N-ethyl-N-[2-[1-(2-methylpropoxy)ethoxy]ethyl]-4-(phenylazo)aniline

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:

26.11. - 21.12.2012

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:

yes

Remarks:

(see Overall remarks)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION OF TEST SOLUTION (especially for difficult test substances)
Since the test substance is poorly soluble in dilution water at concentrations needed for the test, the saturated solutions of the test substance in dilution water were prepared for preliminary test.
The saturated solutions were prepared by adding measured amounts of the test substance to the dilution water. The solutions were stirred for period 72 hours on a shaft stirrer and consequently filtered through 0.45 μm filter. These solutions were used for testing.

Test organisms (species):

Poecilia reticulata

Details on test organisms:

TEST ORGANISM
- Source: certified supplier EMPLA s.r.o., Hradec Králové (Czech Republic)
- Length at study initiation: 2.0 – 3.0 cm
- Method of breeding: The fish were kept in all-glass aquaria in aerated drinking water.
- Feeding during test: without feeding

ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: "Tropica” - feed for aquarium fish
- Feeding frequency: once a day
- Health during acclimation (any mortality observed): healthy animals

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Content of (Ca+Mg) in dilution water: 2.46 mmol.L-1

Test temperature:

22±2°C

pH:

7.8

Dissolved oxygen:

9.1 mg.L-1

Nominal and measured concentrations:

28, 20, 14, 10, 7,5 and 3.6 mg.L-1

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 28 mg/L

Nominal / measured:

nominal

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

>= 27.9 mg/L

Nominal / measured:

nominal

Key result

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

>= 19.2 mg/L

Nominal / measured:

nominal

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

>= 16.8 mg/L

Nominal / measured:

nominal

Key result

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

>= 28 mg/L

Nominal / measured:

nominal

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 5 mg/L

Nominal / measured:

nominal

Key result

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

>= 5 mg/L

Nominal / measured:

nominal

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 5 mg/L

Nominal / measured:

nominal

Key result

Duration:

24 h

Dose descriptor:

LC100

Effect conc.:

> 28 mg/L

Nominal / measured:

nominal

Key result

Duration:

48 h

Dose descriptor:

LC100

Effect conc.:

> 28 mg/L

Nominal / measured:

nominal

Key result

Duration:

72 h

Dose descriptor:

LC100

Effect conc.:

> 28 mg/L

Nominal / measured:

nominal

Key result

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

>= 28 mg/L

Nominal / measured:

nominal

Results with reference substance (positive control):

- Results with reference substance valid? yes The mortality of test organisms by the reference substance (96h-LC50), obtained from the last reference test meets the calculated range from the interlaboratory test. The sensitivity of test species and proficiency of the laboratory in test performance was proved.
- Reference test: 96 hour – LC50 = 140 mg.L-1 (95% confidence limit: 90 – 190 mg.L-1)

Sublethal observations / clinical signs:

Calculation of LC50 values

	LC50 mg·L-1	Confidence limit	Approach function
24 hours	>28	-	-
48 hours	27.9	22.6 – 33.3	Polynomial 3
72 hours	19.2	11.9 – 26.4	Probit
96 hours	16.8	12.1 – 21.4	Gryck-Haustein

Validity criteria fulfilled:

yes

Conclusions:

The nominal concentrations were used for all evaluations and results because study was performed without analytical determination of test substance concentrations (see table above Calculation of LC50 values)

Executive summary:

The test substance, Solvent Yellow 124, was tested for acute toxicity on fish Poecilia reticulata.

The test was performed according to method C.1 – Acute toxicity for fish, Council Regulation (EC) No. 440/2008, published in O.J.L 142, 2008.

Since the test substance is poorly soluble in dilution water at concentrations needed for the test, the saturated solutions of the test substance in dilution water were prepared for preliminary and full test. The saturated solutions were prepared by adding measured amounts of the test substance to the dilution water. The saturated solutions were stirred for period 72 hours on a shaft stirrer and consequently filtered through 0.45μm filter.

 

The preliminary test was performed in range of test substance nominal concentrations 10 – 100 mg•L-1. Based on toxicity of the test substance found in preliminary test, the full test was performed in appropriate concentration range.

The full test was performed in range of test substance nominal concentrations 3.6 - 28 mg•L-1. The nominal concentrations were used for all evaluations and results because study was performed without analytical determination of test substance concentrations. No suitable analytical method was found. The test was performed as static.

Test results:                                                        

24 hour – LC50 > 28 mg•L-1          (nominal concentration)

48 hour – LC50 = 27.9 mg•L-1      (nominal concentration) (95% confidence limit: 22.6 – 33.3 mg•L-1)

72 hour – LC50 = 19.2 mg•L-1      (nominal concentration) (95% confidence limit: 11.9 – 26.4 mg•L-1)

96 hour – LC50 = 16.8 mg•L-1      (nominal concentration) (95% confidence limit: 12.1 – 21.4 mg•L-1)

 

24 hour – NOEC = 28 mg•L-1         (nominal concentration)

48 hour – NOEC = 5 mg•L-1           (nominal concentration)

72 hour – NOEC = 5 mg•L-1           (nominal concentration)

96 hour – NOEC = 5 mg•L-1           (nominal concentration)

 

24 hour – LC100 > 28 mg•L-1       (nominal concentration)

48 hour – LC100 > 28 mg•L-1       (nominal concentration)

72 hour – LC100 > 28 mg•L-1       (nominal concentration)

96 hour – LC100 = 28 mg•L-1       (nominal concentration)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

16.8 mg/L

Additional information

The test substance, Solvent Yellow 124, was tested for acute toxicity on fish Poecilia reticulata.

The test was performed according to method C.1 – Acute toxicity for fish, Council Regulation (EC) No. 440/2008, published in O.J.L 142, 2008.

Since the test substance is poorly soluble in dilution water at concentrations neededfor the test, the saturated solutions of the test substance in dilution water were preparedfor preliminary and full test. The saturated solutions were prepared by adding measured amounts of the test substance to the dilution water. The saturated solutions were stirred for period 72 hours on a shaft stirrer and consequently filtered through 0.45μm filter.

 

The preliminary test was performed in range of test substance nominal concentrations10 – 100 mg•L-1. Based on toxicity of the test substance found in preliminary test,the full test was performed in appropriate concentration range.

The full test was performed in range of test substance nominal concentrations3.6 - 28 mg•L-1.The nominal concentrations were used for all evaluations and results because study was performed without analytical determination of test substance concentrations. No suitable analytical method was found.The test was performed as static.

Test results:                                                        

24 hour – LC50 > 28 mg•L-1          (nominal concentration)

48 hour – LC50 = 27.9 mg•L-1      (nominal concentration) (95% confidence limit: 22.6 – 33.3 mg•L-1)

72 hour – LC50 = 19.2 mg•L-1      (nominal concentration) (95% confidence limit: 11.9 – 26.4 mg•L-1)

96 hour – LC50 = 16.8 mg•L-1      (nominal concentration) (95% confidence limit: 12.1 – 21.4 mg•L-1)