Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
07 August 1985 to 11 August 1985
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Analytical verification of the test item is not considered to be fully reliable.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1985
Report Date:
1985

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
not applicable
GLP compliance:
yes
Remarks:
EPA Good Laboratory Practice Regulations set forth in 40 CFR Part 792

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch I

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: The stability, identity, strength, and composition or other characteristics which appropriately identify the test substance are the responsibility of the Sponsor.

FORM AS APPLIED IN THE TEST (if different from that of starting material): Water soluble fraction of MRD-84-521

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: The BW2 (The dilution water used during studies in the Environmental Toxicology Laboratory is a blend of reverse osmosis (RO) water and carbon filtered well water. For this study the ratio was approximately 1.8:1 RO to well water (BW2)) used for stock solution preparation and 100% stock solutions were sampled during the study.
- Sampling method: The aqueous samples received from the Environmental Toxicology Department were analyzed for Total Carbon (TC) by direct injection into the Dohrmann DC-80 Total Carbon Analyzer. At least two injections were made of each sample and only the mean value is reported.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The water soluble fraction of MRD-84-521 was prepared by combining the test material and BW2 at a ratio of approximately 1:150. The mixture was then stirred slowly for approximately 72 hours. After the mixing period, the WSF was drawn off by means of a sampling tube. The MRD-84-521 test material water soluble fraction
was administered in aqueous solution via a diluter system. The test chambers were replicate 15 liter tanks containing approximately 14 liters of test solution.

Test organisms

Test organisms (species):
Pimephales promelas
Details on test organisms:
TEST ORGANISM
- Common name: Fathead Minnow
- Strain: Lot 41
- Source: Cultured in the Environmental Laboratory
- Age at study initiation (mean and range, SD): Approximately 25 weeks
- Length at study initiation (length definition, mean, range and SD): Mean length: 2.5 cm, SD 0.3
- Weight at study initiation (mean and range, SD): Mean weight: 0.276 g, SD 0.10

ACCLIMATION
- Acclimation period: at least 14 days prior to study
- Acclimation conditions (same as test or not): similar (Fish are kept in holding tanks with dilution water (BW2) similar in characteristics to that which was used during the test phase; fish were held in flow-through conditions prior to the test.)
- Type and amount of food during acclimation: Fish were fed ad libitum during normal holding, Rangen Salmon Starter and brine shrimp (Artemia salina)
- Health during acclimation (any mortality observed): none

FEEDING DURING TEST
- Food type: Fish were not fed during the study

Study design

Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h

Test conditions

Hardness:
158 mg CaCO3 /L
Test temperature:
Mean: 21.78 +/- 0.26°C
Range: 21.0 - 22.0 °C
pH:
Range: 7.6 - 8.0
Dissolved oxygen:
Range: 7.7 - 8.6 mg/L
Nominal and measured concentrations:
Nominal: Control, 6.25, 12.5, 25.0, 50.0, 100.0 % WSF
Measured: The Total Carbon analyses for the stock solution batches used during this study indicated that the test material water soluble fraction could not be measured in solution. The difference in Total Carbon between the BW2 and 100% stock falls into the realm of analytical variation for that method. As a result, the treatments were run nominally and no further concentration analysis was performed.
Details on test conditions:
TEST SYSTEM
- Test vessel: tanks
- Material, size, headspace, fill volume: size: 15 L, fill volume: 14 L
- Renewal rate of test solution (frequency/flow rate): Dwell time: 40 min; total cycle time: 3 min 6 seconds
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2 replicates
- No. of vessels per control (replicates): 2 replicates
- Biomass loading rate: approximately 0.023g fish/liter/24 hours

TEST MEDIUM / WATER PARAMETERS
- Culture medium different from test medium: no
- Intervals of water quality measurement: semi-annually

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: 16 hours light, 8 hours dark
- Light intensity: Mean daylight intensity ranged from 82 to 87 footcandles.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : start of the test: 08:45; 24 h: 08.40, 13:00; 48 h: 09:00, 13:30; 72 h: 09:00, 13:00; 96 h: 13:00 (time)

TEST CONCENTRATIONS
- Range finding study: no

Reference substance (positive control):
no

Results and discussion

Effect concentrations
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
>= 1 mg/L
Nominal / measured:
estimated
Conc. based on:
test mat. (dissolved fraction)
Basis for effect:
mortality
Remarks on result:
not measured/tested
Details on results:
- Behavioural abnormalities: There were no behavioral or coloration abnormalities noted during this study.
- Observations on body length and weight: Control fish were individually weighed and measured in accordance to guidelines developed to insure consistency. Data were recorded on the appropriate form in the study data book (not included in study report).
- Mortality of control: None
- Abnormal responses: None
- Effect concentrations exceeding solubility of substance in test medium: The Total Carbon analyses for the stock solution batches used during this study indicated that the test material water soluble fraction could not be measured in solution.

Due to a technician error, the computer water quality study was not started prior to study initiation. As a result, that information was extracted from the daily computer reports, exact copies of which are included in the raw data book. No study related data were lost. A number of temperature alarms occurred during which the temperature exceeded protocol specifications. The mean temperature during the study is within protocol specifications and the excursions appear insignificant to the outcome of the study.

Applicant's summary and conclusion

Validity criteria fulfilled:
not specified
Remarks:
Test method was not given and therefore validity was not evaluated, however survival in the control was 100%.
Conclusions:
Based on the estimated LC50 >1 mg/L, the test material is considered as acutely toxic to fish according to Regulation (EC) No. 1272/2008 (CLP).
Executive summary:

There were no behavioural or colouration abnormalities noted during this study. There was no mortality observed during this study; as a result, no calculations were performed and no LC50 could be determined, but estimated. It was considered to be > 1 mg/L.