[4-[[4-anilino-1-naphthyl][4-(dimethylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium acetate

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:

guideline study

Justification for type of information:

Data is from UERL lab.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Principles of method if other than guideline:

This study was designed to assess the toxic effects of the test compound on the ZEBRA FISH(Danio rerio) .Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes each. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. The experimental phase of the main study started at 27-02-207 and lasted for a period of 96 hrs.

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material: N-(4-{[4-(dimethylamino)phenyl][4-(methylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium acetate- Molecular formula: C24 H28 N3.C2 H3 O2- Molecular weight: 417.5499 g/mol- Smiles notation: CC(=O)[O-].CNc1ccc(cc1)C(=C2C=CC(=[N+](C)C)C=C2)c3ccc(cc3)N(C)C- InChl : 1S/C24H27N3.C2H4O2/c1-25-21-12-6-18(7-13-21)24(19-8-14-22(15-9-19)26(2)3)20-10-16-23(17-11-20)27(4)5;1-2(3)4/h6-17H,1-5H3;1H3,(H,3,4)- Substance type: Organic- Physical state: liquid - Batch no: KCP/FS//1701/17- Date of manufacturing : 01-04-2017- Storage Conditions: Ambient Temp (23 to 27 °C) √ (Store in a cool place. Keep containers tightly closed in a dry and well ventilated place)

Analytical monitoring:

not specified

Vehicle:

no

Details on test solutions:

The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 100 mg of the test substance in 100 ml of potable water (passed through reverse osmosis system) with 1 hr.continuous stirring for achieving test concentrations of 1g/L. Hence, concentrations are 0.3125mg/L,0.625mg/L,1.25mg/L,2.5mg/L,5mg/L ,respectively.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM- Common name:ZEBRA FISH- Source:UNIQUE AQUARIUM, NAGPUR- Length at study initiation (length definition, mean, range and SD):average 1.94 cm- Weight at study initiation (mean and range, SD):0.1494 g (average)ACCLIMATION- Acclimation period:7 days- Type and amount of food during acclimation:Standard brand feed- Feeding frequency during acclimation:Daily once- Health during acclimation (any mortality observed):During last 48 hours = 0FEEDING DURING TEST- Food type:Standard brand feed- Frequency:Withdrawal of food 1 day before and during exposure

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

148.5 mg of CaCO3

Test temperature:

23 °C

pH:

7.12

Dissolved oxygen:

7.8 mg/l

Nominal and measured concentrations:

0.3125mg/L, 0.625mg/L,1.25mg/L, 2.5mg/L, 5mg/L

Details on test conditions:

TEST SYSTEM - Test vessel:The test vessels used were all plastic aquaria having the water capacity of 5liters. - Material, size, headspace, fill volume: 2 liters - No. of organisms per vessel:8 fishes OTHER TEST CONDITIONS - Photoperiod:16 hours light and 8 hours dark

Reference substance (positive control):

not specified

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.312 - < 0.625 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Toxic

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

0.312 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Toxic

Key result

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

0.625 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Toxic

Details on results:

No data available

Results with reference substance (positive control):

No data available

Reported statistics and error estimates:

No data available

Sublethal observations / clinical signs:

Mortality and symptoms:

Nominal Concentration (mg/l)	Number of fish		Mortality (cumulated) after
			3 h		6 h		24h		48h		72h		96h
			V1	V2	V1	V2	V1	V2	V1	V2	V1	V2	V1	V2
Control	4	4	-	-	-	-	-	-	-	-	-	-	-	-
0.3125	4	4	-	-	-	-	-	-	-	-	-	-	-	-
0.625	4	4	-	-	-	-	4	4	4	4	4	4	4	4
1.25	4	4	-	-	-	-	4	4	4	4	4	4	4	4
2.5	4	4	-	-	4	4	4	4	4	4	4	4	4	4
5	4	4	4	4	4	4	4	4	4	4	4	4	4	4

 

Visible Symptoms:

Nominal Concentration (mg/l)	Number of fish		symptom (cumulated) after
			3 h		6 h		24h		48h		72h		96h
			V1	V2	V1	V2	V1	V2	V1	V2	V1	V2	V1	V2
Control	4	4	NS4	NS4	NS4	NS4	NS4	NS4	NS4	NS4	NS4	NS4	NS4	NS4
0.3125	4	4	NS4	NS4	NS4	NS4	NS4	NS4	NS4	NS4	NS4	NS4	NS4	NS4
0.625	4	4	NS4	NS4	NS4	NS4	M4	M4	-	-	-	-	-	-
1.25	4	4	M4	M4	M4	M4	M4	M4	-	-	-	-	-	-
2.5	4	4	M4	M4	IS4	IS4	-	-	-	-	-	-	-	-
5	4	4	IS4	IS4	-	-	-	-	-	-	-	-	-	-

Explanation of symptoms:

A: apathy

N: Narcotic-like state

C: Swimming near water surface

M: Slow movements compared to control

NS: No abnormal symptoms

S: Surfacing behaviour

L: Loss of equilibrium

SM: Shimming movement

H1: Hyperactivity

H2: Hypo activity

B: Bending behavior

F: Fin rot

IS: Invert Swimming

SB: Swimming behaviour

RF: Respiratory Function

P: Pigmentation

Validity criteria fulfilled:

yes

Conclusions:

Based on mortality of fishes due to the exposure of chemical [4-[[4-anilino-1-naphthyl][4 -(dimethylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium acetate at nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for CAS No.83803-79-6 on Zebra fish(Danio rerio)was determine to be >0.3125 mg/L but <0.625 mg/L.

