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Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
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:
data from handbook or collection of data
Remarks:
for read across substance
Justification for type of information:
Data for the target chemical is summarized based on the structurally similar read across chemicals
Reason / purpose:
read-across source
Related information:
Composition 1
Reason / purpose:
read-across source
Related information:
Composition 1
Reference:
Composition 0
Qualifier:
according to
Guideline:
other: as mentioned below
Principles of method if other than guideline:
WoE report is based on two short term toxicity study of fish for the test chemical :
1) Short term toxicity of test chemical to aquatic invertebrates was performed according to the OECD guideline in a static system.
2) Short term toxicity study to Artemia Salina was carried out for 24-48 hrs.
Test material information:
Composition 1
Specific details on test material used for the study:
- Name of the test material (IUPAC Name): 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
- Molecular weight: 1035.87  g/mol
- Mol. formula: C28H25N5Na4O20S6
-Smilies:Cc1cc(N=Nc2c(S(=O)(=O)O{-}.[Na]{+})cc3c(ccc(n)c3N=Nc3ccc(S(=O)(=O)CCOS(=O)(=O)O{-}.[Na]{+})cc3S(=O)(=O)O{-}.[Na]{+})c2O)c(OC)cc1S(=O)(=O)CCOS(=O)(=O)O{-}.[Na]{+}
- Substance type: Organic
- Physical state: Solid
Details on sampling:
2) Concentrations: 604.47 and 6044.7 mg/l (10-3 and 10-2 M), respectively.
Sampling method: No data available
Sample storage conditions before analysis: No data available
Test organisms (species):
other: 1)Daphnia magna , 2) Artemia salina
Details on test organisms:
1)TEST ORGANISM
- Common name: Water flea
- Strain: Straus
- Source: Own breeding at University of Chemistry and Technology, Prague
- Age at study initiation (mean and range, SD): The animals used for the test shall be less than 24 h old and should not be first brood progeny
- Feeding during test: No feeding

2) Common name: Brine shrimp
Strain: No data available
Source: A. salina eggs (encysted dried gastrulae) were commercially obtained.
Larvae were obtained by incubating eggs in petri dishes containing muslin-filtered sea water at 30℃ for 24 hours. The larvae were separated from shells, dead larvae and unhatched eggs by their phototactic movements toward a light source.

Test type:
static
Water media type:
freshwater
Total exposure duration:
48 h
Post exposure observation period:
2) After 24 – 48 hrs, larvae surviving were measured by direct count and death rate was calculated.
Test temperature:
1) 20±1°C
2) 30 °C
pH:
1) Test: 7.8 (changed to 7.7 during test)
Control: 7.8 (no change during test)

Dissolved oxygen:
1) higher than 7.8 mg/L at the end of test
Nominal and measured concentrations:
1) 0, 5, 10, 20, 40, 80, 160 mg/l nominal concentration
Details on test conditions:
1) TEST SYSTEM
- Test vessel: 50 ml glass vessel
- fill volume: 25 ml
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Natural water (surface or ground water), reconstituted water or dechlorinated tap water are acceptable as culturing and dilution water if D. magna survives in it for the duration of the culturing, acclimation and testing without showing signs of stress. Waters in the range pH 6 to pH 9, with hardness between 140 mg/l and 275 mg/l (as CaCO3) are recommended.
As an example, the preparation of dilution water meeting the requirements is described below.
Dissolve known quantities of reagents in water. The dilution water prepared shall have a pH of 7.8 ± 0.5, a hardness of (225 ± 50) mg/l (expressed as CaCO3), a molar Ca + Mg ratio close to 4 + 1 and a dissolved oxygen concentration above 7 mg/l.

