Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008-10-15 to 2009-04-01
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)
Deviations:
no
Qualifier:
according to
Guideline:
other: OECD Series on Testing and Assessment No.23 (Aquatic Toxicity Testing of Difficult Substances and Mixtures, sec. 3.1.2)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material: Reaction product of Acid Yellow 42, Acid Black 2 and Basic Violet 1
- Lot/batch No.: 2008041541
- Expiration date of the lot/batch: 2010-01-31
- Stability under test conditions: see expiry date
- Storage condition of test material: store cool and dry
- Analytical purity: 100 % (96.8 %; UVCB substance)

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: 0.74; 0.39; 0.17; 0.09; 0.04 and 0.02 mg/L
- Sampling method: For the analysis of the test item concentration, samples from the test media of the three highest test concentrations and the
control were taken at the start and at the end of the test. All test medium samples were taken from the approximate centre of the
test vessels after mixing of the test media.
- Sample storage conditions before analysis: The samples were freeze-dried (25-fold concentrated) before analysis.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: WAF method (OECD Series on Testing and Assessment No. 23: Aquatic Toxicity Testing of Difficult Substances and Mixtures, section 3.1.2
- Controls: in the control, test water was used without addition of the test item

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Source: National Institute of Public Health, Hungary
- Age at study initiation (mean and range, SD): less than 24 hours old
- Method of breeding: The Daphnia were bred under similar temperature and light conditions as in the test, and in reconstituted water of a similar
quality regarding to pH, components of the main ions and total hardness as the test water used in the test.
The test animals were fed daily with a few mL of centrifuged algae suspension (Pseudokirchneriella subcapitata).


ACCLIMATION
- Acclimation period:
- Acclimation conditions (same as test or not): Brood Daphnids were maintained in ISO medium under test conditions prior to the start of the test.
- Type and amount of food:
- Feeding frequency:
- Health during acclimation (any mortality observed):


QUARANTINE (wild caught)
- Duration:
- Health/mortality:

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Hardness:
The hardness (CaCO3) of water was: 142 mg/L.
Test temperature:
19.7 – 20.0 °C
pH:
7.28 - 7.87
Dissolved oxygen:
The dissolved oxygen concentration at the end of the test was ≥ 6.3 mg/L in control and test vessels.
Nominal and measured concentrations:
The following nominal concentrations were used in the main study: S.S./10; S.S./20; S.S./40; S.S./80; S.S./160; S.S./320.
The corresponding calculated test item concentrations were: 0.74; 0.39; 0.17; 0.09; 0.04 and 0.02 mg/L.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate (K2Cr2O7)

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
0.042 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95 % CL
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.036 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95 % CL
Duration:
48 h
Dose descriptor:
EC100
Effect conc.:
0.09 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
EC0
Effect conc.:
0.02 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
0.02 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Results with reference substance (positive control):
- Results with reference substance:
24h-EC50 value: 1.59 mg /L , 95 % confidence limits: 1.29 – 1.96 mg /L
In the test with the reference item the results met the expected toxicity of the Potassium dichromate (the 24h EC50 value of the potassium
dichromate should be within the range 0.6 mg/L to 2.1 mg/L). The results are based on the nominal concentrations.
Reported statistics and error estimates:
The EC50 were calculated based on statistical determination (Probit Analysis, statistical software program SPSS+).
The EC100, EC0, NOEC were determined directly from the raw data.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
In this 48-hour static acute toxicity test to Daphnia magna with F 46 Black the 48 h EC50 value was 0.036 mg/L and the 48 h NOEC was determined to be 0.02 mg/L, based on calculated concentrations.
Executive summary:

A 48-hour static acute toxicity test with Daphnia magna according to EU method C.2 and OECD guideline 202 revealed a 48 h EC50 value of 0.036 mg/L and a 48 h NOEC of 0.02 mg/L.