Executive summary:

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for (test item name) [4-[[4-anilino-1-naphthyl][4 -(dimethylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene] dimethylammonium acetate. The test solution was prepared by dissolving 100 mg of the test substance in 100 ml of potable water (passed through reverse osmosis system) with 1 hr.continuous stirring for achieving test concentrations of 1g/L. The nominal concentration selected for the experiment were 0.3125mg/L,0.625mg/L,1.25mg/L,2.5mg/L,5mg/Land test fish were exposed to these concentration for 96 hours. After 96hrs of exposure LC0, LC50 and LC100 was observed.

The lethal concentrations LC50 was determine to be >0.3125 mg/L but <0.625 mg/L

LC0 (96 hours) (highest loading at which no mortality was observed) = 0.3125 mg/L

LC50 (96 hours) Experimental = >0.3125 mg/L but <0.625 mg/L

LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 0.625 mg/L

Description of key information

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for (test item name) [4-[[4-anilino-1-naphthyl][4 -(dimethylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene] dimethylammonium acetate. The test solution was prepared by dissolving 100 mg of the test substance in 100 ml of potable water (passed through reverse osmosis system) with 1 hr.continuous stirring for achieving test concentrations of 1g/L. The nominal concentration selected for the experiment were 0.3125mg/L,0.625mg/L,1.25mg/L,2.5mg/L,5mg/Land test fish were exposed to these concentration for 96 hours. After 96hrs of exposure LC0, LC50 and LC100 was observed.

The lethal concentrations LC50 was determine to be >0.3125 mg/L but <0.625 mg/L

LC0 (96 hours) (highest loading at which no mortality was observed) = 0.3125 mg/L

LC50 (96 hours) Experimental = >0.3125 mg/L but <0.625 mg/L

LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 0.625 mg/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.312 mg/L

Additional information

Based on the various predicted data for the target chemical study have been reviewed to determine the toxic nature of target chemical N-(4-{[4-(dimethylamino)phenyl][4-(methylamino)phenyl]methylene} cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium acetate (83803 -79 -6). The studies are as mentioned below:

 

In the first experimental key study for the target chemical (83803-79-6) from UERL lab, 2016, toxicity was measured on the fish. Fish Acute Toxicity test according to OECD Guideline 203 was conducted for (test item name) [4-[[4-anilino-1-naphthyl][4 -(dimethylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene] dimethylammonium acetate. The test solution was prepared by dissolving 100 mg of the test substance in 100 ml of potable water (passed through reverse osmosis system) with 1 hr.continuous stirring for achieving test concentrations of 1g/L. The nominal concentration selected for the experiment were 0.3125mg/L,0.625mg/L,1.25mg/L,2.5mg/L,5mg/Land test fish were exposed to these concentration for 96 hours. After 96hrs of exposure LC0, LC50 and LC100 was observed.

The lethal concentrations LC50 was determine to be >0.3125 mg/L but <0.625 mg/L

LC0 (96 hours) (highest loading at which no mortality was observed) = 0.3125 mg/L

LC50 (96 hours) Experimental = >0.3125 mg/L but <0.625 mg/L

LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 0.625 mg/L

 

 

Similarly in the second supporting study for the target chemical (83803-79-6) from Department of Toxicology BASF SE, 1989. Study was conducted to determine the toxic nature of chemical N-(4-{[4-(dimethylamino)phenyl][4-(methylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium acetate on the growth of fish Leuciscus idus. Test was performed according to the OECD guideline. The doses were chosen based on two range finding studies between 0.3 and 1 mg/l) the concentrations, spaced by a factor of about 2.2. Control, 0.01, 0,022, 0.046, 0.1, 0.215, 0464, 1.0, 2.15 mg/L concentration was taken in the study. In the form of an aqueous solution (0.1 g/100 ml) the product was added to the test water, subsequently the fish were placed into the aquaria. Leuciscus idus (golden variety) used as a test organism collected from Fischzucht Paul Eggers, 2354 Hohenwestedt, Germany. Fish does not feed during test. Open glass aquaria having dimeter 30 x 22 x 24 cm, was used having the capacity of 10 liter. 2.8 g fish /L was added. 10 organisms per vessel was used. Chloracetamid added as a reference substance. After the exposure of 96hrs LC50 observed on the basis of Apathy and narcotic-like state was determine at test concentration 2.15 mg/L after 1 hour. In reference substance Chloracetamid the LC50 was 32 mg/L at 48 hr. Based on the mortality LC50 was 0.158 mg/l and the LC100 was 0.0464 mg/l.

Based on the LC50, chemical was consider as toxic and can be consider to be classified as aquatic acute 1 category as per the CLP classification criteria.