Calcium 4-[(4-chloro-3-sulphonatophenyl)azo]-3-hydroxy-2-naphthoate

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:

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 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 principle below

Principles of method if other than guideline:

WoE report is prepared based on short term toxicity to aquatic invertebrate studies namely WoE-2, WoE- 3 and WoE-4

GLP compliance:

no

Analytical monitoring:

not specified

Vehicle:

yes

Details on test solutions:

WoE-2:
- Chemical name of vehicle : Control of DMSO: HCO-40 is 9:1 (100 mg/l)
- Controls: laboratory water control.

WoE-3 :PREPARATION AND APPLICATION OF TEST SOLUTION
The solution 100.0 mg/L was prepared by dissolving test chemical in reconstituted water. The solution
was kept 5 min in ultrasonic bath.
- Controls: Reconstituted water as specified in OECD 202, used as a control and for sample dilution.

WoE-4: The stock solution 100 mg/L was prepared by dissolving dark red powder in reconstituted water.

Test organisms (species):

Daphnia magna

Details on test organisms:

WoE-2:TEST ORGANISM- Common name: Water flea

WoE-3:TEST ORGANISM
- Common name: Water flea
- Strain/clone: Straus
- Source: Own breeding at University of Chemistry and Technology, Prague
- Age: Young daphnids, aged less than 24 hours used at the start of the test and they should not be
first brood progeny.
- Feeding during test : Without feeding


WoE-4: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

Total exposure duration:

48 h

Test temperature:

WoE-3: 20 °C ± 1 °C

WoE-4: 20±1°C

pH:

WoE-3:Sample : pH = 7.6 changed to pH = 7.8 during the test,
Control: pH = 7.8 changed to pH = 7.7 during the test.

WoE-4:sample: pH = 7.8 changed to pH = 7.9 during the test,
control: pH = 8.0 changed to pH = 7.9 during the test

Dissolved oxygen:

WoE-3:Higher than 7.5 mg/L both in the control and the sample.

WoE-4: higher than 8.3 mg/L at the end of test

Nominal and measured concentrations:

WoE-2: 5 nominal concentrations (90-940 mg/l)

WoE-4: 100 mg/L

Details on test conditions:

WoE-2:
- Type: Closed
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): 4

WoE-3:TEST SYSTEM
- Test vessel: 50 ml glass vessel
- Material, size, headspace, fill volume: 25 ml
- Aeration: no data
- No. of organisms per vessel: 5 pieces
- 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.
OTHER TEST CONDITIONS
- Adjustment of pH: Without adjustment
- Photoperiod: No - Darkness
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : EC50 was
calculated.
Reference substance (positive control)
yes Potassium dichromate (K2Cr2O7)

WoE-4: 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
Reference substance (positive control)

Reference substance (positive control):

not specified

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

280 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

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

Key result

Duration:

48 h

Dose descriptor:

other: I %

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: other: at concentration 100 mg/L,8.0 % inhibition of mobility of organisms was observed.

Results with reference substance (positive control):

WoE-3: Results with reference substance valid
- 24h EC50 = 0.79 mg/L K2Cr2O7

WoE-4: - Results with reference substance valid
- EC50: 0.73 mg/L (24 hours)

Reported statistics and error estimates:

WoE-4: EC50 was calculated using non linear regression by the software Prism 4.0

Validity criteria fulfilled:

not specified

Conclusions:

By applying weight of evidence approach the EC50 value of test chemical was determined to be >100 mg/L after exposure of test chemical for 48 hrs. The EC50 value indicates that the substance is likely to be nonhazardous to aquatic invertebrates and cannot be classified as per the CLP classification criteria

Executive summary:

Data available for the structurally and functionally read across chemicals has been reviewed to determine the short term toxicity to aquatic invertebrates. The studies are as mentioned below:

The first study reviewed from the secondary source and this study was performed to determine the effect of test chemical on Daphnia magna according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). The vehicle used in the study was DMSO and HCO-40 at ratio of DMSO: HCO-40 is 9:1 (100 mg/l), and laboratory water was used as negative control.

The test was performed in static system. The 5 nominal concentrations of test chemical were used and they are in range of 90-940 mg/l. The static procedure was used in this study and nominal concentrations of test chemical used were exposed to Daphnia magna for 24 hrs.

The effect concentration of test chemical at which 50 % of Daphnia magna were immobilized was determined to be 280 mg/l.

