2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2-(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

experimental data of read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE report is based on two short term toxicity to fish studies as-
WoE 2. and WoE 3.

GLP compliance:

not specified

Analytical monitoring:

no

Remarks:

WoE 2 and WoE 3: no

Vehicle:

no

Remarks:

WoE 2 and WoE 3: no

Details on test solutions:

WoE 2: The test chemical concentrations taken foe the study were 0, 6.25 mg/L,12.5 mg/L, 25 mg/L, 50 mg/L,100 mg/L, respectively.
WoE 3: The test solution was prepared by dissolving 400 mg of the test chemical in 1 liters of potable water (passed through reverse osmosis system) with continuous stirring. This test solution was then added to the remaining three liters of water for achieving test concentrations of 100 mg/L.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

WoE 2:
TEST ORGANISM
- Common name: Zebra Fish
- Strain:Danio rerio
- Length at study initiation (length definition, mean, range and SD): average 1.7 cm
- Weight at study initiation (mean and range, SD): average 0.066 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

WoE 3:
TEST ORGANISM
- Common name:Zebra Fish
- Strain: Danio rerio
- Length at study initiation (length definition, mean, range and SD): average 3.42 cm
- Weight at study initiation (mean and range, SD): average 0.335 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: 12 hours light and 12 hours darkness
-Diet : Standard brand feed, once daily
-Acclimatization: seven days
-feeding frequency: Once in a day

Test type:

static

Water media type:

freshwater

Total exposure duration:

96 h

Hardness:

WoE 2: 150.5 mg of CaCO3
WoE 3: 140.5 mg of CaCO3

Test temperature:

WoE 2: 24°C
WoE 3: 25°C

pH:

WoE 2: 7.2
WoE 3: 7.14

Dissolved oxygen:

WoE 2: 6.9 mg/l
WoE 3: 6.66 mg/l

Nominal and measured concentrations:

WoE 2: Test chemical concentrations used for the study were 0, 6.25 mg/L,12.5 mg/L, 25 mg/L, 50 mg/L,100 mg/L, respectively.
WoE 3: Nominal concentration - 100 mg/l

Details on test conditions:

WoE 2 and WoE 3:
TEST SYSTEM
- Test vessel: Glass 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

Reference substance (positive control):

no

Remarks:

WoE 2 and WoE 3: no

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: WoE 2 and WoE 3

Sublethal observations / clinical signs:

WoE 2:

OBSERVATIONS:

Mortality and symptoms

Nominal Concentration (mg/l)	Number of fish		Mortality (cumulated) after
			3 h		6 h		24 h		48 h		72 h		96 h
			V1	V2	V1	V2	V1	V2	V1	V2	V1	V2	V1	V2
Control	4	4	-	-	-	-	-	-	-	-	-	-	-	-
6.25	4	4	-	-	-	-	-	-	-	-	-	-	-	-
12.5	4	4	-	-	-	-	-	-	-	-	-	-	-	-
25	4	4	-	-	-	-	-	-	-	-	-	-	-	-
50	4	4	-	-	-	-	-	-	-	-	-	-	-	-
100	4	4	-	-	-	-	-	-	-	-	-	-	-	-

 

Visible Symptoms

Nominal Concentration (mg/l)

Number of fish

symptom (cumulated) after

3 h

6 h

24 h

48 h

72 h

96 h

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

6.25

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

12.5

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

25

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

50

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

100

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

 

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

E: Exopthalamos

pH and oxygen content

Nominal Concentration (mg/l)	Number of fish		pH readings after
			24 h		48 h		72 h		96 h
			V1	V2	V1	V2	V1	V2	V1	V2
Control	4	4	7.4	7.4	7.5	7.5	7.7	7.7	7.8	7.8
6.25	4	4	7.6	7.6	7.8	7.8	7.9	7.9	7.9	7.9
12.5	4	4	7.8	7.8	7.8	7.8	8.0	8.0	8.1	8.1
25	4	4	7.8	7.8	7.9	7.9	8.1	8.1	8.2	8.2
50	4	4	7.9	7.9	8.0	8.0	8.1	8.1	8.2	8.2
100	4	4	7.9	7.9	8.0	8.0	8.1	8.1	8.2	8.2

