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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:
disregarded due to major methodological deficiencies
Study period:
19 - 22 Feb 2002
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Significant methodological deficiencies

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): C4-Amin
- Physical state: liquid
- Analytical purity: 98%
- Storage condition of test material: at room temperature (ca. 20 °C) in darkness away from direct sunlight

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: According to the results of a pretest, C4-Amine is not soluble in the test solution. A single water soluble fraction was made and was diluted to create additional test solutions. No auxiliary solvent or emulsifier was used. C4-Amin (100 mg/L) was weighed into test water and sonicated 15 minutes, followed by 48 hours of intensive stirring in the dark. The solution was then filtered under reduced pressure through a 0.45 µm membrane filter (Schleicher and Schuell Type NC45) and adjusted to pH 7.7 using 1N HCl. This stock was used as such for the highest concentration. The stock was diluted 1:2, 1:4, 1:8, 1:16 and 1:32 by adding one part of stock solution to the appropriate volume (1, 3, 7, 15 or 31 parts) of test water.
- Chemical name of vehicle: No vehicle
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none, test medium was a clear solution throughout experiment

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Strain: D. magna Straus
- Source: Clone propagated from original, provided 1992 by University of Sheffield. Clone 5.
- Age at study initiation (mean and range, SD): 6-24 hours, not first brood progeny
- Method of breeding: Organisms bred under same light, temperature, and water quality conditions as in the test
- Feeding during test: No

ACCLIMATION
- Acclimation period: No

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Hardness:
250 mg/L as CaCO3
Test temperature:
21 °C
pH:
7.7 - 7.9
Dissolved oxygen:
8.1 - 8.5 mg/L
Nominal and measured concentrations:
Nominal loading rate: 100 mg/L (undiluted stock), 50 mg/L, 25 mg/L, 12.5 mg/L, 6.3 mg/L, 3.1 mg/L, negative control (untreated test water)
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers (100 mL) with glass covers, filled with 50 mL test medium.
- Aeration: none
- No. of organisms per vessel: Ten
- No. of vessels per concentration: Two
- No. of vessels per control: Two
- Biomass loading rate: < 1 Daphnia/ 2mL

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water (analytical grade salts dissolved in purified water)
294.0 mg/L CaCl2 x 2∙H2O
123.0 mg/L MgSO4 x 7∙H2O
65.0 mg/L NaHCO3
5.8 mg/L KCl
- Alkalinity: 0.8 mmol/l
- Ca/mg ratio: 4:1
- Aeration: Medium was aerated until oxygen saturation was reached prior to study, no further aeration.
- Culture medium different from test medium: Same medium

OTHER TEST CONDITIONS
- Adjustment of pH: Stock solution adjusted from pH 8.6 to pH 7.7 before dilution.
- Photoperiod: 16:8 light:dark, with 30 minute transition periods.
- Light intensity: 200-1200 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilization at 24 and 48 hours. Those organisms not able to swim within 15 s after gentle agitation of the beaker were considered to be immobile.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2-fold dilution
- Range finding study: yes. Results not provided but used to set loading rate for stock solution in the definitive test.
Reference substance (positive control):
no
Remarks:
potassium dichromate tested at least once per year to demonstrate satisfactory test conditions. Test date and results not provided.

Results and discussion

Details on results:
At 48 hours, 100% immobilization was observed in the undiluted stock (100 mg/L nominal loading rate) and 25% immobilization was observed in the 1:2 dilution (50 mg/L nominal loading rate). No immobility or other signs of intoxication were observed at any other test concentration or in the controls. It is not appropriate to calculate effects concentrations from dilutions of a water-soluble fraction having an unknown actual concentation.
Reported statistics and error estimates:
48-hour EC50 could not be calculated from this data

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
48-hour EC50 could not be calculated from this data
Executive summary:

Toxicity of C4-Amin to D. magna was assessed in a limit test conducted according to OECD 202. The test substance was not fully soluble at the limit of 100 mg/L, so a water soluble fraction (WSF) was made. Test substance (100 mg/L) was sonicated in reconstituted test water for 15 minutes, stirred for 48 hours, filtered to 0.45 µm under reduced pressure, and adjusted for pH. A 1:2 dilution series was tested of the WSF stock. Test substance concentrations were not analytically determined. Immobilization at 48 hours was 100% in the undiluted stock and 25% in the 1:2 dilution. No other dilutions displayed any immobilization.

While a toxic effect was observed in this study, it is not appropriate to calculate effects concentrations using nominal concentrations from a dilution series of a water soluble fraction when actual concentration of the WSF is not known. Further, the test substance should have been fully soluble at the concentration tested. This test is considered unreliable. It is not suitable for Risk Assessment, Classification & Labeling, and PBT Analysis.