Disodium 6-hydroxy-5-[(2-methoxy-4-sulphonato-m-tolyl)azo]naphthalene-2-sulphonate

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Experimental test result performed according to the guideline.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with continuous stirring for achieving test concentrations of 100mg/L,respectively.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name:Zebra Fish
- Strain:Danio rerio
- Source: Anytime Pet,Jaripatka, Nagpur
- Length at study initiation (length definition, mean, range and SD): average 1.8 cm
- Weight at study initiation (mean and range, SD): average 0.082 g
-Housing:The fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration.
-Duration of housing: 15 days
-Photoperiod: 16 hours light and 8 hours dark
-Diet : Standard brand feed, once daily
-Acclimatization: seven days
-feeding frequency: Once in a day

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

155.5 mg of CaCO3

Test temperature:

24.1 °C

pH:

7.1

Dissolved oxygen:

7.0 mg/L

Nominal and measured concentrations:

100 mg/l

Details on test conditions:

TEST SYSTEM:
- Test vessel: Plastic aquaria
- Material, size, headspace, fill volume: 5 liters of plastic aquaria filled with 2 liter of water
- Aeration: Aeration in test vessels was provided 1 day before the start of experiment
- No. of organisms per vessel: 8 fishes

OTHER TEST CONDITIONS:
- Photoperiod:16 hours light and 8 hours dark
- Withdrawal of food: 1 day before and during exposure

Reference substance (positive control):

no

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)

Remarks on result:

other: No mortality were observed

Sublethal observations / clinical signs:

Mortality and symptoms:

Nominal conc. (mg/l)	Number of fish	Mortality (cumulated) after
		24h	48h	72h	96h
0 (control)	8	0	0	0	0
100	8	0	0	0	0

Visible Symptoms:

Nominal conc. (mg/l)	Number of fish	Symptoms after
		24h	48h	72h	96h
0 (control)	8	NS8	NS8	NS8	NS8
100	8	NS8	NS8	NS8	NS8

Explanation of symptoms:

A: apathy

N: Narcotic-like state

C: Swimming near water surface

M: Slow movements compared to control

NS: No abnormal symptoms

S: Surfacing behaviour

L: Loss of equilibrium

SM: Shimming movement

H1: Hyperactivity

H2: Hypo activity

B: Bending behavior

F: Fin rot

IS: Invert Swimming

SB: Swimming behaviour

RF: Respiratory Function

P: Pigmentation

pH and oxygen content:

Nominal conc. (mg/l)	pH readings after
	24h	48h	72h	96h
0 (control)	6.8	7.4	6.7	7.3
100	6.4	6.3	6.1	6.8

 

Nominal conc. (mg/l)	Oxygen content (mg/l) after
	24h	48h	72h	96h
0 (control)	4.6	6.9	6.7	6.2
100	6.5	5.03	4.4	6.7

Test temperature

Nominal conc. (mg/l)	Temperature (°C) after
	24h	48h	72h	96h
0 (control)	35	36	35	35
100	35	35	34	34

Validity criteria fulfilled:

yes

Conclusions:

Based on nominal concentration, experimental median lethal Concentrations [LC-50 (96 h)] for test material on Zebra Fish (Danio rerio) was determine to be >100 mg/l.

Executive summary:

Study was conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test) to assess the effect of test chemical on the mortality of fish Danio rerio. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with continuous stirring for achieving test concentrations of 100mg/L, respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to test item to 100 mg/l nominal test concentrations, LC50 was determine to be > 100 mg/l. Thus, based on the LC50, it was considered that the chemical was not toxic and cannot be classified as per the CLP classification criteria.

Description of key information

Based on nominal concentration, experimental median lethal Concentrations [LC-50 (96 h)] for test material on Zebra Fish (Danio rerio) was determine to be >100 mg/l.  

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

Based on the experimental data for the target as well as read-across analogues which are extracted by using mechanistic approach and functionally and structurally similar to the target chemical, toxicity of test chemical were determined. The studies are summarized as below:

 Study was conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test) to assess the effect of test chemical on the mortality of fish Danio rerio. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 400 mg of the test substance in 4 liters of potable water (passed through reverse osmosis system) with continuous stirring for achieving test concentrations of 100mg/L, respectively and Zebra Fish Danio rerio were exposed to these concentration for 96 hours. Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to test item to 100 mg/l nominal test concentrations, LC50 was determine to be > 100 mg/l. Thus, based on the LC50, it was considered that the chemical was not toxic and cannot be classified as per the CLP classification criteria.

 In a supportive evidence, a 96 h study was conducted to determine the effect of test chemical on fish Danio rerio, at a test concentrations of 6.25mg/L, 12.5mg/L, 25mg/L, 50mg/L, 100mg/L. Test was performed according to OECD Guideline 203 (Fish, Acute Toxicity Test). Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. Zebra Fish (Danio rerio) were exposed to this concentration for 96 hours. After 96 hours of exposure of test chemical at of 100mg/L, no mortality was observed. Therefore the LC50 was determined to be >100mg/L. Based on the LC50, the test chemical can be considered as non-toxic and can be concluded as not classified as per the CLP classification criteria.

 Further to support first and second study again, A Fish Acute Toxicity test was performed to evaluate the toxic nature of test substance. Test was performed according to OECD Guideline 203. Potable water (passed through reverse osmosis system) was used for the preparation of test solution The test was performed on the limit concentration i.e nominal concentration selected for the experiment were 100 mg/L and test fish were exposed to these concentration for 96 hours.

Effect on the symptoms and the normal activity was checked in the interval of 24rs. pH, tempterature and dissolved oxygen was also checked. After 96hrs of exposure, the lethal concentration LC50 was determined to be >100 mg/L

Thus based on the LC50 it can be concluded that the chemical was nontoxic and was considered to be not classified as per the CLP classification criteria.

 

Hence from the above data, the test chemical was considered as nontoxic and was not classified as per the CLP classification criteria.