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 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

Principles of method if other than guideline:

WoE report is based on two short term toxicity to aquatic invertebrate 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 stock solution 150 mg/l was prepared by dissolving test chemical in reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample in reconstituted test water.
WoE 3: The solution 100 g/l was prepared by dissolving test chemical in reconstituted water. The solution was kept 5 min in ultrasonic bath.

Test organisms (species):

Daphnia magna

Details on test organisms:

WoE 2 and WoE 3:
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

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

± 1 hr

Test temperature:

WoE 2 and WoE 3: 20±1°C

pH:

WoE 2:
without adjustment
sample at concentration 150 mg/l: pH = 7.8 changed to pH = 7.7 during the test
control 1: pH = 7.7 changed to pH = 7.5 during the test
control 2: pH = 7.8 changed to pH = 7.7 during the test

WoE 3:
without adjustment
sample: pH = 7.6 changed to pH = 7.8 during the test
control: pH = 7.8 changed to pH = 7.7 during the test

Dissolved oxygen:

WoE 2: higher than 7.9 mg/L at the end of test both in the control and the sample
WoE 3: higher than 7.5 mg/L at the end of test both in the control and the sample

Nominal and measured concentrations:

WoE 2: Nominal test chemical concentration used for the study were 0,10, 45, 67.5, 100 and 150 mg/l, respectively.
WoE 3: Nominal test chemical concentration used for the study was 100 mg/l.

Details on test conditions:

WoE 2:
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: Darkness
- Light intensity:

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

Reference substance (positive control):

yes

Remarks:

WoE 2 and WoE 3: Potassium dichromate (K2Cr2O7)

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

203.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: WoE 2: 95 % C. I. - 140.9 to 293.2 mg/l

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 3

Results with reference substance (positive control):

WoE 2:
- Results with reference substance valid
- EC50: 0.79 mg/L (24 hours)

WoE 3:
- Results with reference substance valid
- EC50: 0.79 mg/L (24 hours)

Reported statistics and error estimates:

WoE 2: EC50 was calculated using non linear regression by the software Prism 4.0.
WoE 3: no data available

WoE 2:

In control solution containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test.

Result of the definitive test:

Sample no.	Sample Information	Conc. (mg/l)	I%	48 hr EC50
				mg/l	95% C. I. (mg/l)
Control Control	Reconstituted water	0 0	0 0
Test chemical	Test chemical	10 45 67.5 100.0 150.0	0 15 25 32 40	203.2	140.9 - 293.2

WoE 3:

In control solution containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test.

Result of a limit test at sample concentration 100 mg/l:

Sample information	Control (number of mobile organisms)	Sample (number of mobile organisms)	I (%)
Test chemical	25	25	0

 

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 48 hr EC50 value of the test chemical on test organism can be expected to be in the range of >100 to 203.2 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 aquatic invertebrates. The studies are as mentioned below:

An acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on aquatic invertebrates. The test was performed in accordance to OECD guideline No. 202 “Daphnia sp.,Acute Immobilization Test”. Daphnia magna was used as a test organism for the study. The stock solution 150 mg/l was prepared by dissolving test chemical in reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample in reconstituted test water. Nominal test chemical concentrations used for the study were 0,10, 45, 67.5, 100 and 150 mg/l, respectively. Study was performed using 5 organisms per vessel/replicates in a static system. Daphnids were exposed to test chemical in 50 ml glass vessel in a volume of 25 ml of liquid solution containing both the chemical and media. Control solution vessel containing reconstituted water without the test chemical was also setup during the study. The beakers were placed in a room at a temperature of 20±1°C. With the test substance one positive control Potassium dichromate (K2Cr2O7) was run simultaneously. EC50 was calculated using non-linear regression by the software Prism 4. EC50 value of the reference substance was determined to be 0.79 mg/l. In the control vessel containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test. On the basis of the mobility of the test organism Daphnia magna due to the exposure of test chemical, the 48hr median effect concentration (EC50) value was determined to be 203.2 mg/l (95% C. I. - 140.9 to 293.2 mg/l).

