Reaction mass ofDisodium [1-{[2-(hydroxy-kO)-3,5-dinitrophenyl]diazenyl-kN1}naphthalen-2-olato(2-)-kO][3-(hydroxy-kO)-4-{[2-(hydroxy-kO)naphthalen-1-yl]diazenyl-kN1}-7-nitronaphthalene-1-sulfonato(3-)]chromate(2-) andDisodium [1-{[2-(hydroxy-kO)-3,5-dinitrophenyl]diazenyl-kN2}naphthalen-2-olato(2-)-kO][3-(hydroxy-kO)-4-{[2-(hydroxy-kO)naphthalen-1-yl]diazenyl-kN1}-7-nitronaphthalene-1-sulfonato(3-)]chromate(2-)

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:

Experimental starting date - 11 May 1998; Experiment and study completion date - 15 May, 1998

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

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Code number: FAT 20037/D
- Source and batch No.of test material: EN 101495.CB
- Expiration date of the batch: March, 2003

OTHER SPECIFICS:
- Purity: ca. 79 %,
- Appearance: black, powder
- Solubility: 40 g/L
- Storage: room temperature

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

- Preparation of stock solution: 1000 mg test substance was dissolved in 200 mL dechlorinated tap water, was homogenized for 10 minutes by ultrasonic treatment and then added up to 1.0 litre. For the preparation of the test concentrations the stock solution was mixed.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra-fish
- Source: River Aquatics, Someren, Netherlands

ACCLIMATION
- Acclimation period: The quality of the fish was checked with an acclimatization time of 61 days in dechlorinated tap water.
- Feeding during the testing: The fish were not fed during the test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

142 mg/L CaCO3

pH:

7.9-8.4

Dissolved oxygen:

99 %

Nominal and measured concentrations:

Nominal test concentrations: 0, 0.08, 0.18, 0.40, 0.88, 1.9 and 4.3 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: Beaker
- Material, size, headspace, fill volume: 5 litre glass aquaria with a test volume of 3 litre
- Aeration: Slow aeration
- No. of organisms per vessel: 7

EFFECT PARAMETERS MEASURED: Mortality and behaviour of fish were observed and recorded at 24, 48, 72 and 96 hours

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.66 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 0.41 - 1.07 mg/L

Details on results:

Swimming behaviour was observed at concentrationns above 0.4 mg/L.

Sublethal observations / clinical signs:

In the test concentrations up to 0.40 mg/L no alteration of the test substance in the test media was observed during the test. In the concentrations of 0.88 up to 4.3 mg/L a small sediment at the bottom of the test vessels was observed after 24 hours. In the control and in the test concentrations up to 0.18 mg/L no mortalities and no abnormal behaviour of the test fish was observed over the test period. Therefore, the NOEC and the LC0 value was determined to be 0.18 mg/L. The LC100 value was determined to be 1.9 mg/L nominal.

Mortalities of fish

Nominal Concentration (mg/L)	Test duration
	24h	48h	72h	96h
Blank	0	0	0	0
0.08	0	0	0	0
0.18	0	0	0	0
0.4	1	2	2	2
0.88	2	4	4	4
1.9	7	-	-	-
4.3	7	-	-	-

Validity criteria fulfilled:

yes

Conclusions:

The LC50 value to fish was determined to be 0.66 mg/L nominal.

Executive summary:

An acute study was conducted according to OECD test guideline 203 and EU method C.1 to investigate the toxicity potential of the test item to the zebrafish species. FAT 20037/D was tested at nominal concentrations of 0.08, 0.18, 0.40, 0.88, 1.9 and 4.3 mg test substance/L. Additionally a blank control without test substance was also tested. For each concentration, 7 fish were used. The fish were not fed during the test. In the test concentrations up to 0.40 mg/L, no alteration of the test substance in the test media was observed during the test. In the concentrations of 0.88 up to 4.3 mg/L, a small sediment at the bottom of the test vessels was observed after 24 hours. No mortality was observed in control, 0.08 and 0.18 mg/L concentration. 2/7 and 4/7, fish were found dead at 0.4 and 0.88 mg/L respectively. No fish survived at 1.9 and 4.3 mg/L concentration. Based on the findings of the study, and using the Probit model method, the LC50 value to fish was determined to be 0.66 mg/L nominal.

Description of key information

The LC50 value for the test substance ranges from 0.66 to 2.3 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.66 mg/L

Additional information

Three studies are available for evaluation of toxicity potential of the test substance to fish.

The key acute study was conducted according to OECD test guideline 203 and EU method C.1 to investigate the toxicity potential of the test item to the zebrafish species. FAT 20037/D was tested at nominal concentrations of 0.08, 0.18, 0.40, 0.88, 1.9 and 4.3 mg test substance/L. Additionally a blank control without test substance was also tested. For each concentration, 7 fish were used. The fish were not fed during the test. In the test concentrations up to 0.40 mg/L, no alteration of the test substance in the test media was observed during the test. In the concentrations of 0.88 up to 4.3 mg/L, a small sediment at the bottom of the test vessels was observed after 24 hours. No mortality was observed in control, 0.08 and 0.18 mg/L concentration. 2/7 and 4/7, fish were found dead at 0.4 and 0.88 mg/L respectively. No fish survived at 1.9 and 4.3 mg/L concentration. Based on the findings of the study, and using the Probit model method, the LC50 value to fish was determined to be 0.66 mg/L nominal.

In a supporting acute fish toxicity study, three groups containing 8 fish each, were exposed to test item concentrations of 0.10, 1.0 and 3.0 mg/L. Observations of mortality were made at 24 and 48 hours during the exposure. No mortality was observed at the concentrations of 0.1 and 1.0 mg/L. However, 7 out of 8 fish died at the concentration of 3.0 mg/L. Hence, the LC50 was calculated to be 1.85 mg/L.

In another supporting acute fish toxicity study conducted on FAT 20037/A, four groups containing 8 fish each, were exposed to test item concentrations of 0.20, 2.0, 20.0 and 100.0 mg/L. Observations of mortality were made at 24 and 48 hours during the exposure. No mortality was observed at the concentration of 0.2 mg/L. At the test concentration of 2.0 mg/L, one fish was found dead at the 48 hours observation. However, 8 out of 8 fish died at the concentrations of 20.0 and 100.0 mg/L. Hence, the LC50 was calculated to be 2.3 mg/L.