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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data is from peer reviewed journal

Data source

Reference
Reference Type:
publication
Title:
Multi-generational effects of four selected environmental oestrogens on Daphnia magna
Author:
Sarah J. Brennan, Concepta A. Brougham, James J. Roche, Andrew M. Fogarty
Year:
2006
Bibliographic source:
Chemosphere 64 (2006) 49–55

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: refer below principle
Principles of method if other than guideline:
Acute toxicity test were performed on Daphnia magna for 24 and 48 hrs according to ISO standards.
GLP compliance:
not specified

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
- Name of test material (as cited in study report): 4-Nonylphenol
- Molecular formula (if other than submission substance): C15-H24-O
- Molecular weight (if other than submission substance): 220.354
- Smiles notation (if other than submission substance): c1(ccc(O)cc1)CCCCCCCCC
- InChl (if other than submission substance): IGFHQQFPSIBGKE-UHFFFAOYSA-N
- Structural formula attached as image file (if other than submission substance): No data available
- Substance type: Organic
- Physical state: Liquid

Sampling and analysis

Analytical monitoring:
not specified

Test solutions

Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: No data available
- Eluate: No data available
- Differential loading: No data available
- Controls: Yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Ethanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): solvent in the stock solution did not exceed 0.1 ml/l
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No data available

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Strain: No data available
- Source: US EPA (Cincinnati, OH).
- Age at study initiation (mean and range, SD): <24 h old
- Weight at study initiation (mean and range, SD): No data available
- Length at study initiation (length definition, mean, range and SD): No data available
- Valve height at study initiation, for shell deposition study (mean and range, SD): No data available
- Peripheral shell growth removed prior to test initiation: No data available
- Method of breeding: No data available
- Feeding during test: not fed during
the course of the experiment
- Food type: unicellular algae Chlorella vulgaris supplemented with Tetrafin fish food
- Amount: 1.4 X 108 cells of algae and 4 mg/l (dry weight) of the fish food suspension
- Frequency: Twice daily

ACCLIMATION
- Acclimation period: No data available
- Acclimation conditions (same as test or not): No data available
- Type and amount of food: No data available
- Feeding frequency: No data available
- Health during acclimation (any mortality observed): No data available

QUARANTINE (wild caught)
- Duration: No data available
- Health/mortality: No data available

Study design

Test type:
not specified
Water media type:
freshwater
Total exposure duration:
48 h

Test conditions

Hardness:
No data
Test temperature:
20 ± 1°C
pH:
No data
Dissolved oxygen:
No data
Salinity:
No data
Nominal and measured concentrations:
No data
Details on test conditions:
TEST SYSTEM
- Test vessel: 50 ml Beaker
- Type (delete if not applicable): No data
- Material, size, headspace, fill volume: 50 ml beakers containing 40 ml of test chemical
- Aeration: No data
- Type of flow-through (e.g. peristaltic or proportional diluter): No data
- Renewal rate of test solution (frequency/flow rate): No data
- No. of organisms per vessel: five
neonatal Daphnia
- No. of vessels per concentration (replicates): No data
- No. of vessels per control (replicates): No data
- No. of vessels per vehicle control (replicates): No data
- Biomass loading rate: No data

TEST MEDIUM / WATER PARAMETERS
No data

OTHER TEST CONDITIONS
- Adjustment of pH: No data
- Photoperiod: 16:8 h photoperiod
- Light intensity: No data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilisation and moulting frequency

TEST CONCENTRATIONS
No data
Reference substance (positive control):
yes
Remarks:
Potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
0.15 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: immobilisation
Remarks on result:
other: if it did not move after 15 s of gentle agitation
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.13 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: immobilisation
Remarks on result:
other: if it did not move after 15 s of gentle agitation
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
0.09 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: moulting frequency
Remarks on result:
other: counting the number of discarded carapaces in each test vessel
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.14 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: moulting frequency
Remarks on result:
other: counting the number of discarded carapaces in each test vessel
Details on results:
No data
Results with reference substance (positive control):
- Results with reference substance valid?: Yes,
- Mortality: Not applicable
- EC50/LC50: EC50 at 24 h of 1.05mg/l
- Other: No data
Reported statistics and error estimates:
ANOVA, Dunnetts test, Levenberg–Marquardt fit model. t-test method, data analysis was performed using MINITAB release 13.2 (MINITAB Inc., PA, USA).

Any other information on results incl. tables

Effects of the test chemicals on mobility of Daphnia magna following 24 h and 48 h exposure:

Chemical

24 h EC50a(mg/l)

48 h EC50a(mg/l)

4-Nonylphenol

0.15 (0.14–0.16)

0.13 (0.12–0.14)

aValues represent EC50 24 h and 48 h with 95% confidence intervals in

parentheses.

Effects of the test chemicals on moulting frequency of Daphnia magna following 24 h and 48 h exposure :

Chemical

24 h EC50a(mg/l)

48 h EC50a(mg/l)

4-Nonylphenol

0.09 (0.04–0.15)

0.14 (0.13–0.16)

aValues represent EC50 24 h and 48 h with 95% confidence intervals in

parentheses.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
Potassium dichromate was the reference chemical. An EC50 24 h of 1.05 (0.97–1.13) mg/l was obtained for potassium dichromate, which is within range of the 0.6–1.7 mg/l stipulated in the ISO standard to ensure test validity
Conclusions:
The EC50 at 48 hrs based on immobilization was found to be 0.13 mg/ml and EC50 at 48 hrs based on moulting frequency was found to be 0.14 mg/ml.
Executive summary:

Acute toxicity test was performed on the Daphnia magna for 48 hrs using 4-nonylphenol. The acute test procedure was carried out in accordance with the ISO guidelines (ISO, 1996) for the determination of the inhibition of mobility of D. magna.

Potassium dichromate was used as the reference chemical to validate acute test Procedures.

Each dilution of test chemical consisted of three 50 ml beakers each containing 40 ml of test chemical and five neonatal Daphnia (newly released neonates (<24 h old) ).The D. magna were not fed during whole experiment and test vessels were maintained for 48 h at 20 ± 1°C in a 16:8 h photoperiod. And measured EC50 for immobilisation and acute moulting frequency at 24 h and 48 h.

 

From the study it was found that the EC50 at 48 hrs based on immobilization was found to be 0.13 mg/ml and EC50 at 48 hrs based on moulting frequency was found to be 0.14 mg/ml.