N-(2-ethylhexyl)-1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]naphthalen-2-amine

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:

10-21.12.2012

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 below

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

see below

Principles of method if other than guideline:

The study was performed without analytical determination of the test substance concentrations. For the analytical determination of the test substance concentration, no suitable analytical method was found. Therefore for all evaluations and results, nominal concentrations of the test substance were used.

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, saturated solutions of the test substance in dilution water were prepared by adding measured amounts of the test substance to the dilution water. The solutions were stirred for period of 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
- Common name: Guppy
- Length at study initiation: 2.0 to 3.0 cm

ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions (same as test or not): Yes. The fish were kept in all-glass aquaria in aerated drinking water.
- Type and amount of food during acclimation: Feed for aquarium fish
- Feeding frequency during acclimation: Once a day
- Health during acclimation (any mortality observed): Healthy animals

FEEDING DURING TEST
- Feeding during test: No

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

Content of (Ca + Mg) in dilution water: 2.46 mmol/L

Test temperature:

22 ± 2 °C

pH:

7.8 (pH of dilution water)

Dissolved oxygen:

9.1 mg/L

Conductivity:

0.174 mS/m

Nominal and measured concentrations:

- Nominal concentrations: 100, 60, 40, 20 and 10 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Beakers
- Material, size, headspace, fill volume: 1000 mL per beaker
- Aeration: No
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Standard dilution water was prepared just before the test by admixing 2.5 mL of stored solutions A, B, C and D in 1000 mL of deionised water. Solutions A to D contained 117.6 g CaCl2.2H2O, 49.3 g MgSO4.7H2O, 25.9 g NaHCO3 and 2.3 g KCl, respectively, each in 1 L of deionised water. The dilution water was saturated with oxygen by aeration at 20 - 24 °C.
- Intervals of water quality measurement: The pH, temperature and dissolved oxygen were measured every 24 hours.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: No data
- Light intensity: Daylight

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
The fish mortality was observed every 24 hours.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.5 to 2
- Range finding study: Yes
- Test concentrations: 100, 60, 40, 20 and 10 mg/L
- Results used to determine the conditions for the definitive study: No definitive study was carried out. Based on a lack of toxicity of the test substance in the preliminary test, the study was terminated. The concentration of 100 mg/L was included in the preliminary test with 7 fish per group; therefore the conditions of the limit test have been fulfilled.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No mortality was seen in any dose group throughout the study.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- LC50: 96 hour LC50 = 140 mg/L (95 % confidence limit: 90 to 190 mg/L)
- Other: The mortality of test organisms by the reference substance obtained from the most recent testing facility reference was in the appropriate range. The sensitivity of test species and proficiency of the laboratory in test performance was therefore verified.

Validity criteria fulfilled:

no

Remarks:

No mortality was seen in the control and the dissolved oxygen concentration remained at least 60 % of initial value. However it was not possible to verify the test substance concentrations.

Conclusions:

Under the conditions of this study, the 96 hour LC50 was determined to be >100 mg/L and the 96 hour NOEC was 100 mg/L.

Executive summary:

The test substance, Solvent Red 19E, was investigated for acute toxicity to fish (Poecilia reticulata) under GLP conditions.

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 and OECD Test Guideline No 203, Fish, Acute Toxicity Test, Adopted July 17, 1992.

In a preliminary study, 7 fish per concentration were exposed the test material at nominal concentrations of 10, 20, 40, 60 and 100 mg/L under static conditions.

The nominal concentrations were used for all evaluations and results. The study was performed without analytical determination of test substance concentrations because no suitable analytical method was found.

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

As no toxicity of the test substance was found in preliminary test, the study was terminated. The concentration of 100 mg/L included in the preliminary test was performed using 7 fish per group; therefore the conditions of the limit test have been fulfilled. It was demonstrated that the LC50 is higher than this concentration, thus no further testing is required.

Under the conditions of this study, the 96 hour LC50 was determined to be >100 mg/L and the 96 hour NOEC was 100 mg/L. The 96 hour LL100 was >100mg/L.

Description of key information

The 96 hour LC50 was determined to be >100 mg/L and the 96 hour NOEC was 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The test substance, Solvent Red 19E, was investigated for acute toxicity to fish (Poecilia reticulata) under GLP conditions.

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 and OECD Test Guideline No 203, Fish, Acute Toxicity Test, Adopted July 17, 1992.

In a preliminary study, 7 fish per concentration were exposed the test material at nominal concentrations of 10, 20, 40, 60 and 100 mg/L under static conditions.

The nominal concentrations were used for all evaluations and results. The study was performed without analytical determination of test substance concentrations because no suitable analytical method was found.

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

As no toxicity of the test substance was found in preliminary test, the study was terminated. The concentration of 100 mg/L included in the preliminary test was performed using 7 fish per group; therefore the conditions of the limit test have been fulfilled. It was demonstrated that the LC50 is higher than this concentration, thus no further testing is required.

Under the conditions of this study, the 96 hour LC50 was determined to be >100 mg/L and the 96 hour NOEC was 100 mg/L. The 96 hour LL100 was >100mg/L.