 

Nominal Concentration (mg/l)	Number of fish		Oxygen content (mg/L) after
			24 h		48 h		72 h		96 h
			V1	V2	V1	V2	V1	V2	V1	V2
Control	4	4	6.9	6.9	6.8	6.8	6.5	6.5	6.4	6.4
6.25	4	4	6.6	6.6	6.4	6.4	6.2	6.2	6.1	6.1
12.5	4	4	6.6	6.6	6.4	6.4	6.2	6.2	6.1	6.1
25	4	4	6.5	6.5	6.4	6.4	6.2	6.2	6.0	6.0
50	4	4	6.5	6.5	6.4	6.4	6.1	6.1	6.0	6.0
100	4	4	6.5	6.5	6.4	6.4	6.1	6.1	6.0	6.0

 

Test temperature

Nominal Concentration (mg/l)	Number of fish		Temperature (°C) after
			24 h		48 h		72 h		96 h
			V1	V2	V1	V2	V1	V2	V1	V2
Control	4	4	23.2	23.2	24.1	24.1	23.7	23.7	23.5	23.5
6.25	4	4	23.2	23.2	24.1	24.1	23.7	23.7	23.5	23.5
12.5	4	4	23.2	23.2	24.1	24.1	23.7	23.7	23.5	23.5
25	4	4	23.2	23.2	24.1	24.1	23.7	23.7	23.5	23.5
50	4	4	23.2	23.2	24.1	24.1	23.7	23.7	23.5	23.5
100	4	4	23.2	23.2	24.1	24.1	23.7	23.7	23.5	23.5

WoE 3:

Mortality and symptoms:

Nominal Concentration (mg/l)	Number of fish		Mortality (cumulated) after
			3h		6h		24h		48h		72h		96h
			V1	V2	V1	V2	V1	V2	V1	V2	V1	V2	V1	V2
Control	4	4	-	-	-	-	-	-	-	-	-	-	-	-
100	4	4	-	-	-	-	-	-	-	-	-	-	-	-

Visible Symptoms:

Nominal Concentration (mg/l)

Number of fish

symptom (cumulated) after

3h

6h

24h

48h

72h

96h

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

100

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

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:

NominalConcentration (mg/l)	Number of fish		pH readings after
			24h			48h		72h		96h
			V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	7.26		7.27	7.41	7.46	8.11	8.12	8.17	8.19
100	4	4	7.71		7.22	7.81	7.82	7.96	7.97	8.13	8.19

 

NominalConcentration (mg/l)	Number of fish		Temperature (°C) after
			24h			48h		72h		96h
			V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	7.1		7.5	7.1	7.6	6.9	6.3	5.4	5.4
100	4	4	6.8		6.9	7.1	7.4	6.2	6.1	6.7	6.8

Test temperature

NominalConcentration (mg/l)	Number of fish		Oxygen content (mg/L) after
			24h			48h		72h		96h
			V1	V2		V1	V2	V1	V2	V1	V2
0 (control)	4	4	22		22	22	22	22	22	22	22
100	4	4	22		22	22	22	22	22	22	22

Validity criteria fulfilled:

not specified

Conclusions:

On the basis of the experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach, the 96 hr LC50 value of the test chemical on test fish can be expected to be > 100 mg/l.

Executive summary:

Data available for its structurally and functionally similar read across chemicals has been reviewed to determine the toxic effect of the test chemical on fish.The studies are as mentioned below:

An acute toxicity study was conducted for 96 hrs for assessing the effect of test chemical on fish. The test was performed following the OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average weight 0.066 g and average length of 1.7 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. The test conditions during the housing of the test organisms were oxygen content of 7.0 mg/l, pH 7.4, water temperature 24°C and under a photoperiod of 16:8 hr light: dark conditions, respectively. Test concentrations selected for the study was were 0, 6.25, 12.5, 25, 50 and 100 mg/L, respectively. Total 8 fishes were exposed to test chemical in a 5 lit bowl aquaria containing 4 liters of potable water. The test vessels were placed in a room at a temperature of 24°C, pH 7.2, hardness of water 150.5 mg of CaCO3 and under a photoperiod of 16:8 hr light: dark conditions, respectively. Aeration in test vessels was provided 1 day before the start of the experiment. No mortalities were observed in the control and test vessel. On the basis of effect of test chemical on mortality of the test organism, the median lethal concentration (LC50 (96 h)) value was determined to be > 100 mg/l.

