Dihydrogen (ethyl)[4-[[4-[ethyl(3-sulphonatobenzyl)amino]phenyl](2-sulphonatophenyl)methylene]cyclohexa-2,5-dien-1-ylidene](3-sulphonatobenzyl)ammonium, aluminium salt

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

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 four toxicity to microorganism studies as-
WoE 2 and WoE 3.

GLP compliance:

no

Analytical monitoring:

no

Remarks:

WoE 2: no and WoE 3: not specified

Vehicle:

no

Remarks:

WoE 2: no and WoE 3: not specified

Details on test solutions:

WoE 2: The solution 200 mg/l was prepared by dissolving test chemical in reconstituted water.
WoE 3: no data available

Test organisms (species):

Daphnia magna

Details on test organisms:

WoE 2:
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

WoE 3:
TEST ORGANISM
- Common name: Water flea

Test type:

static

Water media type:

freshwater

Total exposure duration:

48 h

Test temperature:

WoE 2: 20±1°C
WoE 3: no data available

pH:

WoE 2:
without adjustment
sample: pH = 7.5 changed to pH = 7.8 during the test
control: pH = 7.8 did not change during the test

WoE 3: no data available

Dissolved oxygen:

WoE 2: higher than 8.9 mg/L at the end of test both in the control and the sample
WoE 3: no data available

Nominal and measured concentrations:

WoE 2: Nominal test chemical concentration used for the study was 200 mg/l.
WoE 3: no data available

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: No - Darkness
- Light intensity:

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

WoE 3: no data available

Reference substance (positive control):

yes

Remarks:

WoE 2: Potassium dichromate (K2Cr2O7) and WoE 3: no data available

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 200 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: WoE 2

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: WoE 3

Results with reference substance (positive control):

WoE 2:
- Results with reference substance valid
- EC50: 0.76 mg/L (24 hours)

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 a limit test at sample concentration 200 mg/l:

Sample no.	Sample information	Control (number of mobile organisms)	Sample (number of mobile organisms)	I (%)
13244	Test chemical	25	24	4.0

WoE 3: no data available 

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/LC50 value can be expected to be in the range 200 to >1000 mg/l, respectively.

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 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 200 mg/l was prepared by dissolving test chemical in reconstituted water. Thus, limit test was performed using a nominal test conc. of 200 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 >200 mg/l (with percent immobility as 4.0 at 100 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.

Another short term toxicity to aquatic invertebrate study was conducted for 48 hrs for assessing the effect of test chemical. The study was performed under static conditions using Daphnia spp. (Water flea) as a test organism.On the basis of the toxic effect of the chemical on mobility of the test organism Daphnia magna,the 48 hr median lethal concentration (LC50) value was determined to be >1000 mg/l.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/LC50 value can be expected to be in the range 200 to >1000 mg/l, respectively.

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/LC50 value can be expected to be in the range 200 to >1000 mg/l, respectively.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

200 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 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 200 mg/l was prepared by dissolving test chemical in reconstituted water. Thus, limit test was performed using a nominal test conc. of 200 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 simultaneously 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 >200 mg/l (with percent immobility as 4.0 at 100 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.

 

Another short term toxicity to aquatic invertebrate study was conducted for 48 hrs for assessing the effect of test chemical. The study was performed under static conditions using Daphnia spp. (Water flea) as a test organism. On the basis of the toxic effect of the chemical on mobility of the test organism Daphnia magna, the 48 hr median lethal concentration (LC50) value was determined to be >1000 mg/l. 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/LC50 value can be expected to be in the range 200 to >1000 mg/l, respectively.