Producto P0310

Short-term toxicity to fish

Link to relevant study record(s)

Reference

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:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

For the analysis of the actual test item concentrations the following samples were taken
just before test start:
- duplicate samples from the test medium
- duplicate samples from the control
after 48 and 96 hours:
- duplicate samples from the test medium (stability samples)
- duplicate samples from the control
All samples were taken from the approximate center of the aquaria without mixing the test medium, and were deep-frozen (at about -20 ºC) immediately after sampling. Based on pre-experiments for investigation of the storage stability the test item is sufficiently stable in the test water under the storage conditions. The concentration of the test item was analytically measured in the duplicate test medium samples of the single test concentration taken at the start and the end of the test. From the control, one of the duplicate samples was analyzed from each of the sampling times.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: At the start of the test, the test medium of nominal 100 mg/L was freshly prepared by completely dissolving 500.9 mg of the test item in five liters of test water using ultrasonic treatment for 15 minutes and stirring for 15 minutes at room temperature in the dark. The test medium was freshly prepared just before introduction of the fish (= start of the test).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Length at study initiation (length definition, mean, range and SD): 3.1 ± 0.1 cm
- Weight at study initiation (mean and range, SD): 0.59 ± 0.02 g
- Feeding during test: No
- Food type: During holding and acclimatization until one day before the start of the test the fish were fed with a commercial fish diet (Tetra Min® Hauptfutter, supplied by TETRA-Werke, D-49304 Melle, Germany).


ACCLIMATION
- Acclimation period: One week

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

Water Hardness : 1.25 mmol/L (= 125 mg/L as CaCO3)

Test temperature:

Water temperature: 22 ºC during the test period

pH:

The pH values in the test medium and the control were between 7.3 and 7.4.

Dissolved oxygen:

The oxygen concentration was always 8.5 mg/L or higher, and thus higher than 60% oxygen saturation.

Salinity:

Reconstituted water: analytical grade salts were dissolved in deionized water to obtain the following nominal concentrations:
CaCl2 × 2H2O : 1.00 mmol/L (= 147 mg/L)
MgSO4 × 7H2O : 0.25 mmol/L (= 61.5 mg/L)
NaHCO3 : 0.38 mmol/L (= 32.5 mg/L)
KCl : 0.038 mmol/L (= 2.9 mg/L)

Nominal and measured concentrations:

Nominal concentration of 100 mg/L..
Mean measured test item concentration of 93 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: One glass aquarium with 5 liters test medium was used for each treatment (the single test concentration and the control).
- No. of organisms per vessel: 7 fish
- No. of vessels per concentration (replicates): One
- No. of vessels per control (replicates): One

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water: analytical grade salts were dissolved in deionized water
- Intervals of water quality measurement: The water temperature, pH-values and dissolved oxygen concentrations were measured at the start of the test and once every day during the test in the single test concentration and in the control.

OTHER TEST CONDITIONS
- Photoperiod: Since the test item is not stable under light conditions, the test was performed as far as possible in the dark to avoid photolytic degradation of the test item.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The test fish were observed after approximately 2, 24, 48, 72 and 96 hours test duration for mortality and visible abnormalities.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Basis for effect:

mortality (fish)

Remarks:

All fishes survived.

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

>= 93 mg/L

Nominal / measured:

meas. (arithm. mean)

Basis for effect:

mortality (fish)

Remarks:

All fishes survived.

Details on results:

In the control and in the test medium with the nominal concentration of 100 mg/L (mean measured concentration of 93 mg/L) all fish survived until the end of the test and no visible abnormalities were observed at the test fish.

Sublethal observations / clinical signs:

At the start of the test, the analytically determined test item concentration in the analyzed test medium was 94% of the nominal value, proving a correct dosage of the test item. During the test period of 96 hours, a slight decrease of the test item concentration in the test medium was determined. At the end of the test, 90% of the nominal value was found. Photolytic degradation could be minimized by reduction of the light intensity. The mean measured test concentration (calculated as the geometric mean over all measurements of the test concentration) was 93 mg/L (i.e., 93% of the nominal value).

Validity criteria fulfilled:

yes

Remarks:

(mortality in control was 0%; constant conditions were maintained; the dissolved oxygen concentration was within the the acceptable range; the concentrations were maintained during the test).

Conclusions:

The 96-hour NOEC and the 96-hour LC0 of the test item to zebra fish were determined to be at least the nominal concentration of 100 mg/L (mean measured concentration of 93 mg/L). The 96-hour NOEC and the 96-hour LC0 might even be higher than this concentration, but nominal concentrations in excess of 100 mg/L have not been tested, according to the guidelines.

Executive summary:

The acute toxicity of the test item to zebra fish (Brachydanio rerio) was determined according to the EU Commission Directive 92/69/EEC, Part C.1 (1992), and the OECD Guideline for Testing of Chemicals No. 203 (1992). A limit test was performed in accordance with the guidelines to demonstrate that the test item has no toxic effect on the test organisms at the nominal concentration of 100 mg/L. Thus, a single test item concentration of nominal 100 mg/L was tested. Additionally, a control was tested in parallel (test water without addition of the test item). At the start of the test, the analytically determined test item concentration in the analyzed test medium was 94% of the nominal value, proving a correct dosage of the test item. During the test period of 96 hours, a slight decrease of the test item concentration in the test medium was determined. At the end of the test, 90% of the nominal value was found. Photolytic degradation could be minimized by reduction of the light intensity. The mean measured test concentration was 93 mg/L. The reported biological results are related both to the nominal and to the mean measured test item concentrations. In the control and in the test medium with the nominal concentration of 100 mg/L (mean measured concentration of 93 mg/L) no mortality or other visible abnormalities were determined at the test fish during the test period of 96 hours. Therefore, the 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours), and the 96-hour LC0 of the test item to zebra fish were determined to be at least the nominal concentration of 100 mg/L (mean measured concentration of 93 mg/L). The 96-hour NOEC and the 96-hour LC0 might even be higher than this concentration, but nominal concentrations in excess of 100 mg/L have not been tested, according to the guidelines.

Description of key information

Key study: OECD guideline 203 and EU method C.1. GLP study.

The 96-hour NOEC and the 96-hour LC0 of the test item to zebra fish were determined to be at least the nominal concentration of 100 mg/L (mean measured concentration of 93 mg/L). The 96-hour NOEC and the 96-hour LC0 might even be higher than this concentration, but nominal concentrations in excess of 100 mg/L have not been tested, according to the guidelines.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

Key study: OECD guideline 203 and EU method C.1. GLP study.

The 96-hour NOEC and the 96-hour LC0 of the test item to zebra fish were determined to be at least the nominal concentration of 100 mg/L (mean measured concentration of 93 mg/L). The 96-hour NOEC and the 96-hour LC0 might even be higher than this concentration, but nominal concentrations in excess of 100 mg/L have not been tested, according to the guidelines.