2-[3-(1,3-dihydro-1,3,3-trimethyl-2H-indol-2-ylidene)prop-1-enyl]-1,3,3-trimethyl-3H-indolium chloride

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:

of read aross 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

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 study of aquatic invertebrate for the test chemical :
1.This study was designed to assess the toxic effects of the test compound on the test Daphnids.
2.To evalaute the toxicity on test material on aquatic invertebrate Daphnia magna

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (IUPAC name): 1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride
- Common name: Basic Red 12
- Molecular formula: C25H29N2.Cl
- Molecular weight: 392.971 g/mol
- Smiles notation: c12c(C(C)(C)C(\N1C)=C/C=C/C=1C(c3ccccc3[N+]1C)(C)C)cccc2.[ClH-]
- InChl: 1S/C25H29N2.ClH/c1-24(2)18-12-7-9-14-20(18)26(5)22(24)16-11-17-23-25(3,4)19-13-8-10-15-21(19)27(23)6;/h7-17H,1-6H3;1H/q+1;/p-1
- Substance type: Organic

Analytical monitoring:

not specified

Details on sampling:

1.PREPARATION AND APPLICATION OF TEST SOLUTION
The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 10 mg of the test substance in 100 ml of ADaM’s media. Achieving test concentrations of 100 mg/L, respectively.

2.The stock solution (100 g/L) was prepared by dissolving white powder in acetone. The test solutions of required concentrationas were prepared by mixing the stock solution of the test sample with reconstituted test water. The test solution were kept 10 min in ultrasonic bath.

Vehicle:

not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

1 . TEST ORGANISM
- Common name: Water flea
- Strain/clone: No data
- Source: Own breeding of Daphnia magna
- Age: No data
- Feeding during test : No feeding after hatching

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

QUARANTINE (wild caught)
- Duration: No data
- Health/mortality: No data

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

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

Limit test:

yes

Total exposure duration:

48 h

Hardness:

1) 145.5 mg of CaCO3

Test temperature:

1) and 2) 20 °C ± 2 °C

pH:

1) 7.1
2)Sample at concentration 100 mg/l: pH= 7.8 (changed to 7.7 during test)
Control 1: pH= 7.6 (did not change during test)
Control 1 + aceotone: pH=8 (changed to 8.1 during test)
Control 2: pH=7.8 (changed to 7.7 during test)
Control 2 + aceotone: pH=7.7 (did not changed during test)

Nominal and measured concentrations:

1) 100 mg/L
2)6, 10, 17, 29, 50 and 100 mg/L

Details on test conditions:

1 . TEST SYSTEM
- Test vessel: The test vessels used were all glass beaker having the water capacity of 25ml.
- Material, size, headspace, fill volume: 20 ml
- Aeration: No aeration during experiment
- No. of organisms per vessel: 10 Daphnids
- No. of vessels per concentration (replicates): No data
- No. of vessels per control (replicates): No data
- No. of vessels per vehicle control (replicates): No data
- Biomass loading rate: No data

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: No data
- Total organic carbon: No data
- Particulate matter: No data
- Metals: No data
- Pesticides: No data
- Chlorine: No data
- Alkalinity: No data
- Ca/mg ratio: No data
- Conductivity: No data
- Salinity: No data
- Culture medium different from test medium: No data
- Intervals of water quality measurement: No data

OTHER TEST CONDITIONS
- Adjustment of pH: No data
- Photoperiod: 16 hours light and 8 hours dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Determination or calculation of the median effective concen trations (EC 50) and observations ((immobility, pH, Temperature, dissolved oxygen content) ) of the symptoms were done after 24 and 48 hours.

VALIDITY OF THE TEST:
1. In the control, including the control containing the solubilising agent, not more that 10 percent of the daphnids should have been immobiliz ed.
2. The dissolved oxygen concentration at the end of the test should be 3 mg/l in control and test vessels.

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

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr2O7) for Read across 2

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: Read across 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

29.8 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Read across 2

Details on results:

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

Validity criteria fulfilled:

not specified

Conclusions:

The test chemical 1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride is likely to be toxic to Daphnia magna in the dose range of 29.8 to 100 mg/l

Executive summary:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the short term toxicity of aquatic invertebrate of the test chemical 1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride (6320 -14 -5).The studies are as mentioned below:

1.Daphnia sp., Acute Immobilization Test according to OECD Guideline 202 was conducted for test material. The nominal concent ration selected for the experiment was 100 mg/L and test Daphnids were exposed to this concentration for 48hours. The Effective concentrations EC50 was found to be 100 mg/L. Also the substance was found to be not rapidly degradable.Thus according to CLP Criteria for aquatic classification of the substance, it is concluded that test material exhibits toxicity to aquatic invertebrate (Daphnia magna) in the chronic 3 classification category.

