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
Study period:
From 2000-04-12 to 2000-06-08
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP study conducted in compliance with agreed test protocols following OECD Method 202. Acceptable restriction was that analytical measurements on the test material were not conducted.
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:
2000
Report Date:
2000

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Principles of method if other than guideline:
not applicable.
GLP compliance:
yes (incl. certificate)
Remarks:
2000-04-26

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Substance type: amber viscous liquid
- Physical state: liquid
- Analytical purity: no data
- Impurities (identity and concentrations): no data
- Composition of test material, percentage of components: no data
- Isomers composition: no data
- Purity test date: no data
- Lot/batch No.: no data
- Expiration date of the lot/batch: no data
- Stability under test conditions: no data
- Storage condition of test material: at room temperature in the dark
- Other: none

Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
none

Sampling and analysis

Analytical monitoring:
no
Details on sampling:
- Concentrations: 10, 18, 32, 56, 100, 180, 320, 560, 1000 mg/L
- Sample storage conditions before analysis: not applicable
- Time of sampling (stability): 0 and 48 hours.

Test solutions

Vehicle:
no
Details on test solutions:
Amounts of test material (36, 112, 360 and 1120 mg) were each separately dispersed into the vortex of 2 litres of reconstituted water to give the 18, 56, 180 and 560 mg/l loading rates. Further amounts of test material (1 00, 320, 1000, 3200 and 10,000 mg) were each separately dispersed into the vortex of 10 litres of reconstituted water to give the 10, 32, 100, 320 and 1000 mg/l loading rates. These were stirred for 23 hours. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1 hour. As a proportion of the test material was observed to be distributed throughout the water column after standing for 1 hour, the mixtures were filtered through 0.2 pm filters to remove undissolved test material from the aqueous phase to give the 10, 18,32,56, 100, 180,320, 560 and 1000 mg/l loading rate filtered WAFs.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: daphnids
- Source: in-house laboratory cultures.
- Age at study initiation (mean and range, SD): adult
- Weight at study initiation (mean and range, SD): not applicable
- Length at study initiation (length definition, mean, range and SD): not applicable
- Valve height at study initiation, for shell deposition study (mean and range, SD): not applicable
- Feeding during test
- Food type: Chlorella sp.
- Amount: no data
- Frequency: daily, food withheld during exposure
Gravid adults were isolated 24 hours prior to initiation of the test, the yung daphnids produced overnight were removed for testing.

Any immobilisation or adverse reactions to exposure were recorded at 24 and 48 hours after the start of exposure. The criterion of effect used was that Daphnia were considered to be immobilised if they were unable to swim for approximately 15 seconds after gentle agitation.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
none

Test conditions

Hardness:
no data
Test temperature:
20.5°C
pH:
7.8-7.9
Dissolved oxygen:
8.2-8.5 mg O2/L
Salinity:
not applicable
Nominal and measured concentrations:
nominal loading rate: 0, 10, 18, 32, 56, 100, 320, 560, 1000 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: glass jars
- Type (delete if not applicable): static
- Material, size, headspace, fill volume: 250 mL glass jars containing 250 mL test media and covered to reduce evaporation.
- Aeration: no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light:8 h dark
- Light intensity: no data
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
150 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 130-160
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Observations on the test media were carried out during the mixing and testing of the WAFs
At the start of the mixing period, the test material was observed to form cream coloured dispersions of increasing cloudiness with increasing loading rate, each with amber globules of test material circulating around the vortex and throughout the water column.
At the end of the mixing period the loading rates 10 to 180 mg/L were observed to be cream coloured dispersions of increasing cloudiness with increasing loading rate. the 320 to 1000 mg/L loading rates were observed o be cream coloured dispersions with fine amber globules of test material circulating around the vortex and throughout the water column.
At the end of the settling period the 10 to 180 mg/L loading rates were observed to be the same as at the end of stirring whilst the 320 to 1000 mg/L loading rates were cream coloured dispersions with amber globules of test material at the air/water interface and at the sides of the vessels.
After filtration al filtered WAFs were observed to be clear colourless solutions.

Analysis of the test solutions at 0 hours showed measured concentrations (based on total peak area) to range from 0.284 to 6.82 mg/L. At 48 hours the measured concentrations ranged from 0.178 to 6.28 mg/L. These results were in line with the recovery and staability analysis performed on pre-study WAFs. At both the 0 and 48 hurs increasing measured concentrations were shown with increasing loading rates. Inspection of these results indicated a small decline in measured test concentrations over 48 hours for the 10, 32, 320 and 1000 mg/L loading rates. However, given the very low concentrations determined, it was considered that these slightly lower values are due to analytical variation as a result of measuring multiple peaks.
Results with reference substance (positive control):
not applicable
Reported statistics and error estimates:
none

Any other information on results incl. tables

Table 1: Cumulative immobilisation data in the definitve study

Nominal loading rate (mg/L)

Cumulative immobilized Daphnia (n = 10 per replicate)

24 h

48 h

R1

R2

Total

%

R1

R2

Total

%

Control

0

0

0

0

0

0

0

0

10

0

0

0

0

0

0

0

0

18

0

0

0

0

0

0

0

0

32

0

0

0

0

0

0

0

0

56

0

0

0

0

0

0

0

0

100

0

0

0

0

0

0

0

0

180

0

0

0

0

9

8

17

85

320

0

0

0

0

10

10

20

100

560

0

0

0

0

10

10

20

100

1000

0

0

0

0

10

10

20

100

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The 48-Hour EL50 for the test material to Daphnia magna based on nominal loading rates was 150 mg/L loading rate filtered WAF with 95% confidence limits of 130 - 160 mg/L loading rate filtered WAF. The No Observed Effect Loading rate was 100 mg/L loading rate filtered WAF.
Executive summary:

Methods: A study was performed to assess the acute toxicity of the test material to Daphnia magna. The method followed that described in the OECD Guidelines for Testing of Chemicals (1984) No 202, "Daphnia sp, Acute Immobilisation Test and Reproduction Test" referenced as Method C.2 of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC).

Procedures: Following a preliminary range-finding study, twenty daphnids (2 replicates of 10 animals) were exposed to filtered Water Accommodated Fractions (WAFS) of the test material at nominal loading rates of 10, 18, 32, 56, 100, 180, 320, 560 and 1000 mg/L for 48 hours under static test conditions. The number of immobilised Daphnia were recorded after 24 and 48 hours.

Results: The 48-Hour EL50 for the test material to Daphnia magna based on nominal loading rates was 150 mg/L loading rate filtered WAF with 95% confidence limits of 130 - 160 mg/L loading rate filtered WAF. The No Observed Effect Loading rate was 100 mg/L loading rate filtered WAF.

Analysis of the test solutions at 0 hours showed the measured concentrations to range from 0.284 to 6.82 mg/L. At 48 hours the measured concentrations were shown to range from 0.178 to 6.28 mg/L. Measured concentrations were based on total peak area. At both 0 and 48 hours increasing measured concentrations were shown with increasing loading rates. Inspection of these results indicated a small decline in measured test concentrations over 48 hours for the 10, 32, 320 and 1000 mg/L loading rates. However, given the very low concentrations determined, it was considered that these slightly lower values were due to analytical variation as a result of measuring multiple peaks.

Given that the toxicity cannot be attributed to any one or more of the components in the WAF, but to the mixture as a whole, the results are based on nominal loading rates only.