Reaction mass of 4-(2,6,6-trimethylcyclohex-1-ene-1-yl)-but-3-ene-2-one and 4-(2,6,6-trimethylcyclohex-2-ene-1-yl)-but-3-ene-2-one

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:

Experimental test result performed according to the guideline.

Qualifier:

according to

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Principles of method if other than guideline:

To study the effects of test chemical on fish, test was carried out according to the OECD Guideline 203 (Fish, Acute Toxicity Test) .

GLP compliance:

no

Analytical monitoring:

no

Details on sampling:

The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 50 mg & 75 mg of the test substance in 10 liters deionized water with continuous stirring for achieving test concentrations of 5 mg/L and 7.5 mg/L, respectively.

Vehicle:

no

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Test species/strain : Zebra Fish (Danio rerio)
Animal breeder/supplier : Ashish Fish Center, Dharampeth, Nagpur, MS (India)
Age of the animals : 3-4 months
Weight/length and physical condition at the initiation of the study:
Average weight: 0.116 g (approx.)
Average length: 2.3 cm
Physical conditions: No abnormalities
Housing : The fishes were kept in a static tank in tap water, under natural conditions for 15 days along with proper feed and aeration.
Aeration : Aeration in test vessels was provided 1 day before the start of experiment
Diet : Standard brand feed, once daily
Duration of housing : 15 days
Photoperiod : 12 hours light and 12 hours darkness
Oxygen content : 7.4

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

29°C

pH:

between 6-7

Dissolved oxygen:

7.4

Nominal and measured concentrations:

5 mg/L & 7.5 mg/L

Details on test conditions:

The test vessels used were all glass bowl aquaria of 12” each

Test concentrations:
Test concentrations selected for the experiment are based upon the available data and hence the range finding study was planned using two test substance concentrations i.e., 5 mg/L and 7.5 mg/L.

Range finding test:
Range finding test was conducted with the two test concentrations, viz., 5 mg/L and 7.5 mg/L. But at 7.5 mg/L one mortality was observed within 24 hrs. Therefore, the experiment was continued for both the concentration, i.e., 5 mg/L and 7.5 mg/L in which no mortality was found. Observations (mortality, visible symptoms, pH, Temperature, dissolved oxygen content) were recorded after 24 hours, 48 hours, 72 hours and 96 hours of the start of the experiment.

Reference substance (positive control):

not specified

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

7.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: Other details not known

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 7.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: Other details not known

Mortality and symptoms:

Nominal conc. (mg/l)	Number of fish	Mortality (cumulated) after
		24h	48h	72h	96h
0 (control)	8	0	0	0	0
5	8	0	0	0	0
7.5	8	1	0	0	0

 

Nominal conc. (mg/l)	Number of fish	Symptoms after
		24h	48h	72h	96h
0 (control)	8	NS8	NS8	NS8	NS8
5	8	NS8	NS8	NS8	NS8
7.5	8	NS7	NS7	NS7	NS7

 

Explanation of symptoms:

A: apathy

N: narcotic-like state

C: swimming near water surface

D: swimming at the bottom of bowl

M: Slow movements compared to control

NS: No abnormal symptoms

Number behind symbol for symptom= number of affected fish

pH and oxygen content:

Nominal conc. (mg/l)	pH readings after
	24h	48h	72h	96h
0 (control)	7.2	6.5	6.2	6.3
5	6.5	6.3	5.9	5.7
7.5	6.1	5.9	5.7	5.4

Nominal conc. (mg/l)	Oxygen content (mg/l) after
	24h	48h	72h	96h
0 (control)	7.6	6.7	6.5	6.1
5	6.5	6.3	6.2	6.0
7.5	6.7	6.3	6.3	6.1

 

Test temperature:

Nominal conc. (mg/l)	Temperature (°C) after
	24h	48h	72h	96h
0 (control)	29	30	31	31
5	29	30	31	31
7.5	29	30	31	31

Validity criteria fulfilled:

yes

Conclusions:

The 96 hrs Lethal concentration (LC50) value of test chemical on Danio rerio fish was observed at dose concentration of > 7.5 mg/L. As no effect were observed at 7.5 mg/l nominal concentration.

