2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3-trimethyl-3H-indolium chloride

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from peer reviewed journal

Justification for type of information:

Data is from peer reviewed journal

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

Short term toxicity test to rainbow trout, bluegill and sea lamprey was conducted for 24 hrs.

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (IUPAC name): 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride
- Common name: Astrazonrot 6B
- Molecular formula: C24H30ClN2Cl
- Molecular weight: 417.421 g/mol
- Smiles notation: C1(=[N+](c2ccccc2C1(C)C)C)\C=C/c1c(cc(N(CCCl)CC)cc1)C.[ClH-]
- InChl: 1S/C24H30ClN2.ClH/c1-6-27(16-15-25)20-13-11-19(18(2)17-20)12-14-23-24(3,4)21-9-7-8-10-22(21)26(23)5;/h7-14,17H,6,15-16H2,1-5H3;1H/q+1;/p-1
- Substance type: Organic
- Physical state: Solid

Analytical monitoring:

not specified

Details on sampling:

- Concentrations: 5 mg/l (equal to 5 ppm)

- Sampling method: Chemical was weighed in calibrated weighing bottles to the nearest milligram on a Volland Speedigram balance. Five cubic centimeters of the indicated solvent was added to the test chemical. Each concentrated mixture was next added to a predetermined volume of water (as required by actual weight of test chemical and desired conc.) and agitated with a Power-Stir to produce a more dilute solution.

Emulsions or suspensions of insoluble compounds were made with the aid of a Waring blender.

These prepared solutions, emulsions or suspensions were added to the test containers in which the experimental animals had already been placed. The resultant volume in each test jar varied from 5800 to 6200 cc.

Vehicle:

yes

Remarks:

water, acetone or ethyl alcohol (absolute) was used as a vehicle.

Test organisms (species):

other: Test organism used for the study were Salmon gairdnerii, Lepomis macrochius and Petromyzon marinus

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout, Bluegill and Sea Lamprey

- Source:
Sea lamprey (Petromyzon marinus )- Ocqueoc River, Presque Isle County, Michigan.

Rainbow trout (Salmo gairdnerii) and Bluegill (Lepomis macrochius)- Test fishes were obtained from the stocks of local state and Federal fish hatcheries.

- Length at study initiation (length definition, mean, range and SD):
Sea lamprey (Petromyzon marinus )- 3 to 5 inches in length.

Rainbow trout (Salmo gairdnerii) and Bluegill (Lepomis macrochius)- fingerling size, 4 inches or slightly less in length.

- Method of breeding: Larval lampreys were collected by means of an electric shocker in the Ocqueoc River, Presque Isle County, Michigan, and were held in running water in aquaria and small “races” under conditions which simulated their natural stream habitat.

Test fishes (Rainbow trout and Bluegill) were obtained from the stocks of local state and Federal fish hatcheries and were held in large raceways. These specimens were maintained in the best possible physical condition until used in the laboratory.

Test type:

static

Water media type:

freshwater

Total exposure duration:

24 h

Test temperature:

12.77 ᵒC (55ᵒF)

pH:

7.5 to 8.2 pH

Nominal and measured concentrations:

Nominal concentrations

Details on test conditions:

TEST SYSTEM
- Test vessel: glass battery jar containing 5 liters of water.
- Material, size, headspace, fill volume: 10-liter
- Aeration: the jars were provided with aeration through standard stone air-breakers.
- No. of organisms per vessel: six organisms
- No. of vessels per concentration (replicates): 12 jars were included in each trough
- No. of vessels per control (replicates): 1 control jar was included in each trough

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Water used in all tests was drawn from a supply pumped directly from Hammond Bay of Lake, Huron.

The suction line intake of this pumping system was located 250 feet offshore at 3 depth of about 9 feet. Water from this source was consistently clear and of a relatively unIform quality.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Observations of each test were measured approx. six times, at various intervals, during the 24 hr test period. At each observation, the condition of every test specimen was determined and recorded. Chronological histories were thus obtained of any symptoms of illness and the occurrence of death.

TEST CONCENTRATIONS
- Test concentrations: 5 mg/l (equal to 5 ppm)

Reference substance (positive control):

not specified

Key result

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

not specified

Validity criteria fulfilled:

not specified

Conclusions:

No effects were observed when the test organisms sea lamprey, bluegill and rainbow trout were exposed to the test chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride for 24 hrs. Thus, the EC0 value was determined to be 5 mg/l.

