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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:
June from 19th to 21th, 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Remarks:
No chemical analysis of the test concentrations was performed, assuming the test item to be stable in medium over the whole test period.
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
adopted 13 April 2004
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Principles of method if other than guideline:
The study was conducted in accordance with ISO/IEC 17025
GLP compliance:
no
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
2 h before the test, vessels were filled with the formulated media to allow equilibration of pH, oxygen and temperature.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna (Straus, 1820).
- Length at study initiation: < 2 mm
- Feeding : 15 g sera micropan (sera GmbH, D-52525 Heinsberg) homogenized in 1 l deionized water.
- Frequency: once a day, except on weekends.

BREEDING SYSTEM
- Vessels: parental and young Daphnia were held in 250 l glass aquaria.
- Photoperiod: 16 h photoperiod day.
- Temperature: 20 ± 2 °C
- Medium: continuously aerated reconstituted water according to ISO; prepared with distilled water (conductivity < 1.5 µS/cm). The final test medium contained 25 mg/l of each of the following four stock solutions: CaCl2 x 2HO 11.76 g/l; Mg So4 x 7 H2O 4.93 g/l; NaHCO3 2.59 g/l; KCl 0.23 g/l. The sum of Ca and Mg ions is 2.5 mmol/l. The bottom of the aquaria is covered with gravel (≤ 1.5 cm). Every month, 25 % of the total volume of the aquaria is replaced by fresh water.
- Alkalinity: 0.8 mmol/l
- Ca : Mg ratio: 4 : 1
- Na : K ratio: 10 : 1
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
0
Test temperature:
20 ± 0.5 °C, determined in a control flask at the start and at the end of the test
pH:
71 - 7.9, determined in all vessels prior to the addition of Daphnia to the test medium and at the end of the test
Dissolved oxygen:
6.5 - 7.1 mg/l, determined in all vessels prior to the addition of Daphnia to the test medium and at the end of the test
Nominal and measured concentrations:
Nominal concentrations: 100, 10 and 1 mg/l
Details on test conditions:
TEST SYSTEM
- Test vessel: 100 ml flasks, all- glass, with 50 ml of test medium.
- No. of organisms per vessel: 20
- No. of organisms per concentration: 40
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Test medium: reconstituted water (ISO 6341), as for breeding; not aerated.
- Aeration: prior to the test, the test medium was aerated vigorously over a period of 2 hours. Final test concentrations were prepared with this aerated water by diluting stock solutions of the test substance.

OTHER TEST CONDITIONS
- Adjustment of pH: the pH was not adjusted.
- Photoperiod: 16 h photoperiod day, supplied by overhead white fluorescent tubes.
- Temperature: 20 ± 0.5 °C (temperature-controlled water bath).

EFFECT PARAMETERS MEASURED
The median effective concentration is the concentration estimated to immobilize 50 % of the Daphnia after 24 or 48 hours of exposure. Those individuals not able to swim within 15 seconds after gentle agitation of the test vessel were considered to be immobile.
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
After 24 and 48 hours of incubation no significant effects ( < 10 % of a total of 40 individuals) were observed at all test concentrations and in the blank control, respectively. The experimentally determined no-effect concentration (NOEC, EC10) was ≥ 100 mg/l after an exposure of 24 and 48 hour, respectively.

Immobilization of Daphnia magna after 24 h and 48 h of test material exposure

Nominal concentration (mg/l) Code No. Individuals per test vessel Immobile Daphnia (N./vessel) Immobile Daphnia (N./vessel) % Immobile Daphnia after 24 h of exposure % Immobile Daphnia after 48 h of exposure
Control A 20 0 0 0 0
0 B 20 0 1 0 5
C 20 0 0 0 0
1 A 20 0 0 0 0
B 20 1 1 5 5
10 A 20 0 1 0 5
B 20 1 2 5 10
100 A 20 0 1 0 5
B 20 0 1 0 5

pH values and oxygen concentrations at the start (0 h) and end (48 h) of the test

Nominal concentration (mg/l) Code pH values
0 h
pH values
48 h
Oxygen conc. (mg/l) 0 h Oxygen conc. (mg/l) 48 h
Control A 7.1 7.8 7.1 6.5
0 B
C
1 A 7.2 7.9 6.6 6.5
B
10 A 7.2 7.9 6.6 6.5
B
100 A 7.4 7.9 6.7 6.5
B

Values were determined in the combined fractions only.

Validity criteria fulfilled:
yes
Remarks:
in the control not more that 10 % of the daphnids were immobilised and the dissolved oxygen concentration at the end of the test was 3 mg/l in control and test vessels
Conclusions:
EC50 (48h) > 100 mg/l (nominal)
Executive summary:

The acute toxicity of the test compound to Daphnia magna was determined according to the OECD Guideline 202. The test was carried out under static exposure conditions for a period of 48 hours. A screening test with nominal concetrations of 100, 10 and 1 mg/l, was performed.

No analytical verification of the test material was conducted. The evaluation of the test was based on the nominal concentrations of the test material, assuming the test compound to be stable in water over 48 hours.

After 24 and 48 hours of incubation no significant effects ( < 10 % of a total of 40 individuals) were observed at all test concentrations and in the blank control, respectively. The experimentally determined no-effect concentration (NOEC, EC10) was ≥ 100 mg/l after an exposure of 24 and 48 hour, respectively. The nominal EC50 of the test compound to Daphnia magna for 24 hours and 48 hours was stated to be greater than 100 mg/l at both observation periods.

Conclusion

EC50 (48h) > 100 mg/l (nominal)

Description of key information

Non harmful/toxic to invertebrates after short-term exposures (EC50 (48h) > 100 mg/l (nominal))

Key value for chemical safety assessment

Additional information

The acute toxicity of Direct Blue 199 Na to Daphnia magna was determined according to the OECD Guideline 202. The test was carried out under static exposure conditions for a period of 48 hours, at the following concentrations: 1, 10 and 100 mg/l. After 24 and 48 hours of incubation no significant effects (< 10 % of a total of 40 individuals) were observed at all test concentrations and in the blank control, respectively.

No analytical verification of the test material was conducted and the evaluation of the test was based on the nominal concentrations of Direct Blue 199 Na, assuming the test compound to be stable in water over 48 hours.