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Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
5 September 1995 - 13 October 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
The study was performed according to OECD and EC guidelines.The analysis failed to provide evidence of exposure concentrations. However, the test solution was a turbid, white suspension which confirms the exposure to the test substance.
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
EEC Directive 92/69, 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
Stability under storage conditions: stable
Stability in water: not indicated
Solubility in water: <2.4 mg/L at 20 °C
Analytical monitoring:
yes
Details on sampling:
No analytical method was available or could be developed for this compound. Therefore, quantative analyses were performed on Al and Mg. Consequently, no stability of the test substance could be tested. The analyses were performed by: TNO Institute of Environmental Sciences in Delft, the Netherlands

- Concentrations: 0, 0 (with vehicle) and 100 mg/L
- Sampling method first test: according to schedule below
Frequency: at t=0 and at t=96 h
Volume: 25 mL from the approximate centre of the test vessels.
- Storage before sampling: samples were stored in a refrigerator until shipment to the TNO laboratories at Delft, the Netherlands. All samples were filtered through a 0.45 µm filter acidified before storage and subsequent shipment.
Vehicle:
yes
Remarks:
Cremophor EL
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: the test substance solution was prepared by mixing 798.8 mg of test substance with 801.16 mg Cremophor EL and adding this micture quantatively to 8 litres of ISO-medium providing a nominal concentration of 100 mg/L. This dispersion was stirred for ca. 66 hours before the start of the exposure. The final solution was a turbid, white suspension.
- Controls: one control containing test medium without test substance or other additives (0 mg/L) and, one control containing test medium without test substance but with vehicle
- Chemical name of vehicle: Cremophor EL, dispersant
- Concentration of vehicle in test medium: 100 mg/L
Test organisms (species):
Cyprinus carpio
Details on test organisms:
TEST ORGANISM
- Common name: carp
- Strain: Linnaeus, 1758
- Source: Zodiac, proefacc., ''De Haar Vissen'', L.U. Wageningen, the Netherlands
- Age at study initiation: not indicated, fish were F1 from a single parent-pair bred in UV-treated water
- Length at study initiation: 3.12 +/- 0.26 cm
- Weight at study initiation: 1.03 +/- 0.42 g

ACCLIMATION
- Acclimation period: 12 days after delivery
- Acclimation conditions: same as test conditions
- Frequency and type of food during acclimation: daily with Trouvit or Artemia
- Health during acclimation: mortality during the seven days prior to the start of the test < 5%

FEEDING DURING TEST
- Feeding: no feeding from 24 hours prior to the test and during the total test period.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
250 mg CaCO3/L
Test temperature:
Between 20.0 to 22.0 °C
pH:
Between 7.7 and 8.1 throughout the test.
Dissolved oxygen:
Between 9.1 and 9.9 mg O2/L throughout the test.
Nominal and measured concentrations:
Nominal concentrations: 0, 0 (with Cremophor EL) and 100 mg/L
Measured concentration in both controls and 100 mg/L: < 1.52 mg/L. The concentration of test substance was based on the amount of Al calculated as follows: Concentration Al * (588/54)
Details on test conditions:
TEST SYSTEM
- Test vessel: 15 litres, all-glass, open, fill volume: 8 litre
- Aeration: continuously
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: 0.9 g fish/litre

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO, formulated using Milli-Ro water
- Culture medium different from test medium: no
- Intervals of water quality measurement: once a week

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours daily

EFFECT PARAMETERS MEASURED: mortality after 2, 24, 48, 72 and 96 hours. In addition, Dissolved oxygen content and pH were measured daily in all vessels, beginning at day 0. Temperature of the medium was measured daily in one control vessel, beginning at day 0. And fish length and weight was measured in 10 fish of the batch used for the test, prior to the start of the test.