Executive summary:

A 96 h study was conducted for test chemical on Danio rerio, at the test concentrations of 5mg/L, & 7.5 mg/L. In5mg/L, & 7.5 mg/L, the fishes were found to be freely swimming in the bowl aquaria. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 50 mg & 75 mg of the test substance in 10 liters deionized water with continuous stirring for achieving test concentrations of 5 mg/L and 7.5 mg/L, respectively. No mortalities were found in the control aquaria. Hence the LC50 value was considered to be > 7.5 mg/L and the LC0 value was determine to be 7.5 mg/l. Thus considering CLP Criteria for aquatic classification of the substance , it is concluded that test chemical exhibits short term toxicity to fish (Danio rerio) and classified in the chronic 2 classification category. But as the chemical was readily biodegradable in water, thus on that basis chemical consider to be nontoxic and not classified as per the CLP classification criteria.

Description of key information

A 96 h study was conducted for test chemical on Danio rerio, at the test concentrations of 5mg/L, & 7.5 mg/L. In5mg/L, & 7.5 mg/L, the fishes were found to be freely swimming in the bowl aquaria. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 50 mg & 75 mg of the test substance in 10 liters deionized water with continuous stirring for achieving test concentrations of 5 mg/L and 7.5 mg/L, respectively. No mortalities were found in the control aquaria. Hence the LC50 value was considered to be > 7.5 mg/L and the LC0 value was determine to be 7.5 mg/l. Thus considering CLP Criteria for aquatic classification of the substance , it is concluded that test chemical exhibits short term toxicity to fish (Danio rerio) and classified in the chronic 2 classification category.

Key value for chemical safety assessment

LC50 for freshwater fish:

7.5 mg/L

Additional information

Based on the various experimental data for the test chemical study have been reviewed to determine the toxic nature of test chemical on the mortality of fish. The studies are as mentioned below:  

A 96 h study was conducted for test chemical on Danio rerio, at the test concentrations of 5mg/L, & 7.5 mg/L. In5mg/L, & 7.5 mg/L, the fishes were found to be freely swimming in the bowl aquaria. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 50 mg & 75 mg of the test substance in 10 liters deionized water with continuous stirring for achieving test concentrations of 5 mg/L and 7.5 mg/L, respectively. No mortalities were found in the control aquaria. Hence the LC50 value was considered to be > 7.5 mg/L and the LC0 value was determine to be 7.5 mg/l. Thus considering CLP Criteria for aquatic classification of the substance, it is concluded that test chemical exhibits short term toxicity to fish (Danio rerio) and classified in the chronic 2 classification category.

 

First study was supported by the second experimental study for test chemical. Short term toxicity of test chemical was studied in fathead minnows (Pimephales promelas) fish. Before testing chemical was analytically monitorized by gas liquid chromatography. Stock solution of test chemical (42.3 mg/l) were prepared by blending. 29 days old fishes were used for the study under the flow-through system. The 96 hrs Lethal concentration (LC50) and effective concentration (EC50) value of test chemical on fish Pimephales promelas was observed at dose concentration of 5.09 mg/l and 4.71 mg/l respectively. Thus considering CLP Criteria for aquatic classification of the substance, it is concluded that tes test chemical exhibits short term toxicity to fish (Pimephales promelas) and classified in the chronic 2 classification category.

 

Based on the above studies, it was conclude that the chemical was toxic and can be consider to be classified in aquatic chronic 2 category as per the CLP classification criteria. But as the chemical was readily biodegradable in water, thus on that basis chemical consider to be nontoxic and not classified as per the CLP classification criteria.