A mixture mainly based on: 2,3-dihydro-6-(2-hydroxy-2-methyl-1-oxopropyl)-1,1,3-trimethyl-3-[4-(2-hydroxy-2-methyl-1-oxopropyl)phenyl]-1H-indene; 2,3-dihydro-5-(2-hydroxy-2-methyl-1-oxopropyl)-1,1,3-trimethyl-3-[4-(2-hydroxy-2-methyl-1-oxopropyl)phenyl]-1H-indene

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:

1988

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)

Deviations:

not specified

GLP compliance:

yes

Specific details on test material used for the study:

A sample of the test substance Article A was received from Fratelli Lamberti SPA, 21041 Albizzate, Italy, on 11 January 1988.

The test substance was a vitreous, high viscosity solid and the Sponsor's reference was Article A; it was assigned the Brixham code R024.

The test substance was stored in the dark at ambient temperature as the sponsor had stated that the test substance decomposed in sunlight.

Details on sampling:

The concentrations of Artiele A in the test solutions was measured at the start of the test and after 24 hr exposure and the freshly prepared solutions at 72 hr and at the end of the test (96 hr).

Vehicle:

yes

Remarks:

Acetone

Details on test solutions:

The test solutions were prepared by dissolving the required amount of Article A in 2 mI of acetone; this was then shaken before being added to 10 litres of dilution water and a second volume of 10 l of water added to ensure complete mixing.

The test solutions were replaced every 24 hr throughout the study.

The following nominaI exposure concentrations were employed:

10 mg Article A/litre, solvent control and dilution water control

The 10 mg/l solutions were slightly cloudy white in appearance.

AlI test vessels were set up in duplicate. Each vessel contained 100 µl/l of acetone except the dilution water control.

Dilution water:
The dilution water used in the study was the Brixham town freshwater supply which is delivered to the Laboratory via a holding tank (approximate residence time 24 hr). The water is dechlorinated by treatment with sodium thiosulphate and then passed through a carbon filter. It is equilibrated to the test temperature be fare use.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

The test organism was the rainbow trout (Salmo gairdneri), a sensitive freshwater fish.

The fish were obtained from Upwey Trout Farm, Bayard House, Upwey, Weymouth, Dorset, on 12 January 1988.

The fish had received a Halachite Green prophylatic treatment approximately four weeks before the start of the study.

Prior to the start of the study the fish were acclimated to the test temperature (15 +/- 1°C) for at least seven days .

During the holding and acclimation the fish were fed appropriate amounts of a commerciaI fish food .

The fish tested ranged in weight from 1.06 to 3.12 g with a mean weight of 1.8 g . The range in length was 44 to 61 mm with a mean length of 51.9 mm (data as measured on the control fish at the end of the study).

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

250 mg CaCO3/L

Test temperature:

15 ± 1 °C

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal: 10 mg/L

Details on test conditions:

Test procedure:
The test vessels used in this study were of alI glass construction, rectangular in shape, with dimensions 400 x 275 x 275 mm and a nominaI capacity of 30 litres. Two vessels were used for the exposure eoncentration, solvent and dilution water controIs.
All test solutions were gently aerated by passing eompressed air through glass pipettes. The temperature was maintained at 15 +1-loC and the light regime was fluorescent lighting for 16 hr and darkness for 8 hr.
The fish were not fed throughout the period of the test.
At the start of the test 20 litres of solution were prepared in each of the duplicate test vessels. Ten fish were added to each vessel giving a fish loading of 0.9 g fish per litre of solution, based on 20 litres of solution and the mean weight of fish (3.3).
The test solutions were renewed every 24 hr throughout the period of the test.

Observation of effects:
Observations of dead fish were recorded at 24, 48, 72 and 96 hr during the test, and at the same time a record of any symptoms of toxicity was made.

Physical and chemical analysis:
Daily measurements were undertaken throughout the 96 hr period for pH, dissolved oxygen and temperature in the test vessels and for hardness, eonductivity and pH of the dilution water.
The concentrations of Artiele A in the test solutions was measured at the start of the test and after 24 hr exposure and the freshly prepared solutions at 72 hr and at the end of the test (96 hr). The analytical method is described in Appendix l.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 3.7 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Biological data:
No mortalities or syrnptoms of toxicity were recorded in any test vessel.
There were no mortalities at the nominaI 10 mg/l concentration of Article A during the 96 hr exposure; the 24, 48, 72 and 96 hr LC50 values were alI therefore greater than the mean measured test substance concentration value aehieved in this test of 3.7 mg/l of Article A.

Physical and chemical data:
The parameters monitored daily in each test vessel.
The dissolved oxygen levels ranged from 8.8 to 9.8 mg/l.
The pH values ranged from 7.4 to 7.9.
The temperatures ranged from 14.8 to 16.0·C.
The total hardness of the dilution water was in the range 48.3 - 51.7 mg/l CaC03 , the conductivity 130 pS/cm and pH 7.6 - 7.8.

The concentrations of the test substance determined in the test solutions. The mean measured value of Article A in the nominal 10 mg/l expo su re vessels was 3.7 mg/l with a standard deviation of 0.77 mg/l.

Sublethal observations / clinical signs:

COMMENTS ON TEST PROCEDURE:

An aerated semi-static test procedure was selected for the study as the Sponsor had stated Article A to be stable in water but to decompose in sunlight. The test solution was replaced every 24 hr to prevent any significant loss of Article A by photodegradation, this was confirmed by analysis of the test solutions.

The water solubility of Article A was quoted as 10-15 mg/l. However, within the test system a water solubility of only approximately 4.0 mg/l could be achieved even with the use of 100 µl/l of acetone to aid dispersion.

Validity criteria fulfilled:

yes

Conclusions:

Article A was not acutely toxic at the level of its solubility in the test system and the 24, 48, 72 and 96 hr LC50 values were found to be >3.7 mg/l Article A based on mean measured test substance concentrations.

Executive summary:

Article A was not acutely toxic at the level of its solubility in the test system and the 24, 48, 72 and 96 hr LC50 values were found to be >3.7 mg/l Article A based on mean measured test substance concentrations.

Description of key information

Study conducted to recognised testing guidelines with GLP certification.

Additional information

LC50 > 3.7 mg/L.

The substance does not meet the criteria for classification in accordance with the classification, labelling and packaging regulation (EC 1272/2008).