Another study was performed to determine the effect of test chemical on Daphnia magna according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). The test chemical solution 100.0 mg/L was prepared by dissolving test chemical in reconstituted water. The solution was kept 5 min in ultrasonic bath. Limit test was performed in this study at concentration 100 mg/L. The positive control used was Potassium dichromate (K2Cr2O7). Static system was used in this study and test chemical was exposed to Daphnia magna for 48 hrs. The median effective concentration (EC50) of test chemical, in Daphnia magna was determined to be >100 mg/L. Based on this EC50 value test chemical is considered to be non toxic to Daphnia Magna and cannot be classified as per CLP regulation.

Aim of this next study was to assess the short term toxicity of test chemical to aquatic invertebrate daphnia magna. Study was performed according to the OECD Guideline 202 (Daphnia sp. The test chemical solution of concentration 100 mg/L was prepared by dissolving test chemical in reconstituted water. Limit test was performed in this study at concentration 100 mg/L .Effects on immobilization were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, inhibition percentage (I%) was calculated using nonlinear regression by the software Prism 4.0. The inhibition percentage (I%) for the test substance in Daphnia magna was determined to be 8.0 % at 100 mg/L on the basis of mobility inhibition effects . Based on the percentage (I%) value, substance is considered to be non-hazardous to aquatic invertebrate and cannot be classified as per the CLP classification criteria.

By considering results of both the studies mentioned above and applying weight of evidence approach the EC50 value of test chemical was determined to be >100 mg/L after exposure of test chemical for 48 hrs. The EC50 value indicates that the substance is likely to be nonhazardous to aquatic invertebrates and cannot be classified as per the CLP classification criteria

Description of key information

By applying weight of evidence approach the EC50 value of test chemical was determined to be >100 mg/L after exposure of test chemical for 48 hrs. The EC50 value indicates that the substance is likely to be nonhazardous to aquatic invertebrates and cannot be 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

Data available for the structurally and functionally read across chemicals has been reviewed to determine the short term toxicity to aquatic invertebrates. The studies are as mentioned below:

The first study reviewed from the secondary source and this study was performed to determine the effect of test chemical on Daphnia magna according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). The vehicle used in the study was DMSO and HCO-40 at ratio of DMSO: HCO-40 is 9:1 (100 mg/l), and laboratory water was used as negative control.

The test was performed in static system. The 5 nominal concentrations of test chemical were used and they are in range of 90-940 mg/l. The static procedure was used in this study and nominal concentrations of test chemical used were exposed to Daphnia magna for 24 hrs.

The effect concentration of test chemical at which 50 % of Daphnia magna were immobilized was determined to be 280 mg/l with 95% confidence level of 150-490 mg/L.

Another study was performed to determine the effect of test chemical on Daphnia magna according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). The test chemical solution 100.0 mg/L was prepared by dissolving test chemical in reconstituted water. The solution was kept 5 min in ultrasonic bath. Limit test was performed in this study at concentration 100 mg/L. The positive control used was Potassium dichromate (K2Cr2O7). Static system was used in this study and test chemical was exposed to Daphnia magna for 48 hrs. The median effective concentration (EC50) of test chemical, in Daphnia magna was determined to be >100 mg/L. Based on this EC50 value test chemical is considered to be non toxic to Daphnia Magna and cannot be classified as per CLP regulation.

Aim of this next study was to assess the short term toxicity of test chemical to aquatic invertebrate daphnia magna. Study was performed according to the OECD Guideline 202 (Daphnia sp. The test chemical solution of concentration 100 mg/L was prepared by dissolving test chemical in reconstituted water. Limit test was performed in this study at concentration 100 mg/L .Effects on immobilization were observed for 48 hours. With the test substance one positive control Potassium dichromate (K2Cr2O7) was also run simultaneously. After the exposure of chemical, inhibition percentage (I%) was calculated using nonlinear regression by the software Prism 4.0. The inhibition percentage (I%) for the test substance in Daphnia magna was determined to be 8.0 % at 100 mg/L on the basis of mobility inhibition effects by considering this EC50 value of test chemical was determined to be >100 mg/L

. Based on the percentage (I%) value, substance is considered to be non-hazardous to aquatic invertebrate and cannot be classified as per the CLP classification criteria.

By considering results of both the studies mentioned above and applying weight of evidence approach the EC50 value of test chemical was determined to be >100 mg/L after exposure of test chemical for 48 hrs. The EC50 value indicates that the substance is likely to be nonhazardous to aquatic invertebrates and cannot be classified as per the CLP classification criteria