5-((4-chloro-6-((8-hydroxy-3,6-disulfo-7-((2-sulfo-4-((2(sulfooxy)ethyl)sulfonyl)phenyl)diazenyl)naphthalen-1-yl)amino)-1,3,5-triazin-2-yl)amino)-4-hydroxy-3-((Z)-(3-sulfophenyl)diazenyl)naphthalene-2,7-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 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

GLP compliance:

not specified

Analytical monitoring:

not specified

Details on sampling:

WoE 2 and WoE 3:
- Concentrations: Test chemical concentration used for the study was 100 mg/l.
- Sampling method: Test chemical solutions were prepared by directly dissolving the test chemical in specific test medium (ISO 6341 Standard Freshwater).

Vehicle:

not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

WoE 2 and WoE 3:
TEST ORGANISM
- Common name: Water flea
- Source: Test organism was taken from Daphtoxkit F™ magna, a commercial
kit that provides daphnids in the form of dormant eggs (ephippia) that need to be incubate 3 days before the use to obtain neonates used for the test.
- Other: Neonates less than 24 h old (25 for each condition) were fed with lyophilized spirulina as dietary supplement for 2 h prior to the test and then moved to the exposure chamber.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

WoE 2 and WoE 3: 20 ± 1°C

Details on test conditions:

WoE 2 and WoE 3:
TEST SYSTEM
- No. of organisms per vessel: 25 organisms

TEST MEDIUM / WATER PARAMETERS: ISO 6341 Standard Freshwater was used as a test medium for the study. Media were aerated for at least 1 h prior to use in order to ensure that the dissolved oxygen concentration has reached saturation

OTHER TEST CONDITIONS
- Light intensity: no illumination of light was provided to the test vessel.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilization of daphnids was recorded daily.

Reference substance (positive control):

yes

Remarks:

WoE 2 and WoE 3: Potassium dichromate (0.32 – 3.2 mg/L) was used as a reference substance during the study.

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: WoE 2 and WoE 3

Reported statistics and error estimates:

WoE 2 and WoE 3: 48 h LC50 were derived by non-linear regression analysis. All statistical analyses were done using Prism5 (GraphPad Software, Inc.). One-way ANOVA was used for statistical comparisons with Bonferroni's, Dunnett's and Tukey's post hoc tests for multiple comparisons. Significance was set at p<0.05. REGTOX macro Excel™ for dose-response was used to obtain EC50 values by a non-linear regression analysis with Hill's model.

WoE 2 and WoE 3: Dissolved oxygen concentration was measured with HI9142 Portable Dissolved Oxygen Meter (Hanna Instruments srl) in all test chambers at the end of the test and was higher than 3 mg/l, according to the OECD 202 guidelines.

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 and by evaluating the effect of test chemical
on test organism, the 48 hr EC50 value can be expected to be >100 mg/l.

Executive summary:

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

An acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical.The test was performed following the principles of the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test).The study was performed under static conditions using Daphnia magna (Water flea) as a test organism. Test organism was taken from Daphtoxkit F™ magna, a commercial kit that provides daphnids in the form of dormant eggs (ephippia) that need to be incubate 3 days before the use to obtain neonates used for the test. Neonates less than 24 h old (25 for each condition) were fed with lyophilized spirulina as dietary supplement for 2 h prior to the test and then moved to the exposure chamber. Test chemical solutions were prepared by directly dissolving the test chemical in specific test medium (ISO 6341 Standard Freshwater). Test chemical concentration used for the study was 100 mg/l. Thus, a limit test was performed using 100 mg/l test chemical concentration. Selected bottled water was used only in the daphnid assays and satisfied the acceptability criteria for holding and dilution water described by the OECD 202 guideline. ISO 6341 Standard Freshwater was used as a test medium for the study. Media were aerated for at least 1 h prior to use in order to ensure that the dissolved oxygen concentration has reached saturation. Test daphnids (25 test organisms) were exposed with the test chemical at 20±1°C for a period of 48 hrs. No illumination of light was provided to the test vessel during the study. No significant changes in pH were recorded in the test solutions. Untreated ISO formulation freshwater was used as negative control. Potassium dichromate (0.32 – 3.2 mg/L) was used as a reference substance during the study. 48 h LC50 were derived by non-linear regression analysis. All statistical analyses were done using Prism5 (GraphPad Software, Inc.). One-way ANOVA was used for statistical comparisons with Bonferroni's, Dunnett's and Tukey's post hoc tests for multiple comparisons. Significance was set at p<0.05. REGTOX macro Excel™ for dose-response was used to obtain EC50 values by a non-linear regression analysis with Hill's model. Dissolved oxygen concentration was measured with HI9142 Portable Dissolved Oxygen Meter (Hanna Instruments srl) in all test chambers at the end of the test and was higher than 3 mg/l, according to the OECD 202 guidelines.On the basis of the toxic effect of the test chemical on mobility of the test organism Daphnia magna,the 48 hr median lethal concentration (LC50) value was determined to be >100 mg/l (nominal conc.). Thus, test chemical was consideredas non-toxic to aquatic invertebrates at environmental relevant concentrations andhence, considered to be ‘not classified’ as per the CLP classification criteria.

