2-{(E)-[6-({4-chloro-6-[(4-{[2-(sulfooxy)ethyl]sulfonyl}phenyl)amino]-1,3,5-triazin-2-yl}amino)-1-hydroxy-3-sulfonaphthalen-2-yl]diazenyl}naphthalene-1,5-disulfonic acid

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

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:

Experimental data of test chemical

Justification for type of information:

Experimental data of test chemical is from collection of data

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

WoE report is prepared based on short term toxicity to aquatic invertebrate study:
1 and 2nd and 3rd study

GLP compliance:

not specified

Analytical monitoring:

not specified

Vehicle:

yes

Details on test solutions:

1. The solution 100 mg/l was prepared by dissolving red powder in reconstituted water. The solution was kept 30 min in ultrasonic bath.
2. The solution 100 mg/l was prepared by dissolving brown powder in reconstituted water.
3. The solution 100.0 mg/L was prepared by dissolving red powder in reconstituted water. The solution was kept 5 min in ultrasonic bath.

Test organisms (species):

Daphnia magna

Details on test organisms:

1st study: 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

ACCLIMATION - No data available
- Acclimation period:
- Acclimation conditions (same as test or not):
- Type and amount of food:
- Feeding frequency:
- Health during acclimation (any mortality observed):

Test type:

static

Water media type:

freshwater

Total exposure duration:

48 h

Remarks on exposure duration:

1. ± 1 hr

Test temperature:

1. 20±2°C
2 and 3rd: 20±1°C

pH:

1. Test: 7.7 (changed to 7.8 during test)
Control: 7.8 (change to 7.7during test)

2. Test: 7.8 (changed to 7.6 during test)
Control: 7.7 (change to 7.5 during test)

3. Sample : pH = 7.4 changed to pH = 7.8 during the test,
Control: pH = 7.8 changed to pH = 7.7 during the test.

Dissolved oxygen:

1. higher than 7.6 mg/L at the end of test
2. higher than 7.9 mg/L at the end of test
3. Higher than 7.7 mg/L both in the control and the sample.

Nominal and measured concentrations:

1st and 2nd study: 100 mg/l nominal concentration

Details on test conditions:

1, 2nd study:
TEST SYSTEM
- Test vessel: 50 ml glass vessel
- fill volume: 25 ml
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 5

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

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr2O7)

Key result

Duration:

48 h

Dose descriptor:

other: IC8

Effect conc.:

100 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 1st study and 2nd study

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 3rd study

Details on results:

2. 100 mg/l of test chemical was used at which 8% daphnia inhibited after the exposure of 48 hrs.
3. At 100 mg/l only 8% inhibition was observed, thus it can be concluded that the EC50 was >100 mg/l.

Results with reference substance (positive control):

- 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

Validity criteria fulfilled:

not specified

Conclusions:

1. Based on the immobility of daphnia magna due to the test chemical the inhibitory concentration at which only 8% immobility observed was 100 mg/l in a 48 hour study.
2. The effective concentration (EC8) for the test chemical, in Daphnia magna was determined to be 100 mg/L on the basis of mobiity inhibition effects in a 48 hour study.
3. The median effective concentration (EC50) for the test chemical in Daphnia magna was determined to be >100 mg/L on the basis of mobiity inhibition effects in a 48 hour study.
Thus based on the above 3 studies, chemical consider to be non toxic to the aquatic invertebrates and not classified as per the CLP classification criteria.

