Eugenol

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:

19 Nov 2012 to 23 Nov 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study in compliance with international recognised guidelines

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0 & 10 mg/L

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Directly dissolution within test medium
- Controls: Test medium
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Strain: Hamilton-Buchanan
- Source: Dehner Garten Center, 67433 Neustadt/W.
- Age at study initiation (mean and range, SD): sexually immature young fish
- Length at study initiation (length definition, mean, range and SD): 2±1 cm
- Feeding during test: None

ACCLIMATION
- Acclimation period: 14 days under test conditions
- Acclimation conditions (same as test or not): yes
- Type and amount of food: warm water fish food and daphnia, totalling to about 1-2% of body weight per day
- Feeding frequency: three times a day
- Health during acclimation (any mortality observed): mortality didn't surpass 5%.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None

Hardness:

1.08 mmol/L

Test temperature:

21.0-22.1°C

pH:

7.7-8.0

Dissolved oxygen:

7.9 - 9.2 mg/L

Salinity:

not applicable

Nominal and measured concentrations:

nominal: 0, 10, 18, 32, 56, 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquaria
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 12.5 L
- Aeration: accomplished with glass tubes, frequency of bubbles 1/second
- Renewal rate of test solution (frequency/flow rate): every 24±1 hour
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water
- Total organic carbon: 0.4 mg/L
- Pesticides and biocides: - Chloride: 14 mg/L
- Conductivity: 330 µS/cm

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod:
- Light intensity:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Range finding study: yes
- Test concentrations of range finding study: 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

24 h

Dose descriptor:

LC100

Effect conc.:

18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC100

Effect conc.:

18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC100

Effect conc.:

18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: none
- Mortality of control: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none
- Effect concentrations exceeding solubility of substance in test medium: no

Reported statistics and error estimates:

The estimation of the LC50 was accomplished using the software Origin.

Validity criteria fulfilled:

yes

Conclusions:

The following results for the test item Eugenol USP 906 were determined:
96h NOEC = 10 mg/L
96h LC50 = 13 mg/L

Executive summary:

The study was conducted under GLP and according to OECD Guideline 203. The study was conducted in a closed semi-static system, and the test substance was replenished every 24 hours on the basis that eugenol is volatile. Fish were exposed to the substance at a range of concentrations from 10 to 100 mg/L in freshwater. The test was performed with 7 fish per vessel and concentration. For each concentration, the percentage of mortality at 24, 48, 72, and 96 hours was recorded. The following OECD 203 validity criteria were met: mortality in the control did not exceed 10% at the end of the test; the dissolved oxygen concentration did not drop below 60% throughout the test; constant conditions (temperature and pH) were within specified deviations; and the concentration of the substance was at least 80% of the nominal concentration throughout the test. Under the conditions of the test, the 96 hour LC50 was 13 mg/L.

Description of key information

OECD guideline 203, GLP, key study, validity 1 
96h -LC50 = 13 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

EC50

Effect concentration:

13 mg/L

Additional information

One acute toxicity study was performed to assess the acute toxicity of eugenol to the freshwater fish species Danio rerio (LAUS GmbH, 2013). The study was conducted under GLP and according to OECD Guideline 203. The study was conducted in a closed semi-static system, and the test substance was replenished every 24 hours on the basis that eugenol is volatile. Fish were exposed to the substance at a range of concentrations from 10 to 100 mg/L in freshwater. The test was performed with 7 fish per vessel and concentration. For each concentration, the percentage of mortality at 24, 48, 72, and 96 hours was recorded. The following OECD 203 validity criteria were met: mortality in the control did not exceed 10% at the end of the test; the dissolved oxygen concentration did not drop below 60% throughout the test; constant conditions (temperature and pH) were within specified deviations; and the concentration of the substance was at least 80% of the nominal concentration throughout the test. Under the conditions of the test, the 96 hour LC50 was 13 mg/L.