Executive summary:

Short term toxicity test to rainbow trout, bluegill and sea lamprey was carried out for 24 hrs. Test organisms Sea lamprey (Petromyzon marinus), Rainbow trout (Salmo gairdnerii) and Bluegill (Lepomis macrochius) were used for the study. The study was performed under static conditions for 24 hrs. The test chemical2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride (CAS no. 6441-82-3) conc. used for the study was 5 mg/l. Larval lampreys were collected by means of an electric shocker in the Ocqueoc River, Presque Isle County, Michigan, and were held in running water in aquaria and small “races” under conditions which simulated their natural stream habitat.Test fishes (Rainbow trout and Bluegill) were obtained fromthe stocks of local state and Federal fish hatcheriesand were held in large raceways. These specimens were maintained in the best possible physical condition until used in the laboratory.

The aggregate test animals available, usually six in number, were placed together in a 10-literglass battery jar containing 5 liters of water. The jars were provided with aeration through standard stone air-breakersand were maintained at a constant temperature by immersion in specially constructed constant temperature troughs. Water temperature was maintained within the limits of ± 1.0 °F.Twelve of these test jars (each containing a substance being assayed) were included with one control jar in each trough. Fish and larvae in the control jar were exposed only to the water and physical conditions of the typical test container.Test condition contains dissolved oxygen from 8.6 to 13.7 ppm and free CO₂from 5.0 to 9.0 ppm. Observations of each test were measured approx. six times, at various intervals, during the 24 hr test period. At each observation, the condition of every test specimen was determined and recorded. Chronological histories were thus obtained of any symptoms of illness and the occurrence of death.No effects were observed when the test organisms sea lamprey, bluegill and rainbow trout were exposed to the test chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride for 24 hrs. Thus, the EC0 value was determined to be 5 mg/l.

Description of key information

Short term toxicity test to rainbow trout, bluegill and sea lamprey was carried out for 24 hrs (Vernon C. Applegate et. al; 1957). Test organisms Sea lamprey (Petromyzon marinus), Rainbow trout (Salmo gairdnerii) and Bluegill (Lepomis macrochius) were used for the study. The study was performed under static conditions for 24 hrs. The test chemical2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride (CAS no. 6441-82-3) conc. used for the study was 5 mg/l. Larval lampreys were collected by means of an electric shocker in the Ocqueoc River, Presque Isle County, Michigan, and were held in running water in aquaria and small “races” under conditions which simulated their natural stream habitat.Test fishes (Rainbow trout and Bluegill) were obtained fromthe stocks of local state and Federal fish hatcheriesand were held in large raceways. These specimens were maintained in the best possible physical condition until used in the laboratory.

The aggregate test animals available, usually six in number, were placed together in a 10-literglass battery jar containing 5 liters of water. The jars were provided with aeration through standard stone air-breakersand were maintained at a constant temperature by immersion in specially constructed constant temperature troughs. Water temperature was maintained within the limits of ± 1.0 °F.Twelve of these test jars (each containing a substance being assayed) were included with one control jar in each trough. Fish and larvae in the control jar were exposed only to the water and physical conditions of the typical test container.Test condition contains dissolved oxygen from 8.6 to 13.7 ppm and free CO₂from 5.0 to 9.0 ppm. Observations of each test were measured approx. six times, at various intervals, during the 24 hr test period. At each observation, the condition of every test specimen was determined and recorded. Chronological histories were thus obtained of any symptoms of illness and the occurrence of death.No effects were observed when the test organisms sea lamprey, bluegill and rainbow trout were exposed to the test chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride for 24 hrs. Thus, the EC0 value was determined to be 5 mg/l.

Additional information

Experimental study for the target chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride (CAS No. 6441-82-3) and various supporting weight of evidence studies for its structurally similar read across substanceswere reviewed to summarize the following information:

 

In an experiment key study from peer reviewed journal (Vernon C. Applegate et. al; 1957), Short term toxicity test to rainbow trout, bluegill and sea lamprey was carried out for 24 hrs. Test organisms Sea lamprey (Petromyzon marinus), Rainbow trout (Salmo gairdnerii) and Bluegill (Lepomis macrochius) were used for the study. The study was performed under static conditions for 24 hrs. The test chemical2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride (CAS no. 6441-82-3) conc. used for the study was 5 mg/l. Larval lampreys were collected by means of an electric shocker in the Ocqueoc River, Presque Isle County, Michigan, and were held in running water in aquaria and small “races” under conditions which simulated their natural stream habitat.Test fishes (Rainbow trout and Bluegill) were obtained fromthe stocks of local state and Federal fish hatcheriesand were held in large raceways. These specimens were maintained in the best possible physical condition until used in the laboratory.