Prepare the solutions specified below:
- Calcium chloride solution: Dissolve 117.6 g of calcium chloride dihydrate (CaCl2.2H2O) in water (4.2) and make up to 1 l with water (4.2).
- Magnesium sulfate solution: Dissolve 49.3 g of magnesium sulfate heptahydrate (MgSO4.7H2O) in water (4.2) and make up to 1 l with water (4.2).
- Sodium bicarbonate solution: Dissolve 25.9 g of sodium bicarbonate (NaHCO3) in water (4.2) and make up to 1 l with water (4.2).
- Potassium chloride solution: Dissolve 2.3 g of potassium chloride (KCI) in water (4.2) and make up to 1 l with water (4.2).

Mixing
Mix 2.5 ml of each of the four solutions and make up to 1 l with water.
The dilution water shall be aerated until the dissolved oxygen concentration has reached saturation and the pH has stabilized. If necessary, adjust the pH to 7.8 ± 0.5 by adding sodium hydroxide (NaOH) solution or hydrochloric acid (HCI). The dilution water prepared in this way shall not be further aerated before use.

- Sodium hydroxide solution, e.g. [NaOH] : 1 mol/l.
- Hydrochloric acid, e.g. [HCl] : 1 mol/l.

Reference substance:
Dissolve 600 mg of potassium dichromate (K2Cr2O7) in water and make up to 1 l with water (4.2).

OTHER TEST CONDITIONS
- Adjustment of pH: no adjustment done
- Photoperiod: No - Darkness
- Light intensity:

CALCULATION:
EC50 was calculated using non linear regression by the software Prism 4.0

2) Test vessel: Petridish
Type (delete if not applicable): No data available
Material, size, headspace, fill volume: No data available
Aeration: No data available
Type of flow-through (e.g. peristaltic or proportional diluter): No data available
Renewal rate of test solution (frequency/flow rate): No data available
No. of organisms per vessel: 20 – 30 larvae were added to each test vessel.
No. of vessels per concentration (replicates): The test was performed from 5 to 6 six times for each concentration.
No. of vessels per control (replicates):
No. of vessels per vehicle control (replicates): No data available
Biomass loading rate: No data available

TEST MEDIUM / WATER PARAMETERS: No data available

OTHER TEST CONDITIONS: No data available

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): After 24 – 48 hrs, larvae surviving were measured by direct count and death rate was calculated.

TEST CONCENTRATIONS
Spacing factor for test concentrations: No data available
Justification for using less concentrations than requested by guideline: No data available
Range finding study: No data available
Test concentrations: 604.47 and 6044.7 mg/l (10-3 and 10-2 M), respectively.
Results used to determine the conditions for the definitive study: No data available
Reference substance (positive control):
yes
Remarks:
Potassium dichromate (K2Cr2O7)
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 900 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
24 h
Dose descriptor:
LC0
Effect conc.:
604.477 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: No mortality of test organism was observed and death rate was found to be 0%.
Results with reference substance (positive control):
1) - Results with reference substance valid
- EC50: 0.79 mg/L (24 hours)
Reported statistics and error estimates:
1) EC50 was calculated using non linear regression by the software Prism 4.0
Conclusions:
The test chemical 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 is not likely to be toxic to aquatic invertebrate atleast in the concentration range of 600-900 mg/l
Executive summary:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the short term toxicity of aquatic invertebrates of the test chemical 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 (503155 -49 -5).The studies are as mentioned below:

1) Aim of the study was to assess the effect of chemical tetrasodium on the mobility of daphnia magna. Test was conducted according to OECD Guideline 202. The stock solution 100 mg/l was prepared by dissolving red powder in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water. The test substance was tested at the 0, 5, 10, 20, 40, 80, 160 mg/l nominal concentration. Potassium dichromate (K2Cr2O7) were used as a reference positive control.

Effects on immobilisation were observed for 48 hours by using non linear regression by the software Prism 4.0. After the exposure of chemical for 48 hrs 50 % immobility was observed. Based on the immobility of daphnia magna, the median effective concentration (EC50) for the test substance, in Daphnia magna was determined to be > 900 mg/L for immobilisation effects.

