2-[(2-methoxy-4-nitrophenyl)azo]-N-(2-methoxyphenyl)-3-oxobutyramide

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:

2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study meets requirements of OECD Guideline 203 and GLP requirements

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

Test item analysis was technically not possible.

GLP compliance:

yes (incl. QA statement)

Remarks:

GLP according to German "Chemikaliengesetz"

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION OF TEST SOLUTION
Preparation of the saturated solution: A dispersion (100 mg/L test item was weighed out) was prepared with dilution water, shaken with 20 rpm for 48 h and centrifugated with 3000 upm for 20 minutes.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Source: Aquarium am Aegi, Volgersweg 6, D-30175 Hannover, Germany
- Length at study initiation (mean): 2.21 cm
- Weight at study initiation (mean): 0.4 g
- Method of breeding: Holding was performed at the testing facility at 23.2 °C and diffuse light (0.1-10 µmol photons * m-2 * s-1, natural photoperiod). The water was changed at least once per week. The dissolved oxygen concentration was more than 80 % of the air saturation value.

- Feeding during test
No feeding during the test. The test fish were not fed 24 h before the test started.

ACCLIMATION
- Acclimation period: 12 days of acclimatisation
- Acclimation conditions (same as test or not): Same as test conditions, but fish were fed.
- Type and amount of food: Stör perlets; SERA GMBH, D-52518 Heinsberg, Germany. The amount of food was 4 % of the fish body weight per feeding day.
- Feeding frequency: Food was provided 3 times per week.
- Health during acclimation (any mortality observed): Mortality <5%, no disease treatments were administered.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

Total hardness: 100 mg CaCO3/L (control & treatment)

Test temperature:

21.2°C - 22.0°C (control), 21.9°C - 22.7°C (treatment)

pH:

7.33 - 7.67 (control), 7.44 - 7.74 (treatment)

Dissolved oxygen:

97-100% Saturation

Nominal and measured concentrations:

Nominal concentration: 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Glass-aquaria, about 3 L per vessel, loosely covered by glass tops;
- Aeration: Gentle aeration was provided.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: fish density in the tanks was less than 1 g per litre test solution.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water of local origin was used for holding and testing. The water was filtered on activated charcoal and aerated for at least 24 h to remove chlorine. Nominal water parameters: pH-value: 6.0 - 8.5; The water is analysed biannually acc. to German tap water regulation (see above for actual data for control and treatment).
- Culture medium different from test medium: no
- Intervals of water quality measurement:
pH , temperature and oxygen saturation: at the beginning and every 24 h in all vessels;
Total hardness: at the beginning;

OTHER TEST CONDITIONS
- Photoperiod: natural photoperiod
- Light intensity: 0.1 - 10 µmol photons • m-2 • s-1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and behavior of the test fish and control fish were observed at 2, 24, 48, 72, 96 h.

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: solubility limit

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

other: > saturation level

Nominal / measured:

nominal

Basis for effect:

mortality (fish)

Remarks on result:

other: NOEC > saturated solution

Details on results:

- There was no mortality in control as well as in treated fish at any observation time point.
- Behavioural abnormalities: Neither in treated nor in control fish could any abnormalities be observed at any observation time points.

Sublethal observations / clinical signs:

- There was no mortality in control as well as in treated fish at any observation time point. - Behavioural abnormalities: Neither in treated nor in control fish could any abnormalities be observed at any observation time points.

Validity criteria fulfilled:

yes

Conclusions:

In this study Pigment Yellow 74 did not cause any effects to zebrafish after 96 h when tested with the saturated solution. The NOEC (0 – 96 h) is laid down as > concentration of the saturated solution.

Executive summary:

The acute toxicity of the test item Pigment Yellow 74 to fish (zebrafish) was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1992) and EC Commission Directive 92/69/EC Method C.1 (1992) from 2006-05-15 to 2006-05 -19 at DR.U.NOACK-LABORATORIEN, Sarstedt, Germany. A static limit test with a saturated solution was conducted. The preparation method has been chosen with regard to the test item properties and in accordance with the relevant OECD guidance document (2000). Duration of the test was 96 h. 7 test organisms were exposed to the saturated solution and the control. Water quality parameters pH-value, temperature and oxygen-saturation measured after 0, 24, 48, 72 and 96 h were determined to be within the acceptable limits. No test item analysis was carried according to the sponsor.

The saturated solution of the test item Pigment Yellow 74 didn´t show any signs of toxicity nor lethality at all observation time points up to 96 hours. Therefore, the NOEC was laid down as > the concentration of the saturated solution.

Description of key information

NOEC (96h) > saturated solution  for Danio rerio (OECD 203)

Additional information

The acute toxicity of the test item to fish (zebrafish) was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1992). A static limit test with a saturated solution was conducted. The preparation method has been chosen with regard to the test item properties and in accordance with the relevant OECD guidance document (2000).

Compared to the control, no signs of toxicity could be observed in Danio rerio with the saturated solution. Therefore, the NOEC (96h) was determined to be higher than the concentration of a saturated solution of Pigment Yellow 74 (>= 100 mg/l based on the nominal concentration).