The aggregate test animals available, usually six in number, were placed together in a 10-literglass battery jar containing 5 liters of water. The jars were provided with aeration through standard stone air-breakersand were maintained at a constant temperature by immersion in specially constructed constant temperature troughs. Water temperature was maintained within the limits of ± 1.0 °F.Twelve of these test jars (each containing a substance being assayed) were included with one control jar in each trough. Fish and larvae in the control jar were exposed only to the water and physical conditions of the typical test container.Test condition contains dissolved oxygen from 8.6 to 13.7 ppm and free CO2from 5.0 to 9.0 ppm. Observations of each test were measured approx. six times, at various intervals, during the 24 hr test period. At each observation, the condition of every test specimen was determined and recorded. Chronological histories were thus obtained of any symptoms of illness and the occurrence of death.No effects were observed when the test organisms sea lamprey, bluegill and rainbow trout were exposed to the test chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride for 24 hrs. Thus, the EC0 value was determined to be 5 mg/l.

 

In a supportingstudy from authoritative database (ECOTOX database, 2018) of the read across chemical 2-(2-{4-[(2-chloroethyl)(methyl)amino]phenyl}vinyl)-1,3,3-trimethyl-3H-indolium chloride (CAS no. 3648-36-0),short term toxicity to Leuciscus idus (Ide, Silver or Golden Orfe) study was carried out for 48 hrs. The study was based on the effects of the read across compound 2-(2-{4-[(2-chloroethyl)(methyl)amino]phenyl}vinyl)-1,3,3-trimethyl-3H-indolium chloride (CAS no. 3648-36-0) on Leuciscus idus in a static freshwater system at a temperature of 10 to 13°C and pH 7.2 to 7.4, respectively. Hardness of water is 120 to 140 mg/l CaCO3 and dissolved oxygen is ˃4 mg/l, respectively. Leuciscus idus (Ide, Silver or Golden Orfe) was used as a test organism. Based on effect on mortality of the test organism Leuciscus idus, the 48 hr LC50 value was determined to be 40 mg/l, respectively. Thus, based on the LC50 value, it can be concluded that the substance 2-(2-{4-[(2-chloroethyl)(methyl)amino]phenyl}vinyl)-1,3,3-trimethyl-3H-indolium chloride can be considered as toxic to aquatic organisms and thus can be considered to be classified in aquatic chronic category 3 as per the CLP classification criteria.

 

Another short term toxicity to Leuciscus idus (Ide, Silver or Golden Orfe) study was carried out for 48 hrs (ECOTOX database, 2018). The study was based on the effects of the read across compound 2-(2-{4-[(2-chloroethyl)(methyl)amino]phenyl}vinyl)-1,3,3-trimethyl-3H-indolium chloride (CAS no. 3648-36-0) on Leuciscus idus in a static freshwater system at a temperature of 13°C and pH 7.0 to 8.0, respectively. Hardness of water is 270 mg/l CaCO3 and dissolved oxygen is ˃5 mg/l, respectively. Leuciscus idus (Ide, Silver or Golden Orfe) was used as a test organism. Based on effect on mortality of the test organism Leuciscus idus, the 48 hr LC50 value was determined to be in the range 38 to 75 mg/l, respectively.Thus, based on the LC50 value, it can be concluded that the substance 2-(2-{4-[(2-chloroethyl)(methyl)amino]phenyl}vinyl)-1,3,3-trimethyl-3H-indolium chloride can be considered as toxic to aquatic organisms and thus can be considered to be classified in aquatic chronic category 3 as per the CLP classification criteria.

 

For the same read across chemical compound 2-(2-{4-[(2-chloroethyl)(methyl)amino]phenyl}vinyl)-1,3,3-trimethyl-3H-indolium chloride (CAS no. 3648-36-0), short term toxicity to Oncorhynchus mykiss (Rainbow Trout) study was carried out for 48 hrs (ECOTOX database, 2018). The study was based on the effects of the read across compound 2-(2-{4-[(2-chloroethyl)(methyl)amino]phenyl}vinyl)-1,3,3-trimethyl-3H-indolium chloride on Oncorhynchus mykiss in a static freshwater system at a temperature of 13°C and pH 7.0 to 7.4, respectively. Hardness of water is 180 mg/l CaCO3 and dissolved oxygen is ˃7 mg/l, respectively. Oncorhynchus mykiss (Rainbow Trout) was used as a test organism. Based on effect on mortality of the test organism Oncorhynchus mykiss, the 48 hr LC50 value was determined to be 80 mg/l, respectively. Thus, based on the LC50 value, it can be concluded that the substance 2-(2-{4-[(2-chloroethyl)(methyl)amino]phenyl}vinyl)-1,3,3-trimethyl-3H-indolium chloride can be considered as toxic to aquatic organisms and thus can be considered to be classified in aquatic chronic category 3 as per the CLP classification criteria.

 

Thus, based on the overall reported results for target chemical 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride (from peer reviewed journal) and for its read across substance (from authoritative database ECOTOX),it can be concluded that the EC0 value of the test substance 2-[2-[4-[(2-chloroethyl)ethylamino]-o-tolyl]vinyl]-1,3,3- trimethyl-3H-indolium chloride was determined to be 5 mg/l.