Disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Experimental test result performed using standard OECD test guidelines

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Principles of method if other than guideline:

This study was designed to assess the effect of the test item disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate (CAS No. 4553-89-3) on Danio rerio. The study was conducted in accordance with OECD Guideline 203 (Fish, Acute Toxicity Test).

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material: disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate
- Molecular formula: C27H18N4Na2O9S2
- Molecular weight: 652.57 g/mol
- Smiles notation: c12c(c(\N=N\c3c(c(\N=N\c4c5c(c(S(=O)(=O)[O-])cc4)cccc5)cc(c3O)CO)O)ccc1S(=O)(=O)[O-])cccc2.[Na+].[Na+]
- InChI: 1S/C27H20N4O9S2.2Na/c32-14-15-13-22(30-28-20-9-11-23(41(35,36)37)18-7-3-1-5-16(18)20)27(34)25(26(15)33)31-29-21-10-12-24(42(38,39)40)19-8-4-2-6-17(19)21;;/h1-13,32-34H,14H2,(H,35,36,37)(H,38,39,40);;/q;2*+1/p-2/b30-28+,31-29+;;
- Substance type: Organic
- Physical state: Solid
- Batch No.: FG/14-15/1097
- Physical Appearance: Dark brown powder
- Purity: 79.4 % (dye content by GC)
- Stability: Stable in recommended storage conditions
- Storage Conditions: 1. Ambient Temp (23 to 27 °C), 2. In Refrigerator (4 °C), 3. In Freezer (-10 To 0 °C)
- Solubility in water: 99418 mg/L

Analytical monitoring:

not specified

Vehicle:

no

Details on test solutions:

The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 1 gm of the test substance in 1 liter of potable water (passed through reverse osmosis system) with continuous stirring for achieving test concentrations of 6.25 mg/L, 12.5mg/L,25mg/L, 50mg/L, 100mg/L, respectively

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra Fish (Danio rerio)
- Source: Asim Fancy Fish Aquarium, Dharampeth, Nagpur, MS (India)
- Length at study initiation (length definition, mean, range and SD):average 2.46 cm
- Weight at study initiation (mean and range, SD):0.0942 g (approx.)
- Physical conditions: No abnormalities

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

62.5mg of CaCO3

Test temperature:

22°C

pH:

7.3

Dissolved oxygen:

7.9 mg/L

Nominal and measured concentrations:

6.25 mg/L, 12.5mg/L,25mg/L, 50mg/L, 100mg/L

Details on test conditions:

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


OTHER TEST CONDITIONS
- Adjustment of pH: 7.3
- Photoperiod: 12 hours light and 12-hour darkness
- TDS (ppm): 245
- Conductivity (µScm-1): 658

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)

Details on results:

Determination or calculation of the median lethal concentrations (LC 50) and observations of the symptoms were done after 24, 48, 72 and 96 hours.
LC0 (96 hours) (highest loading at which no mortality was observed) = 100mg/L
LC50 (96 hours) Experimental = > 100mg/L
LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 100% mortality was not observed in any of the concentrations

Sublethal observations / clinical signs:

Mortality and symptoms

Nominal Concentration (mg/l)	Number of fish		Mortality (cumulated) after
			3 h	6 h	24h			48h		72h		96h
			V1	V2	V1	V2		V1	V2	V1	V2	V1	V2
0 (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

Symptoms after

3 h

6 h

24h

48h

72h

96h

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

0 (control)

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

6.25

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

12.5

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

25

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

50

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

100

4

4

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

Validity criteria fulfilled:

yes

Conclusions:

A 96 h study was conducted for CAS No. 4553-89-3 on Zebra Fish (Danio rerio), at a test concentrations of 6.25 mg/L, 12.5mg/L,25mg/L, 50mg/L, 100mg/L. The fishes were moving slowly as compared to control. No mortalities were found in the control aquaria. The lethal concentrations LC50 value was determined to be > 100mg/L.

Executive summary:

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for Disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 1 gm of the test substance in 1 liter of potable water (passed through reverse osmosis system). The nominal concentration selected for the experiment were 6.25 mg/L, 12.5mg/L, 25mg/L, 50mg/L, 100mg/L and Zebra Fish (Danio rerio) were exposed to these concentration for 96 hours. The fishes were moving slowly as compared to control. No mortalities were found in the control aquaria.  The lethal concentrations LC50 value was determined to be > 100mg/L. Based on the above LC50 value it can be concluded that the target substance 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate is non toxic to fish and thus can not be classified under Aquatic toxic category.

Description of key information

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for Disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate

(UERL study report, Report no. 4553 -89 -3/01/2015/FT, Sustainability Support Services (Europe) AB has the letter of access, 2015). The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 1 gm of the test substance in 1 liter of potable water (passed through reverse osmosis system). The nominal concentration selected for the experiment were 6.25 mg/L, 12.5mg/L, 25mg/L, 50mg/L, 100mg/L and Zebra Fish (Danio rerio) were exposed to these concentration for 96 hours. The fishes were moving slowly as compared to control. No mortalities were found in the control aquaria.  The lethal concentrations LC50 value was determined to be > 100mg/L. Based on the above LC50 value it can be concluded that the target substance 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate is non toxic to fish and thus can not be classified under Aquatic toxic category.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

Data available for short term toxicity to fish endpoint of the substance Disodium 4,4’-(2,4-dihydroxy-5- hydroxymethyl-1,3-phenylene bis-azo) di- (naphthalene-1-sulfonate) are summarized as follows:

In key study, Fish Acute Toxicity test according to OECD Guideline 203 was conducted for Disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate (UERL study report, Report no. 4553 -89 -3/01/2015/FT, Sustainability Support Services (Europe) AB has the letter of access, 2015). The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 1 gm of the test substance in 1 liter of potable water (passed through reverse osmosis system). The nominal concentration selected for the experiment were 6.25 mg/L, 12.5mg/L, 25mg/L, 50mg/L, 100mg/L and Zebra Fish (Danio rerio) were exposed to these concentration for 96 hours. The fishes were moving slowly as compared to control. No mortalities were found in the control aquaria.  The lethal concentrations LC50 value was determined to be > 100mg/L.

While in supporting study, Short term toxicity to fish of Disodium 4,4’-(2,4-dihydroxy-5- hydroxymethyl-1,3-phenylene bis-azo) di- (naphthalene-1-sulfonate) is estimated using QSAR Toolbox version 3.3 (2015). The Lethal concentration (LC50) of the substance Disodium 4,4’-(2,4-dihydroxy-5- hydroxymethyl-1,3-phenylene bis-azo) di- (naphthalene-1-sulfonate) in fish Oncorhynchus mykiss in a 96 hour study is estimated to be 1311.92 mg/L.

Based on the above LC50 values it can be concluded that the target substance 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate is not likely to be toxic to fish and thus cannot be classified under Aquatic toxic category as per CLP regulation.