Registration Dossier

Ecotoxicological information

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2009-03-12 to 2009-04-01 (draft report)
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.3 (Algal Inhibition test)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Deviations:
no
Qualifier:
according to
Guideline:
other: OECD Series on Testing and Assessment No. 23 (Aquatic Toxicity Testing of Difficult Substances and Mixtures, section 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:
- Sampling method: : For the analysis of the test item concentration, samples from the test media of the three highest test concentration and the
control were taken at the start and at the end of the test.
Three components were measured during the analytical investigation of the test item F 46 Black. Only the three highest
concentrations (S.S./10; S.S./20; S.S./40) were analysed analytically because these 25-fold concentrated solutions were higher than the LOQ of the components.
The measurements of the three lowest concentrations (S.S./80; S.S./160; S.S./320) was not possible, therefore these
concentrations was extrapolated from the Stock Solution. The concentration of the Stock Solution was measured at the start of the test.
- Sample storage conditions before analysis: The samples were freeze-dried (25-fold concentrated) before HPLC analysis.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: WAF method (Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, section 3.1.2
Media preparation methods, Direct addition. OECD Series on Testing and Assessment No. 23)
- Controls: Algal Mineral Salts Test Medium was inoculated (without test item) and examined in parallel in the study.

Test organisms

Test organisms (species):
Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM
- Source (laboratory, culture collection): The algae were supplied by the Georg-August-Universität Göttingen, Albrecht-von-Haller-Institut für
Pflanzenwissenschaften Experimentelle Phykologie und Sammlung von Algenkulturen (SAG),
Nikolausberger Weg 18, D-37073 Göttingen, Germany

Study design

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

Test conditions

Test temperature:
23.8 – 24.4 °C
pH:
pH 7.60 to 7.67 at test start, pH 7.79 to 8.69 at test end
Nominal and measured concentrations:
nominal: S.S./10; S.S./20; S.S./40; S.S./80; S.S./160; S.S./320
measured. 0.142; 0.093; 0.061; 0.058; 0.029 and 0.014 mg/L
Reference substance (positive control):
yes
Remarks:
Potassium dichromate (K2Cr2O7)

Results and discussion

Effect concentrationsopen allclose all
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
0.09 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: 95 % CL
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
0.13 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: 95 % CL
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
0.09 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
other: yield
Remarks on result:
other: 95 % CL
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
0.06 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
0.06 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
0.06 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
other: yield
Duration:
72 h
Dose descriptor:
LOEC
Effect conc.:
0.08 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
LOEC
Effect conc.:
0.08 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
LOEC
Effect conc.:
0.08 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
other: yield
Reported statistics and error estimates:
The EC50 values were determined by Probit Analysis (statistical software program TOXSTAT)
The NOEC and LOEC values were determined directly from the raw data.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
In this 72-h algal growth inhibition test on Pseudokirchneriella subcapitata with F 46 Black, the 72-h EC50 based on growth rate was 0.13 mg/L. The 72-h NOEC was determined to be 0.06 mg/L.
Executive summary:

In this 72-h algal growth inhibition test on Pseudokirchneriella subcapitata with F 46 Black according to EU method C.3 and OECD guideline 201, the 72-h EC50 based on growth rate was 0.13 mg/L. The 72 -h NOEC was determined to be 0.06 mg/L.