Executive summary:

Various short term studies available for the test chemical were reviewed to determine the toxic nature of test chemical on the growth and mobility of aquatic invertebrates. The studies are as mentioned below:

An acute immobilisation test was used to test how a range of concentrations of test chemical exerts different degrees of toxic effects on the swimming capability of Daphnia magna under otherwise identical test conditions. The test was performed in close resemblance to OECD guideline 202. The testing aim was to determine a EC50 after 48 hours of exposure to D. magna. Daphnids were exposed to test chemical in 50 ml beakers in a volume of 25 ml of liquid solution containing both the chemical and media as specified in OECD 202. The beakers were placed in a temperature controlled room at 20±1 degrees Celsius. The D. magna (age ≤24) used for the test had been breed at lab. The animals were exposed to medium (i.e. a beaker containing only medium) and the tested chemical during 48 hours (±1 hour). None of the exposed animal’s immobilization were affected by exposure to only medium. The nominal concentrations used were: 100 mg/L (limit test). There were 5 Daphnia per test vessels and 5 replicates per concentration. The pH in test vessels were 7.7-7.8 mg/L. The positive control/reference substance used in the tested showed an expected result and gave an EC50 that corresponded to previous exposures with this chemical in D. magna. The IC50 was defined as a concentration that immobilizes 50% of the exposed D. magna. Eight percent immobilization in D. magna was observed after 48 hours of exposure to 100 mg/L of test chemical. The IC8 was therefore estimated to be 100 mg/L. Based on the IC8, it can be concluded that the chemical was nontoxic and can be consider to be "not classified" as per CLP classification criteria.

First study was supported by the second one. Determination of the inhibition of the mobility of daphnids was carried out with the chemical according to OECD Guideline 202. The limit test was performed at 100 mg/l. Effects on immobilisation were observed for 48 hours. The effective concentration (EC8) for the test chemical,, in Daphnia magna was determined to be 100 mg/L on the basis of mobility inhibition effects in a 48 hour study. 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 criteria.

Similarly determination of the inhibition of the mobility of daphnids was carried out with the test chemical according to OECD Guideline 202. A limit test at sample concentration of 100 mg/L was performed. Effects on immobilisation were observed for 48 hours. At 100 mg/l only 8% inhibition was observed, thus it can be concluded that the EC50 was >100 mg/l. The median effective concentration (EC50) for the test chemical, in Daphnia magna was determined to be >100 mg/L for immobilisation effects. Based on this EC50 value and after comparing with CLP criteria for aquatic classification of the substance it is concluded that the test chemical does not exhibit short term toxicity to aquatic invertebrate (Daphnia Magna).

Thus based on the above 3 studies, chemical consider to be non toxic to the aquatic invertebrates and not classified as per the CLP classification criteria.

Description of key information

Short term toxicity to aquatic invertebrate:

Various short term studies available for the test chemical were reviewed to determine the toxic nature of test chemical on the growth and mobility of aquatic invertebrates. The studies are as mentioned below:

An acute immobilisation test was used to test how a range of concentrations of test chemical exerts different degrees of toxic effects on the swimming capability of Daphnia magna under otherwise identical test conditions. The test was performed in close resemblance to OECD guideline 202. The testing aim was to determine a EC50 after 48 hours of exposure to D. magna. Daphnids were exposed to test chemical in 50 ml beakers in a volume of 25 ml of liquid solution containing both the chemical and media as specified in OECD 202. The beakers were placed in a temperature controlled room at 20±1 degrees Celsius. The D. magna (age ≤24) used for the test had been breed at lab. The animals were exposed to medium (i.e. a beaker containing only medium) and the tested chemical during 48 hours (±1 hour). None of the exposed animal’s immobilization were affected by exposure to only medium. The nominal concentrations used were: 100 mg/L (limit test). There were 5 Daphnia per test vessels and 5 replicates per concentration. The pH in test vessels were 7.7-7.8 mg/L. The positive control/reference substance used in the tested showed an expected result and gave an EC50 that corresponded to previous exposures with this chemical in D. magna. The IC50 was defined as a concentration that immobilizes 50% of the exposed D. magna. Eight percent immobilization in D. magna was observed after 48 hours of exposure to 100 mg/L of test chemical. The IC8 was therefore estimated to be 100 mg/L. Based on the IC8, it can be concluded that the chemical was nontoxic and can be consider to be "not classified" as per CLP classification criteria.

