[No public or meaningful name is available]

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

Study period:

Experiment start date - 17 May 2010; Experiment completion date - 09 June 2010; Study completion date - 21 July 2010.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identification: FAT 40851/A TE
Batch Number: TZ 5891 / BOP 02-09
Purity: 69.9 % all coloured components
Appearance: Orange powder
Expiry Date: July 31, 2014
Storage Conditions: At room temperature at about 20 °C

Analytical monitoring:

yes

Details on sampling:

For determination of the actual test item concentration in this semi-static test, duplicate samples were taken from the test medium and the control just before the start and at the end of each test medium renewal period

All samples were taken from the approximate center of each aquarium without mixing the test medium, and were stored deep-frozen (at about -20 °C) immediately after sampling. Based on a pre-experiment (without GLP) the test item was shown to be stable under the storage conditions.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
At the start of the test and before the test medium renewal, the test medium of nominal 110 mg/L was freshly prepared by completely dissolving 550 mg of the test item (dosed amounts: 550.4 and 550.1 mg) in 5 liters of test water using ultrasonic treatment for 10 minutes and stirring for 10 minutes at room temperature. The test medium was freshly prepared just before introduction of the fish (= start of the exposure and prior to the test medium renewal).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish (Brachydanio rerio)
- Source: breeding culture at Harlan Laboratories
- Length at study initiation (length definition, mean, range and SD):mean body length of the fish was 2.8 ± 0.10 cm (Mean ± SD) (10 fish)
- Weight at study initiation (mean and range, SD):mean body wet weight was 0.16 ± 0.02 g (Mean ± SD) (10 fish)

ACCLIMATION
- Acclimation period: Eight days to the test water and temperature
- Acclimation conditions (same as test or not): Test
- Type and amount of food: 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, 49304 Melle / Germany).
- Health during acclimation (any mortality observed): No medication was applied during holding and acclimatization. During holding and acclimatization, no fish died in the test fish batch and all fish were healthy.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

125 mg/L as CaCO3 (1.25 mmol/L)

Test temperature:

The water temperature in the test vessels was maintained at 21-22 °C. The test vessels were slightly aerated during the test period.

pH:

see table "pH in the treatments" below

Dissolved oxygen:

see table "Oxygen Concentration" below

Salinity:

see table "Test Water" below

Nominal and measured concentrations:

The nominal concentration tested in the present study with zebra fish was 110 mg FAT 40851/A.

Details on test conditions:

TEST SYSTEM
- Test vessel: One glass test vessel with 5 liters of test medium was used for each treatment
- Aeration: The test vessels were slightly aerated during the test period.
- Renewal rate of test solution (frequency/flow rate): After 48h
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.22g fish wet weight /Liter

TEST MEDIUM / WATER PARAMETERS
see table "test Water below"

OTHER TEST CONDITIONS
- The water temperature in the test vessels was maintained at 21-22 °C. The test vessels were slightly aerated during the test period.
- Adjustment of pH: none
- Photoperiod/Light intensity: A 16-hour light to 8-hour dark photoperiod, with a 30-minute transition period was used (light intensity during the light period was approximately within the range of 140 to 480 Lux).

EFFECT PARAMETERS MEASURED : Mortality and Visible Abnormalities

TEST CONCENTRATIONS
The EC50 values for FAT 40851/A TE obtained in an algal and daphnia toxicity test (C67357 and C67346) were above the nominal value of 100 mg/L. Based on this, the nominal concentration tested in the present study with zebra fish was 110 mg FAT 40851/A TE.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 110 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 110 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Other biological observations: No visible observations
- Mortality of control: None

Sublethal observations / clinical signs:

Mortality and Visible Abnormalities Observed in the Test Fish

Nominal test item concentration (mg/L)	Number of abnormal and dead fish / number of dead fish Type of visible abnormalities
	Observation time
	4 hours	24 hours	48 hours	72 hours	96 hours
Control	0 / 0	0 / 0	0 / 0	0 / 0	0 / 0
110	0 / 0	0 / 0	0 / 0	0 / 0	0 / 0
LC50	>110 mg/L

Validity criteria fulfilled:

yes

Conclusions:

In conclusion, the test item FAT 40851/A had no acute toxic effects on zebra fish under the present test conditions at a nominal concentration of 110 mg/L. The 96 EC50 was calculated to be >110 mg/L.

Executive summary:

The acute toxicity of the test item FAT 40851/A to zebra fish (Brachydanio rerio) was determined in a 96-hour semi-static test with one test medium renewal after 48 hours according to the EU Commission Directive 92/69/EEC, Part C.1, the Commission Regulation (EC) No 440/2008, Part C.1 and the OECD Guideline for Testing of Chemicals No. 203 (1992). A limit test was performed in accordance with the threshold approach requiring that the fish test be performed with a single concentration close to or slightly higher than the lowest EC50 value obtained in tests with algae and daphnia. The threshold approach was used to demonstrate, that zebra fish is not the most sensitive species for FAT 40851/A . The EC50 values for FAT 40851/A obtained in an algal and daphnia toxicity test were above the nominal value of 100 mg/L. Based on this result, the nominal concentration tested in the present study with zebra fish was 110 mg FAT 40851/A. Additionally, a control was tested in parallel. In the analysed test medium samples from the start and end of the test medium renewal periods the measured test item concentration ranged from 88 to 95 % of the nominal value. The mean measured test concentration (calculated as arithmetic mean of all measurements) was 101 mg/L. The test water renewal ensured a constant concentration of the test item during the test and the reported biological results are related to the nominal test item concentration of 110 mg/L. In the control and at the test concentration of 110 mg/L no mortality or other visible abnormalities were determined at the test fish during the test period of 96 hours. In conclusion, the test item FAT 40851/A had no acute toxic effects on zebra fish under the present test conditions at a nominal concentration of 110 mg/L. The 96 EC50 was calculated to be >110 mg/L.

Description of key information

The substance was not acutely toxic to zebra fish in a semi-static aquatic freshwater toxicity test performed for 96 hours, where the LC50 >110 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

> 110 mg/L

Additional information

The acute toxicity of the test item FAT 40851/A to zebra fish (Brachydanio rerio) was determined in a 96-hour semi-static test with one test medium renewal after 48 hours according to the EU Commission Directive 92/69/EEC, Part C.1, the Commission Regulation (EC) No 440/2008, Part C.1 and the OECD Guideline for Testing of Chemicals No. 203 (1992). A limit test was performed in accordance with the threshold approach requiring that the fish test be performed with a single concentration close to or slightly higher than the lowest EC50 value obtained in tests with algae and daphnia. The threshold approach was used to demonstrate, that zebra fish is not the most sensitive species for FAT 40851/A. The EC50 values for FAT 40851/A obtained in an algal and daphnia toxicity test were above the nominal value of 100 mg/L. Based on this result, the nominal concentration tested in the present study with zebra fish was 110 mg FAT 40851/A. Additionally, a control was tested in parallel. In the analysed test medium samples from the start and end of the test medium renewal periods the measured test item concentration ranged from 88 to 95 % of the nominal value. The mean measured test concentration (calculated as arithmetic mean of all measurements) was 101 mg/L. The test water renewal ensured a constant concentration of the test item during the test and the reported biological results are related to the nominal test item concentration of 110 mg/L. In the control and at the test concentration of 110 mg/L no mortality or other visible abnormalities were determined at the test fish during the test period of 96 hours. In conclusion, the test item FAT 40851/A had no acute toxic effects on zebra fish under the present test conditions at a nominal concentration of 110 mg/L. The 96 EC50 was calculated to be >110 mg/L.