Phenol, 4-(phenylamino)-, sulfurized, leuco deriv.

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:

09 October 2017 - 11 December 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

July 17, 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

31 May, 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test item solution used in the test was prepared by mechanical dispersion of 0.5 g test item into 5000 mL test medium (ISO medium), in order to obtain the concentration of 100 mg/L one day before the start of the study. After approx. 15 minutes ultrasonic dispersion the test item solution was stirred for about 24 hours.
- Controls: dilution water (ISO medium)

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish (Danio rerio)
- Source: Akvárium magazin Kft. (Pasaréti Gyula), 1222 Budapest, Dévény u. 36.
- Weight at study initiation: Mean control: 0.18 g, mean exposure group (100 mg/L): 0.23 g

ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions: same as test (12 to 16 hours photoperiod daily; 21 – 25°C)
- Type of food during acclimation: fish were fed with appropriate, commercial diet for fish
- Feeding frequency during acclimation: at least three times per week, until 24 hours before the test start
- Health during acclimation: No significant mortality (less than 5 % of population) occurred in seven days before the start of the experiment.

FEEDING DURING TEST
- The fish were not fed during the test.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

The fish were exposued continuously and observed at approximately 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality.

Hardness:

The reconstituted water (ISO medium) has an approximate theoretical total hardness of 249 mg/L (as CaCO3).

Test temperature:

The measured temperatures were within a range of 22.5 – 23.3°C in the test aquariums (the maximum deviation did not exceed ± 1°C).

pH:

The test was carried out without adjustment of pH. The measured pH values were in the range of 7.46 – 7.94 during the test.

Dissolved oxygen:

The dissolved oxygen concentration was in the range of 76.2– 94.8 % of the air saturation value at the temperature used. Test solutions were not aerated during the test.

Nominal and measured concentrations:

nominal: 100 mg/L, no measured concentrations

Details on test conditions:

TEST SYSTEM
- Aeration: Test solutions were not aerated during the test.
- No. of vessels per concentration (replicates): Ten fish were used as control (no replicates were used).
- No. of vessels per control (replicates): Ten fish were exposed to the test concentration (no replicates were used).

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO medium, prepared according to Annex 2 of the OECD guideline 203

OTHER TEST CONDITIONS
- Adjustment of pH: The test was carried out without adjustment of pH.
- Photoperiod: 16 hours light (artificial illumination) and 8 hours darkness

EFFECT PARAMETERS MEASURED: The fish were observed at approximately 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. The total length of each fish was measured at the end of the test.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: limit test (100 mg/L)
- Range finding study
- Test concentrations: 10, 50 and 100 mg/L (nominal concentrations)
- Results used to determine the conditions for the definitive study: Yes, no dead fishes observed in any of the dose groups after 96 hours.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr. (dissolved fraction)

Basis for effect:

mortality (fish)

Remarks:

or sub-lethal effects

Remarks on result:

other: There was no mortality or sub-lethal effects observed in 10 fish in the control group or in 10 fish exposed to a test concentration of 100 mg/L

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr. (dissolved fraction)

Basis for effect:

mortality (fish)

Remarks:

or sub-lethal effects

Remarks on result:

other: There was no mortality or sub-lethal effects observed in 10 fish in the control group or in 10 fish exposed to a test concentration of 100 mg/L.

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr. (dissolved fraction)

Basis for effect:

mortality (fish)

Remarks:

mortality or sublethal effects

Remarks on result:

other: There was no mortality or sub-lethal effects observed in 10 fish in the control group or in 10 fish exposed to a test concentration of 100 mg/L.

Details on results:

- Behavioural abnormalities: none
- Observations on body length and weight: none
- Other biological observations: none
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none
- Any observations: precipitation was observed

Sublethal observations / clinical signs:

Results of the PreliminaryRange-Finding Test (96-h)

Nominal concentrations [mg/L]	Control	100	50	10
Number of treated animals	3	3	3	3
Number of dead animals	0	0	0	0

 

Cumulative mortality data in the definitive test

Test Group	Cumulative mortality (initial population = 10 fish / test group)
	24h	48h	72h	96h
Control	0	0	0	0
100 mg/L	0	0	0	0

Note: No sublethal effects occurred throughout the test in the control and the treatment groups

 

Validity criteria fulfilled:

yes

Conclusions:

In this 96-hour acute toxicity test on Zebrafish (Danio rerio) the test item Leuco Sulfur Brown 96 had no toxic effect on fish up to a concentration of 100 mg/L. Accordingly, the 96-h LC50 value was determined to be > 100 mg/L and the 96-h NOEC was determined to be 100 mg/L, based on nominal test concentration.

Executive summary:

The acute toxicity of the test item to zebrafish was determined according to OECD Guideline 203. A limit test was performed using a single concentration of 100 mg test item/L. ISO medium was used as dilution water. Fish were exposed to aqueous test media containing the test item for 96 hours in a static test. Ten fish were exposed to the test concentration and ten fish were run as control (no replicates were used). Fish were observed at approximately 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. Measurements of pH, dissolved oxygen concentration and temperature were carried out daily.

There was no mortality or sub-lethal effects observed in fishes of the control group or in fishes exposed to a test concentration of 100 mg/L. The 96-h No Observed Effect Concentration (NOEC) was determined to be 100 mg/L and the LC50 and the LOEC values were determined to be > 100 mg/L, based on nominal test concentration.

Description of key information

In a 96-hour acute toxicity test on Zebrafish (Danio rerio) the test item Leuco Sulfur Brown 96 had no toxic effect on fish up to a concentration of 100 mg/L. Accordingly, the 96-h LC50 value was determined to be > 100 mg/L and the 96-h NOEC was determined to be 100 mg/L, based on nominal test concentration.

 

Additional information

The acute toxicity of the test item to zebrafish was determined according to OECD Guideline 203. A limit test was performed using a single concentration of 100 mg test item/L. ISO medium was used as dilution water. Fish were exposed to aqueous test media containing the test item for 96 hours in a static test. Ten fish were exposed to the test concentration and ten fish were run as control (no replicates were used). Fish were observed at approximately 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. Measurements of pH, dissolved oxygen concentration and temperature were carried out daily.

There was no mortality or sub-lethal effects observed in fishes of the control group or in fishes exposed to a test concentration of 100 mg/L. The 96-h No Observed Effect Concentration (NOEC) was determined to be 100 mg/L and the LC50 and the LOEC values were determined to be > 100 mg/L, based on nominal test concentration.