Methyl 2-[(1,5-dihydro-3-methyl-5-oxo-1-phenyl-4H-pyrazol-4-ylidene)ethylidene]-1,3,3-trimethylindoline-5-carboxylate

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Control: after 24 hours
- Test concentrations: at 0, 24 and 72 hours in the freshly prepared media and at 24, 48 and 96 hours in the aged media after 24 hours of exposure

Vehicle:

no

Details on test solutions:

Pre-treatment of test item and preparation of test item concentrations
- direct weighing was prepared each day to give the desired test concentration
- 500 mg of the test item were added to 5 litres of dilution water
- treated for 1 h in an ultrasonic bath and stirred for 24 h on a magnetic stirrer
- Undissolved particles of the test item were removed by filtration using a folded filter (pore size 7 - 12 μm)
- pH ranged between 7.3 and 7.5

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

- Name: Zebra fish (Danio rerio)
- Source: Interaquaristik.de Shop (Germany)
- Date supplied: 2016-08-24
- Acclimatisation: Stock held since 2016-08-24 and acclimatised to the test conditions since then
- Temperature: 20 - 24 °C
- Dissolved oxygen: > 5 mg/L
- Feeding: Commercial fish food, daily; feeding discontinued 24 h prior to test start.
- Mortalities during acclimatisation period: < 5 %
- Medication: none
- Mean standard length (n = 14): 4.62 cm (Standard Deviation = 0.21 cm)

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

14 °dH

Test temperature:

21.2 - 22.5 °C

pH:

7.3 - 7.8

Dissolved oxygen:

7.4 - 9.0 mg/L

Nominal and measured concentrations:

100 mg/L nominal;
0.209 mg/L after 0 h of exposure (measured), 0.224 after 96 h of exposure (measured)

Details on test conditions:

Exposure conditions
- Test vessels: glass aquaria holding 5 L of test media covered by glass plates
- Experimental design: 1 test concentration plus 1 control, 7 fish per test concentration, no feeding during the exposure period
- Method of initiation: fish were placed in prepared media
- Loading: 1.42 g body weight (wet weight) per litre
- Photoperiod: 16 h light : 8 h dark
- Temperature: 21.2 to 22.5°C
- Aeration: gentle aeration via narrow glass tubes
- Test item concentration/s: 100 mg/L
- Method of administration: direct weighing
- Medium renewal: none / daily / flow-through system
- Duration of exposure: 96 hours
- Criteria of effects: absence of response to physical stimulation; in addition to observations on mortality at 2, 24, 48, 72 and 96 hours, type and incidence of sub-lethal effects compared with control fish were observed

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.216 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Effective concentrations ranged from 0.21 % to 0.33 % of nominal values in the freshly prepared medium/media and from 0.13 % to 0.22 % of nominal values in the medium/media after 24 hours of exposure according to the semi-static test conditions.

Validity criteria fulfilled:

yes

Remarks:

mortality in the controls < 10% by the end of the test; dissolved oxygen concentration > 60% of the air-saturation value throughout the exposure period; pH did not vary by more than 1 unit

Conclusions:

No toxic effects of Macrolex Orange R against fish were observed at a limit test concentration of nominally 100 mg/L under exposure conditions. A 96h-LC50 > 0.216 mg/L has been determined. All results are expressed in terms of geometric mean measured concentrations.

Executive summary:

The study was conducted in accordance with Council Regulation (EC) No 440/2008, Method C.1 'Acute toxicity for Fish' (2008) which is in most parts equivalent to the OECD Guideline for Testing of Chemicals No. 203 'Fish, Acute Toxicity Test' (1992) to assess the acute toxicity of Macrolex Orange R to Danio rerio under semi-static conditions.

An acute daphnia toxicity test and an algal growth inhibition test preceded the fish test. They provided information about the concentration which was used in the fish test. Because of the anticipated low toxicity of the test item, fish were exposed in the main test to a limit test concentration of nominally 100 mg/L for a period of 96 hours.

A group of seven fish of the recommended size was exposed to a limit test concentration of nominally 100 mg/L of Macrolex Orange R dissolved in dilution water. Auxiliaries used to prepare the test media were an ultrasonic bath, a magnetic stirrer and a folded filter. The test solution was changed every 24 h. During the test a temperature range of 20 - 24 °C was maintained in the test vessels, with a maximum temperature fluctuation of +/- 1.3 °C in the test. The temperature, the pH and the oxygen values were measured every day in the freshly prepared media and in the media after 24 hours of exposure.

Observations were made on the number of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure. No toxic effects against fish were observed at the limit of water solubility under exposure conditions. A 96h-LC50 > 0.216 mg/L (geometric mean measured concentration) has been determined.

All results are expressed in terms of nominal concentrations and geometric mean measured concentrations. Effective concentrations ranged from 0.21 % to 0.33 % of nominal values in the freshly prepared medium/media and from 0.13 % to 0.22 % of nominal values in the medium/media after 24 hours of exposure according to the semi-static test conditions.

Description of key information

The study was conducted in accordance with Council Regulation (EC) No 440/2008, Method C.1 'Acute toxicity for Fish' (2008) which is in most parts equivalent to the OECD Guideline for Testing of Chemicals No. 203 'Fish, Acute Toxicity Test' (1992) to assess the acute toxicity of Macrolex Orange R to Danio rerio under semi-static conditions.

An acute daphnia toxicity test and an algal growth inhibition test preceded the fish test. They provided information about the concentration which was used in the fish test. Because of the anticipated low toxicity of the test item, fish were exposed in the main test to a limit test concentration of nominally 100 mg/L for a period of 96 hours.

A group of seven fish of the recommended size was exposed to a limit test concentration of nominally 100 mg/L of Macrolex Orange R dissolved in dilution water. The test solution was changed every 24 h. No toxic effects against fish were observed at the limit of water solubility under exposure conditions. A 96h-LC50 > 0.216 mg/L has been determined.

All results are expressed in terms of nominal concentrations and geometric mean measured concentrations. Effective concentrations ranged from 0.21 % to 0.33 % of nominal values in the freshly prepared medium/media and from 0.13 % to 0.22 % of nominal values in the medium/media after 24 hours of exposure according to the semi-static test conditions.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.216 mg/L

Additional information

Should read: LC50 > 0.216 mg/L