Another short term toxicity to fish was conducted for 96 hrs for assessing the effect of test chemical. The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average weight 0.335 g and average length of 3.42 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. The test conditions during the housing of the test organisms were oxygen content of 7.06 mg/l, pH 7.23, water temperature 24°C and under a photoperiod of 12:12 hr light: dark conditions, respectively. Test concentrations selected for the study was 100 mg/L. Thus, limit test was performed using 100 mg/l of test chemical concentration. Total 8 fishes were exposed to test chemical in a 5 lit bowl aquaria containing 4 liters of potable water. The test vessels were placed in a room at a temperature of 25°C, pH 7.14, hardness of water 140.5 mg of CaCO3 and under a photoperiod of 12:12 hr light: dark conditions, respectively. Aeration in test vessels was provided 1 day before the start of the experiment. No mortalities were observed in the control and test vessel. On the basis of effect of test chemical on mortality of the test organism, the median lethal concentration (LC50 (96 h)) value was determined to be > 100 mg/l.

On the basis of the experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach, the 96 hr LC50 value of the test chemical on test fish can be expected to be > 100 mg/l. Thus, test chemical was considered as non-toxic to aquatic fishes and hence, considered to be not classified as per the CLP classification criteria.

Description of key information

On the basis of the experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach, the 96 hr LC50 value of the test chemical on test fish can be expected to be > 100 mg/l. 

Additional information

Data available for its structurally and functionally similar read across chemicals has been reviewed to determine the toxic effect of the test chemical on fish. The studies are as mentioned below:

 

An acute toxicity study was conducted for 96 hrs for assessing the effect of test chemical on fish. The test was performed following the OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average weight 0.066 g and average length of 1.7 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. The test conditions during the housing of the test organisms were oxygen content of 7.0 mg/l, pH 7.4, water temperature 24°C and under a photoperiod of 16:8 hr light: dark conditions, respectively. Test concentrations selected for the study was were 0, 6.25, 12.5, 25, 50 and 100 mg/L, respectively. Total 8 fishes were exposed to test chemical in a 5 lit bowl aquaria containing 4 liters of potable water. The test vessels were placed in a room at a temperature of 24°C, pH 7.2, hardness of water 150.5 mg of CaCO3 and under a photoperiod of 16:8 hr light: dark conditions, respectively. Aeration in test vessels was provided 1 day before the start of the experiment. No mortalities were observed in the control and test vessel. On the basis of effect of test chemical on mortality of the test organism, the median lethal concentration (LC50 (96 h)) value was determined to be > 100 mg/l.

 

Another short term toxicity to fish was conducted for 96 hrs for assessing the effect of test chemical. The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average weight 0.335 g and average length of 3.42 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. The test conditions during the housing of the test organisms were oxygen content of 7.06 mg/l, pH 7.23, water temperature 24°C and under a photoperiod of 12:12 hr light: dark conditions, respectively. Test concentrations selected for the study was 100 mg/L. Thus, limit test was performed using 100 mg/l of test chemical concentration. Total 8 fishes were exposed to test chemical in a 5 lit bowl aquaria containing 4 liters of potable water. The test vessels were placed in a room at a temperature of 25°C, pH 7.14, hardness of water 140.5 mg of CaCO3 and under a photoperiod of 12:12 hr light: dark conditions, respectively. Aeration in test vessels was provided 1 day before the start of the experiment. No mortalities were observed in the control and test vessel. On the basis of effect of test chemical on mortality of the test organism, the median lethal concentration (LC50 (96 h)) value was determined to be > 100 mg/l.

 

On the basis of the experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach, the 96 hr LC50 value of the test chemical on test fish can be expected to be > 100 mg/l. Thus, test chemical was considered as non-toxic to aquatic fishes and hence, considered to be not classified as per the CLP classification criteria.