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Ecotoxicological information

Short-term toxicity to fish

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Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study was selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labelling and/or risk assessment.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report Date:
1996

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Remarks:
Conducted according to guideline in effect at time of study conduct
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Purity: 99.7%

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: Samples were collected from each replicate test vessel after 0 and 96 hours (final samples were collected before 96 hours if complete mortality occurred in a test vessel)
- Sampling method: GC equipped with flame ionization detector
- Sample storage conditions before analysis: Not reported

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION: Two 1000 mg/L stock solutions were formulated by combining test substance and dilution water in a glass jar. The test substance was added below the water surface with a syringe. The jars were filled to capacity to eliminate head space and tightly capped. The stock solutions were placed on an orbital shaker adjusted to approximately 150 rpm and incubated at approximately 30°C for 24 hours. Solutions were then incubated at 22.5±1.5°C for 24 hours without shaking. Dilution water used to formulate test media and for the control was shaken and incubated in the same manner as the stock solutions. Following the 48-hour incubation, approximately 50% of each stock solution was carefully removed from the glass jar, and an effort was made to avoid collecting insoluble test substance. The stock solution was used immediately to formulate test media. The dilution water was carbon-filtered, deionized water. The dilution water was stored in polyethylene tanks, where it was aerated and recirculated through particle filters, activated carbon, and an ultraviolet sterilizer.

Test organisms

Test organisms (species):
Pimephales promelas
Details on test organisms:
TEST ORGANISM
- Age at study initiation: juvenile
- Feeding during test: No

ACCLIMATION PERIOD: 5 days

Study design

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

Test conditions

Hardness:
40-48 mg/L
Test temperature:
Temperature ranged from 21.6-22.3°C
pH:
pH ranged from 7.7-8.1
Dissolved oxygen:
Dissolved oxygen ranged from 8.0-8.9 mg/L
Nominal and measured concentrations:
Nominal concentrations: 0, 11, 18, 31, 50, 84, and 140 mg/L (because stock solution contained a greater concentration of test substance than the mean solubility limit of 116 mg/L; probably as the result of the accidental inclusion of undissolved test material, nominal concentrations may be artificially high)
Measured concentrations (initial): Not Detected, 9.26, 14.1, 22.9, 36.8, 60.7, and 108 mg/L (used for all calculations)
Measured concentrations (final): Not Detected, Not Detected, 5.99, 11.6, 32.3, 56.5, 102 mg/L (samples for nominal 50, 84, and 100 mg/L were collected at 24 hours)
Details on test conditions:
TEST SYSTEM
- Test vessel: 3.8-L test vessels that contained 3 L of test solution; because of volatile nature of test substance, test vessels were sealed and not opened during the exposure unless complete mortality occurred
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS:
- Conductivity: Conductivity ranged from 200-210 µmhos/cm

OTHER TEST CONDITIONS
- Photoperiod: 16 hour light and 8 hour dark, with a 15-minute transition period between dark and light
- Light intensity: 45 foot-candles

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality and sublethal effects at 0, 3, 6, 24, 48, 72, and 96 hours

TEST CONCENTRATIONS
- Range finding study results: An initial range-finding test was conducted under static conditions using a dilution water control (0 mg/L) and one concentration of test substance (130 mg/L). At the end of 96 hours, there was 100% survival at 0 mg/L and 40% survival at 130 mg/L. A second range-finding test was conducted under static conditions. The test consisted of a dilution water control and a solvent control (0 mg/L), and 3 nominal concentrations of the test substance (10, 25, and 50 mg/L) prepared using a stock solution prepared in dimethylformamide and 3 concentrations prepared in water (10, 50, and 150 mg/L). At the end of the 96-hour exposure conducted using the dimethylformamide based stock solution, there was 100% survival at 10 mg/L, 80% survival at 25 mg/L, and 60% survival at 50 mg/L. At the end of the 96-hour exposure conducted using the water based stock solution, there was 100% survival at 10 mg/L, 80% survival at 50 mg/L, and 0% survival at 150 mg/L. A third range-finding test was conducted under static conditions and consisted of a control (0 mg/L) and 3 concentrations of test substance (1, 10, and 110 mg/L). At the end of 96 hours, there was 0% survival at 110 mg/L and 100% survival at all other concentrations.
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
27.2 mg/L
Nominal / measured:
meas. (initial)
Basis for effect:
mortality
Remarks on result:
other: 95% confidence interval of 22.9-36.8 mg/L
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
14.1 mg/L
Nominal / measured:
meas. (initial)
Basis for effect:
mortality
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
27.2 mg/L
Nominal / measured:
meas. (initial)
Basis for effect:
mortality
Remarks on result:
other: 95% confidence limit of 22.9-36.8 mg/L
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
27.2 mg/L
Nominal / measured:
meas. (initial)
Basis for effect:
mortality
Remarks on result:
other: 95% confidence limit of 22.9-36.8 mg/L
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
27.2 mg/L
Nominal / measured:
meas. (initial)
Basis for effect:
mortality
Remarks on result:
other: 95% confidence limit of 22.9-36.8 mg/L
Details on results:
- Behavioural abnormalities: Affected fish (6/6) were lethargic and swimming erratically at 24 and 48 hours after exposure to 22.9 mg/L. Affected fish (6/6) were swimming erratically at 72 hours after exposure to 22.9 mg/L. Affected fish (2/6) were lethargic at 96 hours after exposure to 22.9 mg/L.
Reported statistics and error estimates:
Results of the toxicity test were analyzed using standard statistical techniques. The binomial/nonlinear interpolation method was used to calculate LC50 values using the number of dead test organisms and the initial measured concentrations of the test substance. The NOEC is the highest concentration of test substance that allowed at least 90% survival and did not cause sublethal effects.

Any other information on results incl. tables

Mortality Results

 

Initial Measured Concentration (mg/L)

Number Alive

0 hr

3 hr

6 hr

24 hr

48 hr

72 hr

96 hr

0

7

7

7

7

7

7

7

9.26

7

7

7

7

7

7

7

14.1

7

7

7

7

7

7

7

22.9

7

7

7

6

6

6

6

36.8

7

7

7

0

0

0

0

60.7

7

7

7

0

0

0

0

108

7

7

7

0

0

0

0

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The study and the conclusions which are drawn from it fulfil the quality criteria (validity, reliability, repeatability).

96-h LC50 = 27.2 mg/L
Executive summary:

The test was performed under static conditions with 6 concentrations (0, 11, 18, 31, 50, 84, and 140 mg/L) of test substance and a dilution water control at 22.5±1.5°C. The test substance was formulated into a supersaturated stock solution (1000 mg/L) in dilution water, mixed in a sealed container for 24 hours at 30°C, and allowed to equilibrate to test temperature for 24 hours. Water was carefully removed from the mixing vessel to avoid insoluble material and used immediately to initiate the toxicity test. Because the stock solution contained a greater concentration of test substance than the mean solubility limit of 116 mg/L (probably as the result of the accidental inclusion of undissolved test substance), nominal concentrations may be artificially high. Initial measured concentrations were ND, 9.26, 14.1, 22.9, 36.8, 60.7, and 108 mg/L. Because of the volatile nature of the test substance, the test was performed in sealed 3.8 L test vessels that contained 3 L of test solution. Vessels were not opened during the exposure period unless complete mortality occurred. All animals were in good condition at the beginning of the study and 100% of the control fish survived the toxicity test. Exposure of fathead minnows to the test substance resulted in a 96-hour LC50 of 27.2 mg/L, with a 95% confidence interval of 22.9-36.8 mg/L. The 96-hour NOEC was 14.1 mg/L