Another short term toxicity to aquatic invertebrate study was conducted for 48 hrs for assessing the effect of test chemical.The test was performed following the principles of the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test).The study was performed under static conditions using Daphnia magna (Water flea) as a test organism. Test was conducted using the same test procedure and test conditions as mentioned in above study. On the basis of the toxic effect of the test chemical on mobility of the test organism Daphnia magna, the 48 hr median lethal concentration (LC50) value was determined to be >100 mg/l (nominal conc.). Thus, test chemical was consideredas non-toxic to aquatic invertebrates at environmental relevant concentrations andhence, considered to be ‘not classified’ as per the CLP classification criteria.

On the basis of the experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach and by evaluating the effect of test chemical on test organism, the 48 hr EC50 value can be expected to be >100 mg/l.

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 and by evaluating the effect of test chemical on test organism, the 48 hr EC50 value can be expected to be >100 mg/l.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

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

 

An acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical. The test was performed following the principles of the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test).The study was performed under static conditions using Daphnia magna (Water flea) as a test organism. Test organism was taken from Daphtoxkit F™ magna, a commercial kit that provides daphnids in the form of dormant eggs (ephippia) that need to be incubate 3 days before the use to obtain neonates used for the test. Neonates less than 24 h old (25 for each condition) were fed with lyophilized spirulina as dietary supplement for 2 h prior to the test and then moved to the exposure chamber. Test chemical solutions were prepared by directly dissolving the test chemical in specific test medium (ISO 6341 Standard Freshwater). Test chemical concentration used for the study was 100 mg/l. Thus, a limit test was performed using 100 mg/l test chemical concentration. Selected bottled water was used only in the daphnid assays and satisfied the acceptability criteria for holding and dilution water described by the OECD 202 guideline. ISO 6341 Standard Freshwater was used as a test medium for the study. Media were aerated for at least 1 h prior to use in order to ensure that the dissolved oxygen concentration has reached saturation. Test daphnids (25 test organisms) were exposed with the test chemical at 20±1°C for a period of 48 hrs. No illumination of light was provided to the test vessel during the study. No significant changes in pH were recorded in the test solutions. Untreated ISO formulation freshwater was used as negative control. Potassium dichromate (0.32 – 3.2 mg/L) was used as a reference substance during the study. 48 h LC50 were derived by non-linear regression analysis. All statistical analyses were done using Prism5 (GraphPad Software, Inc.). One-way ANOVA was used for statistical comparisons with Bonferroni's, Dunnett's and Tukey's post hoc tests for multiple comparisons. Significance was set at p<0.05. REGTOX macro Excel™ for dose-response was used to obtain EC50 values by a non-linear regression analysis with Hill's model. Dissolved oxygen concentration was measured with HI9142 Portable Dissolved Oxygen Meter (Hanna Instruments srl) in all test chambers at the end of the test and was higher than 3 mg/l, according to the OECD 202 guidelines. On the basis of the toxic effect of the test chemical on mobility of the test organism Daphnia magna, the 48 hr median lethal concentration (LC50) value was determined to be >100 mg/l (nominal conc.). Thus, test chemical was consideredas non-toxic to aquatic invertebrates at environmental relevant concentrations and hence, considered to be ‘not classified’ as per the CLP classification criteria.

 

Another short term toxicity to aquatic invertebrate study was conducted for 48 hrs for assessing the effect of test chemical. The test was performed following the principles of the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test).The study was performed under static conditions using Daphnia magna (Water flea) as a test organism. Test was conducted using the same test procedure and test conditions as mentioned in above study. On the basis of the toxic effect of the test chemical on mobility of the test organism Daphnia magna, the 48 hr median lethal concentration (LC50) value was determined to be >100 mg/l (nominal conc.). Thus, test chemical was considered as non-toxic to aquatic invertebrates at environmental relevant concentrations and hence, considered to be ‘not classified’ as per the CLP classification criteria.

 

On the basis of the experimental studies of the structurally and functionally similar read across chemical and applying the weight of evidence approach and by evaluating the effect of test chemical on test organism, the 48 hr EC50 value can be expected to be >100 mg/l.