This value indicates that the substance is likely to be non-hazardous to aquatic invertebrates and cannot be classified as toxic as per the CLP classification criteria.

 

2) The toxic effects of test chemical were studied on Artemia salinalarvae. Artemia salina (A. salina eggs) a crustacean, commonly known as brine shrimp eggs, are commercially available, and are easily cultured in the laboratory because they are resistant to environmental stresses. Active larvae can be obtained within 1 to 2 days and no live culture is required for a few days thereafter.

 

A. salina eggs (encysted dried gastrulae) were commercially obtained, and were stored at -200°C. Eggs used in experiments were washed and stored at room temperature in a desiccators over anhydrous granular CaCl2. Larvae were obtained by incubating eggs in petri dishes containing muslin-filtered sea water at 30°C for 24 hours. The larvae were separated from shells, dead larvae and unhatched eggs by their phototactic movements towards a light source. Amaranth at concentrations of 6044.7mg/l and 604.47 mg/l were placed in a petri dish, and sea water containing 20 to 30 larvae was added. After this was incubated at 30°C for 24 hours and 48 hours, larvae surviving were measured by direct count. The same method was tested from 5 to 6 times for each concentration, and the death rate was calculated. Death was assumed to have occurred when there was no movement. The death rate was defined as the average of the percentage of deaths observed for 24 hours and 48 hours.100% death rate was noted after 48 hours when 6044.7 mg/l of test material was exposed to the test organism and 0% death rate after 24 hours in case of exposure to 604.47 mg/l of test chemical.

Thus, based on the above summarised studies, 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 and it’s structurally and functionally similar read across substance, it can be concluded that effect concetration value is greater than 900 mg/L. Thus, comparing this value with the criteria of CLP regulation,2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2 - cannot be classified for short term toxicity of aquatic invertebrate .Hence,based on the data available for the structurally and functionally similar read across, test chemical 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 is not likely to be toxic atleast in the concentration range of 600 -900 mg/L .(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt .

Description of key information

Short term toxicity to aquatic invertebrate:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the short term toxicity of aquatic invertebrates of the test chemical 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 (503155 -49 -5).The studies are as mentioned below:

1)Aim of the study was to assess the effect of chemical tetrasodium on the mobility of daphnia magna. Test was conducted according to OECD Guideline 202. The stock solution 100 mg/l was prepared by dissolving red powder in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water. The test substance was tested at the 0, 5, 10, 20, 40, 80, 160 mg/l nominal concentration. Potassium dichromate (K2Cr2O7) were used as a reference positive control.

Effects on immobilisation were observed for 48 hours by using non linear regression by the software Prism 4.0. After the exposure of chemical for 48 hrs 50 % immobility was observed. Based on the immobility of daphnia magna, the median effective concentration (EC50) for the test substance, in Daphnia magna was determined to be > 900 mg/L for immobilisation effects.

This value indicates that the substance is likely to be non-hazardous to aquatic invertebrates and cannot be classified as toxic as per the CLP classification criteria.

 

2)The toxic effects of test chemical were studied on Artemia salinalarvae. Artemia salina (A. salina eggs) a crustacean, commonly known as brine shrimp eggs, are commercially available, and are easily cultured in the laboratory because they are resistant to environmental stresses. Active larvae can be obtained within 1 to 2 days and no live culture is required for a few days thereafter.

 

A. salina eggs (encysted dried gastrulae) were commercially obtained, and were stored at -200°C. Eggs used in experiments were washed and stored at room temperature in a desiccators over anhydrous granular CaCl2. Larvae were obtained by incubating eggs in petri dishes containing muslin-filtered sea water at 30°C for 24 hours. The larvae were separated from shells, dead larvae and unhatched eggs by their phototactic movements towards a light source. Amaranth at concentrations of 6044.7mg/l and 604.47 mg/l were placed in a petri dish, and sea water containing 20 to 30 larvae was added. After this was incubated at 30°C for 24 hours and 48 hours, larvae surviving were measured by direct count. The same method was tested from 5 to 6 times for each concentration, and the death rate was calculated. Death was assumed to have occurred when there was no movement. The death rate was defined as the average of the percentage of deaths observed for 24 hours and 48 hours.100% death rate was noted after 48 hours when 6044.7 mg/l of test material was exposed to the test organism and 0% death rate after 24 hours in case of exposure to 604.47 mg/l of test chemical.

