Registration Dossier

Ecotoxicological information

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
long-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:
Chronic 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 g/mol
- 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
Details on sampling:
No data

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): one neonatal Daphnia
- 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:
semi-static
Water media type:
freshwater
Total exposure duration:
21 d
Post exposure observation period:
No data

Test conditions

Hardness:
No data
Test temperature:
No data
pH:
No data
Dissolved oxygen:
No data
Salinity:
No data
Nominal and measured concentrations:
0,0.02,0.04,0.06,0.08,0.10 mg/l
Details on test conditions:
TEST SYSTEM
- Test vessel: 50ml Beaker
- Material, size, headspace, fill volume: Fill volume: 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): three
times weekly
- No. of organisms per vessel: one
- No. of vessels per concentration (replicates): No data available
- No. of vessels per control (replicates): No data available
- No. of vessels per vehicle control (replicates): No data available
- Biomass loading rate: No data available

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: No data available
- Total organic carbon: No data available
- Particulate matter: No data available
- Metals: No data available
- Pesticides: No data available
- Chlorine: No data available
- Alkalinity: No data available
- Ca/mg ratio: No data available
- Conductivity: No data available
- Culture medium different from test medium: No data available
- Intervals of water quality measurement: No data available

OTHER TEST CONDITIONS
- Adjustment of pH: No data available
- Photoperiod: No data available
- Light intensity: No data available

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Adult
Mortality, Cumulative offspring/female
TEST CONCENTRATIONS
- Spacing factor for test concentrations: No data available
- Justification for using less concentrations than requested by guideline: No data available
- Range finding study : Using the NOEC/LOEC results obtained in the acute
studies, a series of chronic studies were established.
- Test concentrations: 0,0.02,0.04,0.06,0.08,0.10 mg/l
- Results used to determine the conditions for the definitive study: No data available
Reference substance (positive control):
not specified

Results and discussion

Effect concentrationsopen allclose all
Duration:
21 d
Dose descriptor:
other: LC100
Effect conc.:
> 0.08 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: decrease in the number of neonatal daphnids
Remarks on result:
other: For first generation
Duration:
21 d
Dose descriptor:
other: LC100
Effect conc.:
> 0.06 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: inhibition in the number of offspring
Remarks on result:
other: For second generation
Details on results:
- Mortality of parent animals: Mention in table
- No. of offspring produced per day per female: No data
- Body length and weight of parent animals: No data
- Type and number of morphological abnormalities: none of the compounds tested significantly impacted the moulting frequencies of D. magna when exposed for 21 d.
- Type and number of behavioural abnormalities: No data
- Number of males and females (parental): No data
- Time to first brood release or time to hatch: No data
- Egg development time: No data
- Brood size: No data
- Time to sexual maturity: No data
- Type and magnitude of biochemical changes: No data
- Other biological observations: : No data
- Effect concentrations exceeding solubility of substance in test medium: : 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

Data and statistical analysis of first generation and second generation Daphnia magna response to 4-nonylphenol:

Concentration (mg/l)

First generation

Second generation

Adult

mortality

Cumulative

moults/femalea

Cumulative

offspring/femalea

Adult

mortality

Cumulative

moults/femalea

Cumulative

offspring/femalea

0.0

0

10.3 ± 1.1

68 ± 9

0

8.9 ± 0.6

72 ± 9

0.02

10

10.0 ± 0.7

70 ± 8

0

8.9 ± 0.7

60 ± 8

0.04

30

9.9 ± 1.2

75 ± 10

40

9.0 ± 0.9

42 ± 10 *

0.06

40

10.5 ± 1.0

59 ± 7

70

10.3 ± 1.5

35 ± 8*

0.08

80

9.0 ± 1.4

48 ± 2*

100

NR

NR

0.10

100

NR

NR

NR

NR

NR

 

NR indicates no result as 100% mortality was observed.

* Significantly different from control at p 6 0.05 (ANOVA, Dunnetts test).

aResults are summarised as the mean of 10 replicates per treatment (±standard deviation) adjusted for mortality.

Applicant's summary and conclusion

Validity criteria fulfilled:
not specified
Conclusions:
The LC100 value for Daphnia magna for first generation and second generation was found to be LC 100>0.08mg/l and LC 100>0.06mg/l.
Executive summary:

Chronic toxicity test was performed on the Daphnia magna for 21 days using 4-nonylphenol. The chronic test procedure was carried out in accordance with the ISO guidelines (ISO, 2000) for long term toxicity testing.

Each dilution of test chemical consisted of ten 50 ml beakers containing 40 ml of test chemical and one neonatal Daphnia. Semi static study design was used for the test and test chemicals were renewed three times weekly. The concentrations used were 0, 0.02, 0.04, 0.06, 0.08, and 0.10 mg/l for first and second generations.

For 4-NP a significant (p ≤ 0.05) first generation effect on fecundity was only seen following exposure to 0.08 mg/l, however, following second generation exposure significant inhibition of the number of offspring produced was seen following exposure to 0.04 and 0.06 mg/l 4-NP.

Therefore, the LC100 value for Daphnia magna for first generation and second generation was found to beLC 100>0.08mg/l and LC 100>0.06mg/l.