2-hydroxy-1-(4-(4-(2-hydroxy-2-methylpropionyl)benzyl)phenyl)-2-methylpropan-1-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

Study period:

2002-11-11 to 2003-01-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: duplicate sample of just before test start , after 48 h and 96 h the test media of all test concentrations and the control
- Sampling method: in the WAFs of the loading rates of 10 and 22 mg/L, all test fish were dead already after 24 and 72 hours test duration, samples in this concentrations were taken after these time periods
- Sample storage conditions before analysis: without mixing of the test media, and were deep-frozen (at about -20 °C) immediately after sampling

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Prior to the start of the test, five individual mixtures with loading rates of 1.0, 2.2, 4.6, 10, and 22 mg/L were prepared by mixing 5.0, 11.0, 23.0, 50.4, and 110.1 mg of the test item into 5000 mL test water, respecfively. No auxiliary solvent or emulsifier was used. The test item was mixed into the test water as homogeneously as possible by ultrasonification for 15 minutes and intense stirring. The dispersions were stirred on a magnetic stirrer at room temperature in the dark for 96 hours to dissolve a maximal amount of the test item in test water. After the sfirring period of 96 hours, the dispersions were filtered through a membrane filter just before the start of the test. The filtrates of the dispersions with dissolved test item were tested on the test fish as WAFs. Additionally, a control was tested in parallel.
- Eluate: test water
- Differential loading: 1.0, 2.2, 4.6, 10, and 22 mg/L
- Controls: yes (test water)

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish (Brachydanio rerio)
- Source: commercial supplier
- Age at study initiation (mean and range, SD): more than 4 weeks
- Length at study initiation (length definition, mean, range and SD): 3.3 ± 0.2 cm based on 10 fishs
- Weight at study initiation (mean and range, SD): 0.38 ± 0.05 g based on 10 fish
- Feeding during test: none (fishs were fed until one day before the start of the test)
- Food type: none

ACCLIMATION
- Acclimation period: one week
- Acclimation conditions (same as test or not): same
- Type and amount of food: a commercial fish diet
- Health during acclimation (any mortality observed): yes

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

2.5 mmol/L (= 250 mg/L) as CaCO3

Test temperature:

21-23 °C during the test period

pH:

7.8 to 7.9

Dissolved oxygen:

7.7 mg/L or higher

Nominal and measured concentrations:

- loading rates (mg/L): 1.0; 2.2; 4.6; 10, 22
- mean measured concentration: not analyzed; not analyzed; 2.7, 4.6, 5.5 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquarium with 4 liters test medium
- Type (delete if not applicable): open
- Aeration: yes
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: The loading rate was lower than 1 g fish wet weight per liter test medium.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconsfituted water
- Total organic carbon: The test water was aerated prior to the preparation of the test media until oxygen saturafion was reached.
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4 : 1 (based on molarity)
- Intervals of water quality measurement: at the start of the test and once every day during the test in all test concentrations and in the control

OTHER TEST CONDITIONS
- Photoperiod: The test was performed in the dark since solutions of the test item are sensifive to light.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and visible abnormalities, 3.5 , 24, 48, 72, 96 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: The loading rates were based on the results of a range-finding test and on the results of preexperimentsto the solubility of the test item.
- Justification for using less concentrations than requested by guideline: the solubility of the test item
- Range finding study: yes

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3.5 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 2.7-4.6 mg/L Confidence interval

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

2.7 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

2.7 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

4.6 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.:

6.8 mg/L

Nominal / measured:

nominal

Conc. based on:

other: loading rate

Basis for effect:

mortality (fish)

Remarks on result:

other: 4.6-10 mg/L Confidence interval

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

4.6 mg/L

Nominal / measured:

nominal

Conc. based on:

other: loading rate

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

4.6 mg/L

Nominal / measured:

nominal

Conc. based on:

other: loading rate

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

other: loading rate

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: One fish at the loading rate of 2.2 mg/L showed abnormal behaviour. However, the observed symptoms are considered not to be caused by the test item since at the next higher loading rate of 4.6 mg/L, no symptoms were observed in any test fish. At the next higher loading rate of 10 mg/L (mean measured 4.6 mg/L), all test fish showed one or several visible abnormalities and died within 72 hours test durafion. At the highest loading rate of 22 mg/L (mean measured 5.5 mg/L), all fish were dead already after 24 hours test durafion.
- Mortality of control: all fish survived unfil the end of the test
- Other adverse effects control: no visible abnormalities were observed

