litres. The medium is completely replaced at least once a week; at the same time all young born are removed.
- The cultures are kept at 20 ± 1 °C under a 16 h light and 8 h dark regime with transition periods of ca. 30 minutes.
- After 4 weeks the cultures are discarded.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
1.24 - 1.27 mmol/L
Test temperature:
20 ± 1 °C
pH:
The pH was measured at the beginning and at the end of the test and after 24 hours in the exposure media in which all animals were immobile.
Dissolved oxygen:
The oxygen concentrations was measured at the beginning and at the end of the test and after 24 hours in the exposure media in which all animals were immobile.
Nominal and measured concentrations:
0, 5.6, 10, 19, 32, 56, 100 mg/L (nominal) were determined in a preliminary range-finding test.

<0.05, 5.57, 10.1, 18.6, 29.7, 56.6, 102.8 mg/L (measured, t=0 h)
<0.05, 5.37, 8.37, 17.2, 30.9, 53.1, 90.9 mg/L (measured, t=48 h)
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 150 mL all glass beakers each containing 100 mL of exposure medium.
- Aeration: None
- Renewal rate of test solution (frequency/flow rate): None
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The dilution water used was DSWL-E prepared from ground water.
- The nominal composition of the synthetic medium DSWL-E is: Na+ 1.19 mmol.L^-1, K- 0.20 mmol.L^-1, Ca2 2+ 1.36 mmol.L^-1, Mg 2+ 0.73 mmol.L^-1, Cl- 2.72 mmol.L^-1, SO4 2- 0.73 mmol.L^-1 and HCO3- mmol.L^-1.
- This medium is prepared by the addition of several salts to ground water from a locality near Linschoten (the Netherlands). The ground water contains several other trace elements(<< 1 mg.L^-1). Media prepared from it have proved to be suitable for growing several species of water organisms. The equilibrium pH of the medium, after aeration, should be 8.3-8.5, but is usually slightly less, i.e. 8.0-8.2. The hardness, expressed as CaCO3, is about 210 mg.rt .

OTHER TEST CONDITIONS
- Photoperiod: 16h light/ 8h dark

- The mobile, and if applicable the immobile animals, were counted after 24 hours and at the end of the test according to the definition given in the OCED Guideline no. 202. At the same time the immobile animals were removed and the condition of the mobile animals was compared to that of the control animals. The daphnias exposed to 56 and 100 mg/L were also checked under a microscope.

- The actual concentrations of test material in the exposure media were determined by chemical analysis.
Reference substance (positive control):
no
Key result
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
10 mg/L
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
32 mg/L
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
83 mg/L
Basis for effect:
mobility
Remarks on result:
other: 62-111 mg/L (95% confidence limits)
Key result
Duration:
48 h
Dose descriptor:
EC100
Effect conc.:
> 100 mg/L
Basis for effect:
mobility
Details on results:
There were no immobile animals in the control medium at the end of the test. At 56 and 100 mg/L, immobility and lethality were observed at the ends of the test period. At concentrations of 32 mg/L and lower, there were no effects with respect to mobility or lethality except for two dead animals at 19 mg/L. As 10% immobility is allowed for the control and as the mortality observed at 19 mg/L was not dose-related, these observations were not taken into account in estimating the NOEC for mobility in this study. At the end of the test, at 19 mg/L and higher, the condition of the animals was less than that of the controls. The NOEC values for mobility and conditions are therefore 32 mg/L and 10 mg/L respectively.
Results with reference substance (positive control):
not applicable

Variation of pH value: 6.9 -8.2

Lowest measured oxygen concentration: 8.9 O2/L

Variation of temperature measured in one of the control vessels at the start and end of the test respectively: 20.1 -19.6 °C

Table 1: Results of the acute toxicity with Daphnia magna

Parameter (h = hours) Effect Concentration (mg/L)
24h EC50 mobility > 100
48h EC50 mobility 83 (62-111)
48h EC100 mobility > 100
48h NOEC mobility 32
48h NOEC condition 10
Validity criteria fulfilled:
yes
Remarks:
The test exceeded the full validity criteria as requried in the guideline.
Conclusions:
Under the circumstances of the test, the 48h EC50 of the test material for Daphnis magna is 83 mg/L and the 48h NOEC for mobility is 32 mg/L
Executive summary:

The acute toxicity of the test material to the fresh-water crustacean Daphnia magna was determined as described in OECD Guideline no. 202 and the EU Guideline no. C.2 and in compliance with GLP.

The test was carried out as a static test with 4 x 5 daphnids (fourfold exposure media) for the control medium and each concentration. The exposure duration was 48 hours. The nominal concentrations tested were 5.6, 10, 19, 32, 56 and 100 mg/L. The test material was dissolved directly in the diultion water used for the test. The dilution water was also used as control medium.

All exposure media were completely clear throughout the test.

The actual concentration of the test material in the exposure media was determined by chemical analysis. Therefore, samples for chemical analysis were taken from all test concentrations at the start of the test and at the end of the test. At t=0 h, the average measured concentration of the test material in the exposure media was 99% of the nominal value. After 48 hours, the spent solutions contained 92% of the nominal value. As this is above 80% it is allowed to express the results of the test in nominal concentrations. The results of the test were (95% confidence interval in parentheses):

Parameter       Effect       Concentration in mg/l

(h = hours)

24 h EC50       mobility       > 100

48 h EC50       mobility       83 (62 -111)

48 h EC100     mobility       > 100

48 h NOEC     mobility       32

48 h NOEC     condition     10

Under the circumstances of the test, the 48h EC50 of the test material for Daphnis magna is 83 mg/L and the 48h NOEC for mobility is 32 mg/L

Description of key information

Under the circumstances of the test, the 48h EC50 of the test material for Daphnis magna is 83 mg/L and the 48h NOEC for mobility is 32 mg/L.

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
83 mg/L

Additional information

Hooftman & de Wolf, 2003, as amended by Hooftman & de Wolf, 2004, was selected as the key study for this endpoint and was used for classification and labelling purposes.

The acute toxicity of the test material to the fresh-water crustacean Daphnia magna was determined as described in OECD Guideline no. 202 and the EU Guideline no. C.2 and in compliance with GLP.

The test was carried out as a static test with 4 x 5 daphnids (fourfold exposure media) for the control medium and each concentration. The exposure duration was 48 hours. The nominal concentrations tested were 5.6, 10, 19, 32, 56 and 100 mg/L. The test material was dissolved directly in the diultion water used for the test. The dilution water was also used as control medium.

All exposure media were completely clear throughout the test.

The actual concentration of the test material in the exposure media was determined by chemical analysis. Therefore, samples for chemical analysis were taken from all test concentrations at the start of the test and at the end of the test. At t=0 h, the average measured concentration of the test material in the exposure media was 99% of the nominal value. After 48 hours, the spent solutions contained 92% of the nominal value. As this is above 80% it is allowed to express the results of the test in nominal concentrations. The results of the test were (95% confidence interval in parentheses):

Parameter       Effect       Concentration in mg/l

(h = hours)

24 h EC50       mobility       > 100

48 h EC50       mobility       83 (62 -111)

48 h EC100     mobility       > 100

48 h NOEC     mobility       32

48 h NOEC     condition     10

Under the circumstances of the test, the 48h EC50 of the test material for Daphnis magna is 83 mg/L and the 48h NOEC for mobility is 32 mg/L.