EC50 (48 hours) Experimental = 100 mg/L

2.Determination of the inhibition of the mobility of daphnids was carried out with the substance,according to OECD Guideline 202.

The test substance was tested at the concentrations 0, 6, 10, 17, 29, 50 and 100 mg/L. Effects on immobilisation were observed for 48 hours. The median effective concentration (EC50) for the test substance,1-(3-chloropropyl)-1,3-dihydro-2H-benzimidazol-2-one, in Daphnia magna was determined to be 29.8 mg/L for immobilisation effects.

This value indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified as Aquatic Chronic category 3 as per the CLP criteria.

Thus, based on the above summarised studies, 1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride and it’s functionally similar read across substance, it can be concluded that effect concetration value is in the range of 29 to 100 mg/

L. Thus, comparing this value with the criteria of CLP regulation,

1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride can be classified

for short term toxicity for aquatic invertebrate .Hence,based on the data available for functionally similar read across, test chemical1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride can be classified as Aquatic Chronic category 3 as per the CLP criteria.

.

Description of key information

Short-term toxicity to aquatic invertebrates:

Data available for the closely related read across chemicals has been reviewed to determine the short term toxicity of aquatic invertebrate of the test chemical 1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride (6320 -14 -5).The studies are as mentioned below:

1.Daphnia sp., Acute Immobilization Test according to OECD Guideline 202 was conducted for test material. The nominal concent ration selected for the experiment was 100 mg/L and test Daphnids were exposed to this concentration for 48hours. The Effective concentrations EC50 was found to be 100 mg/L. Also the substance was found to be not rapidly degradable.Thus according to CLP Criteria for aquatic classification of the substance, it is concluded that test material exhibits toxicity to aquatic invertebrate (Daphnia magna) in the chronic 3 classification category.

EC50 (48 hours) Experimental = 100 mg/L

2.Determination of the inhibition of the mobility of daphnids was carried out with the substance,according to OECD Guideline 202.

The test substance was tested at the concentrations 0, 6, 10, 17, 29, 50 and 100 mg/L. Effects on immobilisation were observed for 48 hours. The median effective concentration (EC50) for the test substance,1-(3-chloropropyl)-1,3-dihydro-2H-benzimidazol-2-one, in Daphnia magna was determined to be 29.8 mg/L for immobilisation effects.

This value indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified as Aquatic Chronic category 3 as per the CLP criteria.

Thus, based on the above summarised studies, 1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride and it’s functionally similar read across substance, it can be concluded that effect concetration value is in the range of 29 to 100 mg/

L. Thus, comparing this value with the criteria of CLP regulation,

1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride can be classified for short term toxicity for aquatic invertebrate .Hence,based on the data available for closely related read across, test chemical1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride can be classified as Aquatic Chronic category 3 as per the CLP 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 invertebrates:

Data available for the closely related read across chemicals has been reviewed to determine the short term toxicity of aquatic invertebrate of the test chemical 1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride (6320 -14 -5).The studies are as mentioned below:

1.Daphnia sp., Acute Immobilization Test according to OECD Guideline 202 was conducted for test material. The nominal concent ration selected for the experiment was 100 mg/L and test Daphnids were exposed to this concentration for 48hours. The Effective concentrations EC50 was found to be 100 mg/L. Also the substance was found to be not rapidly degradable.Thus according to CLP Criteria for aquatic classification of the substance, it is concluded that test material exhibits toxicity to aquatic invertebrate (Daphnia magna) in the chronic 3 classification category.

EC50 (48 hours) Experimental = 100 mg/L

2.Determination of the inhibition of the mobility of daphnids was carried out with the substance,according to OECD Guideline 202.

The test substance was tested at the concentrations 0, 6, 10, 17, 29, 50 and 100 mg/L. Effects on immobilisation were observed for 48 hours. The median effective concentration (EC50) for the test substance,1-(3-chloropropyl)-1,3-dihydro-2H-benzimidazol-2-one, in Daphnia magna was determined to be 29.8 mg/L for immobilisation effects.

This value indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified as Aquatic Chronic category 3 as per the CLP criteria.

Thus, based on the above summarised studies, 1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride and it’s functionally similar read across substance, it can be concluded that effect concetration value is in the range of 29 to 100 mg/

L. Thus, comparing this value with the criteria of CLP regulation,

1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride can be classified

for short term toxicity for aquatic invertebrate .Hence,based on the data available for closely related read across, test chemical1,3,3-trimethyl-2-[(1E)-3-[(2E)-1,3,3-trimethyl-2,3-dihydro-1H-indol-2-ylidene]prop-1-en-1-yl]-3H-indol-1-ium chloride can be classified as Aquatic Chronic category 3 as per the CLP criteria.

.