Thus, based on the above summarised studies, 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 and it’s structurally and functionally similar read across substance, it can be concluded that effect concetration value is greater than 900 mg/L. Thus, comparing this value with the criteria of CLP regulation,2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2 -cannot be classified for short term toxicity of aquatic invertebrate .Hence,based on the data available for the structurally and functionally similar

read across, test chemical 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 is not likely to be toxic atleast in the concentration range of 600 -900 mg/L .(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt .

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
900 mg/L

Additional information

Short term toxicity to aquatic invertebrate:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the short term toxicity of aquatic invertebrates of the test chemical 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 (503155 -49 -5).The studies are as mentioned below:

1)Aim of the study was to assess the effect of chemical tetrasodium on the mobility of daphnia magna. Test was conducted according to OECD Guideline 202. The stock solution 100 mg/l was prepared by dissolving red powder in reconstituted water. Test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water. The test substance was tested at the 0, 5, 10, 20, 40, 80, 160 mg/l nominal concentration. Potassium dichromate (K2Cr2O7) were used as a reference positive control.

Effects on immobilisation were observed for 48 hours by using non linear regression by the software Prism 4.0. After the exposure of chemical for 48 hrs 50 % immobility was observed. Based on the immobility of daphnia magna, the median effective concentration (EC50) for the test substance, in Daphnia magna was determined to be > 900 mg/L for immobilisation effects.

This value indicates that the substance is likely to be non-hazardous to aquatic invertebrates and cannot be classified as toxic as per the CLP classification criteria.

 

2)The toxic effects of test chemical were studied on Artemia salinalarvae. Artemia salina (A. salina eggs) a crustacean, commonly known as brine shrimp eggs, are commercially available, and are easily cultured in the laboratory because they are resistant to environmental stresses. Active larvae can be obtained within 1 to 2 days and no live culture is required for a few days thereafter.

 

A. salina eggs (encysted dried gastrulae) were commercially obtained, and were stored at -200°C. Eggs used in experiments were washed and stored at room temperature in a desiccators over anhydrous granular CaCl2. Larvae were obtained by incubating eggs in petri dishes containing muslin-filtered sea water at 30°C for 24 hours. The larvae were separated from shells, dead larvae and unhatched eggs by their phototactic movements towards a light source. Amaranth at concentrations of 6044.7mg/l and 604.47 mg/l were placed in a petri dish, and sea water containing 20 to 30 larvae was added. After this was incubated at 30°C for 24 hours and 48 hours, larvae surviving were measured by direct count. The same method was tested from 5 to 6 times for each concentration, and the death rate was calculated. Death was assumed to have occurred when there was no movement. The death rate was defined as the average of the percentage of deaths observed for 24 hours and 48 hours.100% death rate was noted after 48 hours when 6044.7 mg/l of test material was exposed to the test organism and 0% death rate after 24 hours in case of exposure to 604.47 mg/l of test chemical.

Thus, based on the above summarised studies, 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 and it’s structurally and functionally similar read across substance, it can be concluded that effect concetration value is greater than 900 mg/L. Thus, comparing this value with the criteria of CLP regulation,2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-3-[[2-methoxy-5-methyl-4-[[2 -cannot be classified for short term toxicity of aquatic invertebrate .Hence,based on the data available for the structurally and functionally similar

read across, test chemical 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 is not likely to be toxic atleast in the concentration range of 600 -900 mg/L .(sulfooxy)ethyl]sulfonyl] phenyl]azo]-8-[[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, tetrasodium salt .