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:
2010-02-15 to 2010-02-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well documented study performed according OECD 201 and GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Sodium vinyl sulphonate solid (Provichem 2202)
- Analytical purity: 25.09%
- Production date: 04/11/2009
- Lot/batch No.: 200911040014
-Shelf-life: 6 months
-Date of receipt: 03/12/2009

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: control; 9.8; 31.3; 100 mg/L
- Sampling method: HPLC-DAD
- Sample storage conditions before analysis: no data

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 99.4 mg of test item was mixed thoroughly with 994 mL of OECD TG 201 medium (stock solution concentration: 100 mg/L). 100 mL of test item was tranferred into sterile 250 mL Erlenmeyer flasks. The control and treated flasks were inoculated with 116 microL of P. subcapitata from the pre-culture to obtain initial cell concentrations of about 1x10000 cells per mL.
- Differential loading:The sample S1 (9.8 mg/L) was 10 times concentrated to fit into dynamic linear range for quantification. S2 (31.3 mg/L) and S3 (100 mg/L) were directly injected into HPLC
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): no vehicle
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): no vehicle
- Evidence of undissolved material (e.g. precipitate, surface film, etc): after preparation, the test item solution was clear and colorless with no insoluble material present

Test organisms

Test organisms (species):
Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM
- Common name: no data
- Strain: SAG 61
- Source (laboratory, culture collection): primary culture obtained from University of Gottingen, Germany
- Age of inoculum (at test initiation): no data
- Method of cultivation: no data

ACCLIMATION
- Acclimation period: pre-culture of P. subcapitata was done three days before initiation of the study
- Culturing media and conditions (same as test or not): same; 6823-6942 lux light intensity at 22.9 to 23.4 °C for three days
- Any deformed or abnormal cells observed: no

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
72 h
Post exposure observation period:
no data

Test conditions

Hardness:
no data
Test temperature:
22.7 - 23.1°C
pH:
test inititation: 8.10-8.15
test termination: 7.02-7.09
Dissolved oxygen:
no data
Salinity:
no data
Nominal and measured concentrations:
Nominal: control; 0.3; 1.0; 3.1; 9.8; 31.3; and 100 mg/L
Measured: analytical verification was carried out in 9.8; 31.3 and 100 mg/L. The mean recovery values were 97.44% at test initiation and 67.24% at test termination.
Details on test conditions:
TEST SYSTEM
- Test vessel: 250 mL flask Erlenmeyer
- Initial cells density: 10000 cells per mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6


GROWTH MEDIUM
- Standard medium used: yes


TEST MEDIUM / WATER PARAMETERS
- according OECD 201

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: to 8.1
- Photoperiod: no data
- Light intensity and quality: 6742-6899 Lux


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: [counting chamber; electronic particle counter; fluorimeter; spectrophotometer; colorimeter]: visually counted using an Improved Neubaur's Haemocytometer under illumination of the microscope after 24, 48 and 72 h after inoculation


TEST CONCENTRATIONS
- Range finding study: 0.1; 1.0; 10; 50; and 100 mg/L
- Test concentrations: 0.3; 1.0; 3.9; 31.3 and 100 mg/L
- Results used to determine the conditions for the definitive study:
Reference substance (positive control):
yes
Remarks:
potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
10.51 mg/L
Nominal / measured:
meas. (geom. mean)
Basis for effect:
other: yield
Remarks on result:
other: 9.32-11.86
Duration:
72 h
Dose descriptor:
EC20
Effect conc.:
0.76 mg/L
Nominal / measured:
meas. (geom. mean)
Basis for effect:
other: yield
Remarks on result:
other: 0.63-0.93
Duration:
72 h
Dose descriptor:
EC10
Effect conc.:
0.19 mg/L
Nominal / measured:
meas. (geom. mean)
Basis for effect:
other: yield
Remarks on result:
other: 0.15-0.26
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
> 55.95 mg/L
Nominal / measured:
meas. (geom. mean)
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
EC20
Effect conc.:
30.15 mg/L
Nominal / measured:
meas. (geom. mean)
Basis for effect:
growth rate
Remarks on result:
other: 24.55-37.03
Duration:
72 h
Dose descriptor:
EC10
Effect conc.:
5.27 mg/L
Nominal / measured:
meas. (geom. mean)
Basis for effect:
growth rate
Remarks on result:
other: 4.20-6.60
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
0.3 mg/L
Nominal / measured:
nominal
Duration:
72 h
Dose descriptor:
LOEC
Effect conc.:
1 mg/L
Nominal / measured:
nominal
Details on results:
- Exponential growth in the control (for algal test): initial cell count 10000 cells/mL and avarage final cell count at 72h was 183 x 10000 cells/mL.
- Observation of abnormalities (for algal test): no data
- Unusual cell shape: no data
- Colour differences: no data
- Flocculation: no data
- Adherence to test vessels: no data
- Aggregation of algal cells: no data
- Any stimulation of growth found in any treatment: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no
Results with reference substance (positive control):
- Results with reference substance valid? yes
- EC50: 0.35 mg/L (95% confidence limit: 0.34 to 0.36 mg/L)
0.94 mg/L (95% confidence limit: 0.91 to 0.97 mg/L)
- Other:
Reported statistics and error estimates:
ECx determination and 95% confidence limits were determined by probit, logit or log-log model (software TOXTAT 3.5 version). NOEC and LOEC were calculated by Bonferroni 't' test using ANOVA techniques.

Applicant's summary and conclusion

Conclusions:
The results of the study showed that SVS at various concentrations had inhibitory effects on the yield and specific growth rate of the algae P. subcapitata. The study reports 72h- NOEC, LOEC, EC50, EC20, EC10.