Sublethal observations / clinical signs:

The analytically measured test item concentrafions in the analyzed test media samples (loading rates of 4.6, 10, and 22 mg/L) amounted to 3.4, 4.9, and 5.6 mg/L at the start of the test. At the end of the test, 57% of the inifially measured test item concentrafion was found at the loading rate of 4.6 mg/L. At the loading rate of 10 mg/L, 87% of the inifially measured value was found after 72 hours (all test fish were dead after 72 hours test durafion). At the highest loading rate of 22 mg/L, 97% of the inifially measured concentration was determined after 24 hours (all test fish were dead already after 24 hours test durafion). The mean measured test item concentrations (calculated as the average over all measurements per test concentration) varied in the range of 79 to 98% of the inifially measured values.

The biological results are related to the loading rates of the WAFs and to the mean measured test item concentrafions (sum of both isomers). In the control and at the loading rates up to and including 4.6 mg/L (mean measured 2.7 mg/L), all fish survived unfil the end of the test and no visible abnormalities were observed. The only excepfion was one test fish at the loading rate of 2.2 mg/L which showed abnormal behavior. However, the observed symptoms are considered not to be caused by the test item since at the next higher loading rate of 4.6 mg/L, no symptoms were observed in any test fish. At the next higher loading rate of 10 mg/L (mean measured 4.6 mg/L), all test fish showed one or several visible abnormalities and died within 72 hours test durafion. At the highest loading rate of 22 mg/L (mean measured 5.5 mg/L), all fish were dead already after 24 hours test durafion. Thus, the 96-hour NOEC (highest concentration tested without observable toxic effects after the exposure period of 96 hours) and the 96-hour LC0 of the test item was determined to be at the loading rate of 4.6 mg/L (mean measured 2.7 mg/L). The 96-hour LOEC (lowest concentrafion tested with observable toxic effects) was determined to be at the loading rate of 10 mg/L (mean measured 4.6 mg/L).

The 96-hour LC50 of the test item based on loading rates was calculated to be 6.8 mg/L with a 95% confidence interval from 4.6 to 10 mg/L. The 96-hour LC50 of the test item based on mean measured test item concentrafions was calculated to be 3.5 mg/L with a 95% confidence interval from 2.7 to 4.6 mg/L. The 96-hour LC 100 was determined at the loading rate of 10 mg/L (mean measured 4.6 mg/L). Thus, the concentrafion-effect relationship of the test item was rather steep. No remarkable observations were made concerning the appearance of the test media. The test media were clear solufions throughout the enfire test duration. At the loading rate of 10 mg/L, the test medium was turbid after 72 hours test durafion.

Validity criteria fulfilled:

yes

Conclusions:

The test substance is acutely toxic to fish.

Executive summary:

In this guideline (OECD 203) study conducted with GLP certification, the 96 hour LC50 of the test material (EC 444-860-9) was determined to be 3.5 mg/l (mean measured). The test was conducted on zebra fish under static conditions. The test result does not trigger classification and labelling of the test material under the criteria of the EU Classification, Labelling, & Packaging (CLP) regulation (1272/2008).

Description of key information

Study conducted to recognised testing guidelines with GLP certification.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

3.5 mg/L

Additional information

The acute toxicity of the test substance to zebrafish (Danio rerio) under static conditions was determined according to OECD guideline 203 (RCC Ltd., 2003). Since the test item contains two different isomers, five WAFs (water accommodated fractions) with loading rates of 1.0, 2.2, 4.6, 10, and 22 mg/L were used. The test concentrations were analytically verified at the beginning and at the end of the test period.

In this GLP study the test substance was shown to be acutely toxic to fish. An LC50 (96 h) of 3.5 mg/L mean measured (= 6.8 mg/L loading rate) was determined. The NOEC was determined to be 2.7 mg/L mean measured (= 4.6 mg/L loading rate) after 96 h.