First study was supported by the second one. Determination of the inhibition of the mobility of daphnids was carried out with the chemical according to OECD Guideline 202. The limit test was performed at 100 mg/l. Effects on immobilisation were observed for 48 hours. The effective concentration (EC8) for the test chemical,, in Daphnia magna was determined to be 100 mg/L on the basis of mobility inhibition effects in a 48 hour study. 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 criteria.

Similarly determination of the inhibition of the mobility of daphnids was carried out with the test chemical according to OECD Guideline 202. A limit test at sample concentration of 100 mg/L was performed. Effects on immobilisation were observed for 48 hours. At 100 mg/l only 8% inhibition was observed, thus it can be concluded that the EC50 was >100 mg/l. The median effective concentration (EC50) for the test chemical, in Daphnia magna was determined to be >100 mg/L for immobilisation effects. Based on this EC50 value and after comparing with CLP criteria for aquatic classification of the substance it is concluded that the test chemical does not exhibit short term toxicity to aquatic invertebrate (Daphnia Magna).

Thus based on the above 3 studies, chemical consider to be non toxic to the aquatic invertebrates and not classified as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

Short term toxicity to aquatic invertebrate:

Various short term studies available for the test chemical were reviewed to determine the toxic nature of test chemical on the growth and mobility of aquatic invertebrates. The studies are as mentioned below:

An acute immobilisation test was used to test how a range of concentrations of test chemical exerts different degrees of toxic effects on the swimming capability of Daphnia magna under otherwise identical test conditions. The test was performed in close resemblance to OECD guideline 202. The testing aim was to determine a EC50 after 48 hours of exposure to D. magna. Daphnids were exposed to test chemical in 50 ml beakers in a volume of 25 ml of liquid solution containing both the chemical and media as specified in OECD 202. The beakers were placed in a temperature controlled room at 20±1 degrees Celsius. The D. magna (age ≤24) used for the test had been breed at lab. The animals were exposed to medium (i.e. a beaker containing only medium) and the tested chemical during 48 hours (±1 hour). None of the exposed animal’s immobilization were affected by exposure to only medium. The nominal concentrations used were: 100 mg/L (limit test). There were 5 Daphnia per test vessels and 5 replicates per concentration. The pH in test vessels were 7.7-7.8 mg/L. The positive control/reference substance used in the tested showed an expected result and gave an EC50 that corresponded to previous exposures with this chemical in D. magna. The IC50 was defined as a concentration that immobilizes 50% of the exposed D. magna. Eight percent immobilization in D. magna was observed after 48 hours of exposure to 100 mg/L of test chemical. The IC8 was therefore estimated to be 100 mg/L. Based on the IC8, it can be concluded that the chemical was nontoxic and can be consider to be "not classified" as per CLP classification criteria.

First study was supported by the second one. Determination of the inhibition of the mobility of daphnids was carried out with the chemical according to OECD Guideline 202. The limit test was performed at 100 mg/l. Effects on immobilisation were observed for 48 hours. The effective concentration (EC8) for the test chemical,, in Daphnia magna was determined to be 100 mg/L on the basis of mobility inhibition effects in a 48 hour study. 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 criteria.

Similarly determination of the inhibition of the mobility of daphnids was carried out with the test chemical according to OECD Guideline 202. A limit test at sample concentration of 100 mg/L was performed. Effects on immobilisation were observed for 48 hours. At 100 mg/l only 8% inhibition was observed, thus it can be concluded that the EC50 was >100 mg/l. The median effective concentration (EC50) for the test chemical, in Daphnia magna was determined to be >100 mg/L for immobilisation effects. Based on this EC50 value and after comparing with CLP criteria for aquatic classification of the substance it is concluded that the test chemical does not exhibit short term toxicity to aquatic invertebrate (Daphnia Magna).

Thus based on the above 3 studies, chemical consider to be non toxic to the aquatic invertebrates and not classified as per the CLP classification criteria.