[4-[[4-(dimethylamino)phenyl][4-(methylamino)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

Study period:

25-04-2017 to 29-04-2017

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 (as cited in study report): N-(4-{[4-(dimethylamino)phenyl][4-(methylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium acetate
- Molecular formula: C24H28N3C2H3O2
- 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: Greenish lustrous Liquid
- Batch No: KCP/FS//170/17
- Stability: Stable in recommended storage conditions
- Solubility in Water: 102950 mg/L

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 1 g/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 = 0

FEEDING 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 and 100 mg/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)

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
5	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

IS4

IS4

-

-

-

-

-

-

1.25

4

4

NS4

NS4

NS4

NS4

IS4

IS4

-

-

-

-

-

-

2.5

4

4

NS4

NS4

NS4

NS4

IS4

IS4

-

-

-

-

-

-

5

4

4

NS4

NS4

NS4

NS4

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 nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for CAS No.84434-47-9 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 -(dimethylamino)phenyl][4-(methylamino)phenyl]methylene] cyclohexa-2,5-dien- 1-ylidene]dimethylammonium acetate 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.

The lethal concentrations LC50 was found 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 -(dimethylamino)phenyl][4-(methylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene] dimethylammonium acetate 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.

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

Three studies including predicted data from experimental data and from authorative database and also from validated tools for short term fish toxicity endpoint of target chemical N-(4-{[4 -(dimethylamino)phenyl][4-(methylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium acetate (Cas no. 84434-47-9) with relevant read across chemical which is structurally similar to target were summarized as follows:

First key experimental study suggest Fish Acute Toxicity test according to OECD Guideline 203 was conducted for (test item name) [4-[[4 -(dimethylamino)phenyl][4-(methylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium acetate 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. The lethal concentrations LC50 was found 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 and LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 0.625 mg/L 

Similarly above experimental result supported by prediction using the aquatic toxicity module i.e ACD/LabsI-Lab 2.0; Algorithm Version: v5.0.0.184, the lethal concentration (LC50) of the test substance N-(4-{[4-(dimethylamino)phenyl][4-(methylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium acetate was predicted to be 0.12 mg/l on fish species Fathead minnow (Pimephales promelas).

The Reliability Index (RI) of the prediction was reported to be 0.6(As per model substance have moderate and high reliability if RI > 0.5). Thus reported RII value indicate it has moderate reliability. Based on the LC50 value, it can be concluded that the substance N-(4 -{[4 -(dimethylamino)phenyl](phenyl)methylene}cyclohexa-2,5 -dien-1 -ylidene)-N-methylmethanaminium chloride can be considered as toxic to aquatic organisms and thus classified in aquatic category 1 as per the CLP criteria.

 

In the third supportingstudy experiment data of read across chemical N-(4-{[4-(dimethylamino)phenyl](phenyl)methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride (Cas no. 569 -64 -2) wherein acute fish toxicity study was carried out to determine the lethal concentration (LC50) of test substance on fish for exposure period of 96 hrs. Test organism used was Morone saxatilis (Striped Bass) The Lethal concentration to 50% of test organisms (LC50) on the basis of mortality effect was observed to be 0.138 mg/l for exposure period of 96 hrs. (Ecotox database by US EPA)

 

Thus available studies for short term fish toxicity give common conclusion as test chemical N-(4-{[4-(dimethylamino)phenyl][4-(methylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)- N-methylmethanaminium acetate (Cas no. 84434-47-9) was toxic to fish and thus classified in aquatic category 1 as per the CLP criteria.