Another acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on aquatic invertebrates. The test was performed in accordance to OECD guideline No. 202 “Daphnia sp.,Acute Immobilization Test”. Daphnia magna was used as a test organism for the study. The solution 100 mg/l was prepared by dissolving test chemical in reconstituted water. The solution was kept 5 min in ultrasonic bath. Thus, limit test was performed using a nominal test conc. of 100 mg/l. Study was performed using 5 organisms per vessel/replicates in a static system. Daphnids were exposed to test chemical in 50 ml glass vessel in a volume of 25 ml of liquid solution containing both the chemical and media. Control solution vessel containing reconstituted water without the test chemical was also run simulatenously during the study. The beakers were placed in a room at a temperature of 20±1°C. With the test substance one positive control Potassium dichromate (K2Cr2O7) was run simultaneously. In the control vessel containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test. On the basis of the mobility of the test organism Daphnia magna due to the exposure of test chemical, the 48hr median effect concentration (EC50) value was determined to be >100 mg/l (with percent immobility as 0 at 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 48 hr EC50 value of the test chemical on test organism can be expected to be in the range of >100 to 203.2 mg/l. Thus, test chemical was considered as non-toxic to aquatic invertebrates 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 48 hr EC50 value of the test chemical on test organism can be expected to be in the range of >100 to 203.2 mg/l.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

203.2 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 aquatic invertebrates. The studies are as mentioned below:

 

An acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on aquatic invertebrates. The test was performed in accordance to OECD guideline No. 202 “Daphnia sp.,Acute Immobilization Test”. Daphnia magna was used as a test organism for the study. The stock solution 150 mg/l was prepared by dissolving test chemical in reconstituted water. Test solutions of required concentrations were prepared by mixing the stock solution of the test sample in reconstituted test water. Nominal test chemical concentrations used for the study were 0,10, 45, 67.5, 100 and 150 mg/l, respectively. Study was performed using 5 organisms per vessel/replicates in a static system. Daphnids were exposed to test chemical in 50 ml glass vessel in a volume of 25 ml of liquid solution containing both the chemical and media. Control solution vessel containing reconstituted water without the test chemical was also setup during the study. The beakers were placed in a room at a temperature of 20±1°C. With the test substance one positive control Potassium dichromate (K2Cr2O7) was run simultaneously. EC50 was calculated using non-linear regression by the software Prism 4. EC50 value of the reference substance was determined to be 0.79 mg/l. In the control vessel containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test. On the basis of the mobility of the test organism Daphnia magna due to the exposure of test chemical, the 48hr median effect concentration (EC50) value was determined to be 203.2 mg/l (95% C. I. - 140.9 to 293.2 mg/l).

 

Another acute immobilisation test was conducted for 48 hrs for assessing the effect of test chemical on aquatic invertebrates. The test was performed in accordance to OECD guideline No. 202 “Daphnia sp.,Acute Immobilization Test”. Daphnia magna was used as a test organism for the study. The solution 100 mg/l was prepared by dissolving test chemical in reconstituted water. The solution was kept 5 min in ultrasonic bath. Thus, limit test was performed using a nominal test conc. of 100 mg/l. Study was performed using 5 organisms per vessel/replicates in a static system. Daphnids were exposed to test chemical in 50 ml glass vessel in a volume of 25 ml of liquid solution containing both the chemical and media. Control solution vessel containing reconstituted water without the test chemical was also run simulatenously during the study. The beakers were placed in a room at a temperature of 20±1°C. With the test substance one positive control Potassium dichromate (K2Cr2O7) was run simultaneously. In the control vessel containing reconstituted water without the test chemical, no daphnids were immobilized at the end of the test. On the basis of the mobility of the test organism Daphnia magna due to the exposure of test chemical, the 48hr median effect concentration (EC50) value was determined to be >100 mg/l (with percent immobility as 0 at 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 48 hr EC50 value of the test chemical on test organism can be expected to be in the range of >100 to 203.2 mg/l. Thus, test chemical was considered as non-toxic to aquatic invertebrates and hence, considered to be 'not classified' as per the CLP classification criteria.