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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and appropriate guidlines
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
other: EEC, 1989, Dir. 79/831, Annex V, Part. C: Methods for the determination of ecotoxicity, part C2. Acute toxicity to Daphnia magna (updated version of 11/89)
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Principles of method if other than guideline:
The test item is poorly soluble in water and therefore Water Accomodated Fraction (WAF) of the test substance was tested. A toxicity test was performed using 100 mg/l test substance and a control.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
E-AF098T (Batch No. T# 176-3), a white powder. Stored at room temp, dry, closed container.
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
not relevant

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
At the start of the test, a sample of 1 ml was taken from the Water Accommodated Fraction (WAF) of 100 mg/l of test substance in test medium and these without filtration diluted with 9 ml deionized water in the HPLC sample vails.This procedure was repeated for the samples at the end of the test. The amount of test substance present in the test concentrations was quantified by HPLC. The HPLC analyses were performed according to SOP K 19.

Test solutions

Vehicle:
no
Details on test solutions:
The material is poorly soluble, so the Water Accommodated Fraction (WAF) method was used to prepare the test solution. No solvents were used.

The saturated test solution was prepared as follows:
An accurate amount of 0.10009 g test substance was weighed and transfered into a 1L of Dutch Standard Water (DSW) to make 100 mg/l loading. Then the solution was mechanically stirred for 24 h. The undisolved fraction was separated from thesaturated water layer after sedimentation for about 1 h. 50 ml of the visually clear part of the WAF was transferred into the test vessels.


Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
The test was performed with Daphnia magna Clone 5 (water fleas), which were derived from NOTOX at's Hetrogenbosch according to Standard Operation Procedure E1 (ref. 8.3 in the attached report). The animals used in the test were less than 24h old at the beginning of the test and were obtained from parent animals having an age of 2-4 weeks.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
no abnormalities were observed.

Test conditions

Hardness:
ca 200 mg CaCO3/L
Test temperature:
20.0-20.2°C
pH:
Control: 7.6- 8.2
Treatment (WAF 100 mg/L): 8.0 - 7.2 (0h-48h respectively)
Treatment (WAF 1000 mg/L): 8.0 - 7.3 (0h-48h respectively)
Dissolved oxygen:
Control: 8.75-8.81 mg/L (0h-48h respectively)
Treatment (WAF 100 mg/L): 8.15- 8.50 mg/L (0h-48h respectively)
Salinity:
Not relevant
Nominal and measured concentrations:
Samples were quantified using a calibration curve of external standards. More details on analysis, see attched report, pp. 15-16.
Details on test conditions:
The purpose of the test was to determine the acute toxicity of the Water Accommodated Fraction (WAF) of E-AF098T to Daphnia magna (water flea) under continious exposure during 48 hours in a static test.

20 daphnids devided into 4 groups of 5 animals were used.
The daphnids were randomly placed in the test solutions and the test vessels were positioned in a random manner. During the test vessels were covered with glass plates. No aeration of the test substance took place during the test period. The animals were not fed during the test.

The test medium so called Dutch Standard Water (DSW) contained per liter of de-ionized water: 100 mg of NaHCO3, 20 mg of KHCO3, 200 mg of CaCl2*2H2O and 180 mg of MgSO4*7H2O. The test medium wasaerated before being used in the test. The air was water-saturated and purified by an active coal and cotton filter.

The daphnids were exposed to the chosen concentrations of the test substance and immobility and sublethal effects were recorded at approximately 24 and 48 hours. The daphnids were considered immobile when they were not able to swim for 15 seconds after gentle agitation of the test vessel. In addition to immobility, sub-lethal effects such as floating at the surface were recorded.

Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
dissolved
Basis for effect:
other: mortality, appearance and behaivor
Duration:
24 h
Dose descriptor:
EC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
dissolved
Basis for effect:
other: mortality, appearance and behaivor
Duration:
24 h
Dose descriptor:
NOEC
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
dissolved
Basis for effect:
mortality
Remarks:
mortality, appearance and behaivor
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
dissolved
Basis for effect:
other: mortality, appearance and behaivor
Duration:
48 h
Dose descriptor:
EC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
dissolved
Basis for effect:
other: mortality, appearance and behaivor
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
dissolved
Basis for effect:
other: mortality, appearance and behaivor
Details on results:
% Concentration loss over test of: 67% (concentrations of 0.48 mg/L at the beginning of the test and 0.16 mg/L at the end of the test).
Results with reference substance (positive control):
No reference substance was used.
Reported statistics and error estimates:
See pages 15-17 in the attached report.

Any other information on results incl. tables

No immobilization or sub-lethal effects were observed in the preliminary test. Due to the absence of any lethal effects it was not possible to calculate the EC50 and EC100 values.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The results show that WAF of the test substance had no lethal effect on the daphnids. No other sub-lethal effects were observed in the WAF.
Executive summary:

The acute toxicity of the test item to Daphnia magna was tested under static conditions for 48 hours. The results show that WAF of the test substance had no lethal effect on the daphnids. No other sub-lethal effects were observed in the WAF.