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: preliminary test showed that no effects were present for the supersaturated solution
- Range finding study concentrations: 100 mg/L supersaturated solution and 100 mg/L filtrate of a solution
- Results used to determine the conditions for the definitive study: no, the test substance batch used was not in agreement with that protocolled. Hence, these results are to be considered as not relevant.
Reference substance (positive control):
yes
Remarks:
pentachlorophenol
Key result
Duration:
96 h
Dose descriptor:
LC50
Basis for effect:
mortality (fish)
Remarks on result:
other: The 96h-LC50 was above the solubility limit of the test substance in test medium.
Details on results:
- No behavioural, biological or other abnormalities observed.
- Effect concentrations exceeding solubility of substance in test medium: yes
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Mortality: 100% after 24 hours at 0.18, 0.32, 0.56 and 1.0 mg/L
- LC50: 0.13 mg/L, 95%-CI(0.10, 0.18) mg/L
- Other: sensitivity of the fish from this batch are to be validated as rather high in the historical range.
Sublethal observations / clinical signs:

For more details on analytical and final results refer to the attached document. (CRL 137789 - Tables.docx)

Validity criteria fulfilled:
no
Remarks:
No evidence that the actual concentrations were maintained at more that 80% of the initial concentration due to low solubility of test substance.
Conclusions:
Under the conditions of this study, the 96h-LC50 for carp exposed to Alcamizer 5 was above the solubility limit of the test substance in test medium. Even though analysis of the test solution showed that no levels of Al or Mg higher than those in the controls were found, the turbid, white suspension proved that the fish were exposed to the test substance.
Executive summary:

A study was performed to assess the effect of Alcamizer 5 on the mortality of carp (Cyprinus carpio) after 96 hours of exposure. The study was conducted in accordance with OECD Guideline 203 and under GLP conditions.

A limit test was performed with an exposure concentration of 100 mg/L in a static system. 100 mg/L Cremophor EL was used to disperse the test substance in water. One control contained test medium without test substance or other additives, a second control contained test medium without test substance but with Cremophor EL. 7 Carps were exposed to the test substance for 96 hours in all-glass, open vessels. Mortality was recorded at 2 h, 24 h, 48 h, 72 h and 96 h.

Quantative analyis was performed on Al and Mg in samples taken from 0 and 100 mg/L and the Cremophor control to determine the test concentrations. Analysis showed that no levels of Al or Mg higher than those in the controls were found in the 100 mg/L solution. Measured concentrations of test substance, based on Al were calculated to be < 1.52 mg/L in both controls and 100 mg/L solution. Water solubility was determined to be < 2.4 mg/L in an earlier study. The 100 mg/L solution was a turbid, white suspension which proved that the fish were exposed to the test substance.

No effects were seen at a nominal concentration of 100 mg/L.

Under the conditions of this study, the 96h-LC50 for carp exposed to Alcamizer 5 was above the solubility limit of the test substance in test medium.

Description of key information

A study was performed to assess the effect of Alcamizer 5 on the mortality of carp (Cyprinus carpio) after 96 hours of exposure. The study was conducted in accordance with OECD Guideline 203 and under GLP conditions.

A limit test was performed with an exposure concentration of 100 mg/L in a static system. 100 mg/L Cremophor EL was used to disperse the test substance in water. One control contained test medium without test substance or other additives, a second control contained test medium without test substance but with Cremophor EL. 7 Carps were exposed to the test substance for 96 hours in all-glass, open vessels. Mortality was recorded at 2 h, 24 h, 48 h, 72 h and 96 h.

Quantative analyis was performed on Al and Mg in samples taken from 0 and 100 mg/L and the Cremophor control to determine the test concentrations. Analysis showed that no levels of Al or Mg higher than those in the controls were found in the 100 mg/L solution. Measured concentrations of test substance, based on Al were calculated to be < 1.52 mg/L in both controls and 100 mg/L solution. Water solubility was determined to be < 2.4 mg/L in an earlier study. The 100 mg/L solution was a turbid, white suspension which proved that the fish were exposed to the test substance.

No effects were seen at a nominal concentration of 100 mg/L.

Under the conditions of this study, the 96h-LC50 for carp exposed to Alcamizer 5 was above the solubility limit of the test substance in test medium.

Key value for chemical safety assessment

Additional information