Star anise, Illicium verum, ext.

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

OECD 202 (2004)

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

(E)-anethole

EC Number:

224-052-0

EC Name:

(E)-anethole

Cas Number:

4180-23-8

Molecular formula:

C10H12O

IUPAC Name:

1-methoxy-4-prop-1-en-1-ylbenzene

Constituent 2

Reference substance name:

4-allylanisole

EC Number:

205-427-8

EC Name:

4-allylanisole

Cas Number:

140-67-0

Molecular formula:

C10H12O

IUPAC Name:

1-allyl-4-methoxybenzene

1

Reference substance name:

Caryophyllene

EC Number:

201-746-1

EC Name:

Caryophyllene

Cas Number:

87-44-5

Molecular formula:

C15H24

IUPAC Name:

4,11,11-trimethyl-8-methylenebicyclo[7.2.0]undec-4-ene

Constituent 3

Reference substance name:

2,6-dimethyl-6-(4-methyl-3-pentenyl)bicyclo[3.1.1]hept-2-ene

EC Number:

241-702-9

EC Name:

2,6-dimethyl-6-(4-methyl-3-pentenyl)bicyclo[3.1.1]hept-2-ene

Cas Number:

17699-05-7

Molecular formula:

C15H24

IUPAC Name:

2,6-dimethyl-6-(4-methylpent-3-en-1-yl)bicyclo[3.1.1]hept-2-ene

Test material form:

liquid

Sampling and analysis

Analytical monitoring:

not specified

Test solutions

Vehicle:

no

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: obtained from the CAROLINA BIOLOGICAL SUPPLY COMPANY - Burlington, North Carolina (USA) and maintained as a stock culture at BIOAGRI.
- Age of parental stock (mean and range, SD): ≤ 24 hours old
- Feeding: In each culture container 30 organisms were maintained daily. They were daily fed with algae suspension (Pseudokirchneriella subcapitata) with 107 cells.organisms-1. The water of each culture was renewed twice a week (Mondays and Thursdays).

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: Nearly 24 hours before the Acute Toxicity Test started, Daphnia with eggs were isolated in crystallizing dishes and fed with algae suspension

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

148.1 mg.L-1 CaCO3

Test temperature:

20.2 to 20.3 °C

pH:

7.7 to 7.8

Dissolved oxygen:

8.0 mg.L-1

Details on test conditions:

In the range-finding test the exposure conditions were:

a): Total number of organisms per each tested concentration or control: 10;
b): Number of replicates per each tested concentration and control: 2;
c): Number of organisms per test container: 5;
d): Volume: 15 mL of test solution per test container, 3 mL to each Daphnia;
e): The organisms were distributed randomly in the test container. Firstly, three organisms were placed in each test container and, finally, two organisms were added to the container;
f): Nominal used concentrations: control and 0.1, 1; 10 and 100 mg.L-1;
g): Temperature of the test solutions: 20.2 to 20.3°C;
h): Photoperiod: 16 hours of light and 8 hours of dark;
i): Parameters evaluated: immobility after 24 and 48 hours; and still animals were considered unable to swim for a period of 15 seconds of observation after mild agitation of the flasks.

In the definitive test the exposure conditions were:

a): Total number of organisms per each tested concentration or control: 20;
b): Number of replicates per each tested concentration and control: 4;
c): Number of organisms per test container: 5;
d): Volume: 15 mL of test solution per test container, 3 mL to each Daphnia;
e): The organisms were distributed randomly in the test container. Firstly, three organisms were placed in each test container and, finally, two organisms were added to the container;
f): Nominal used concentrations: control and 6.5, 13; 25; 50 and 100 mg.L-1;
g): Temperature of the test solutions: 20.0 to 20.3°C;
h): Photoperiod: 16 hours light and 8 hours dark;
i): Parameters evaluated: immobility after 24 and 48 hours; and still animals were considered unable to swim for a period of 15 seconds of observation after mild agitation of the flasks;
j): Analytical determination: content dissolved oxygen, pH of the test solutions and the test substance concentration were determined at the beginning of the test and after 48 hours.

Reference substance (positive control):

yes

Remarks:

potassium dichromate p.a.

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

29.96 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobility

Remarks on result:

other: 95% confidence limits = 24.71 to 36.34 mg.L-1

Remarks:

95% confidence limits = 24.71 to 36.34 mg.L-1

Effect concentrations 2

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

35.53 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobility

Remarks on result:

other: 95% confidence limits = 29.46 to 42.84 mg.L-1

Remarks:

95% confidence limits = 29.46 to 42.84 mg.L-1

Effect concentrations 3

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

6.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobility

Effect concentrations 4

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobility

Details on results:

In the range-finding test, the number of immobile organisms at 24 and 48 hours for each tested concentration and control shoes that in 48 hours of exposure, the lowest concentration that caused 100% immobilization to the organisms was 100 mg.L-1, and the highest concentration that caused 0% immobilization to the organism was 10 mg.L-1.

The 24 and 48-hours EC50 values of the product of Essential oil of Star Anise Oil (Illiciumverum) obtained from the leaves and fruits by steam distillation were 35.53 mg.L-1 (95% confidence limits = 29.46 to 42.84 mg.L-1) and 29.96 mg.L-1 (95% confidence limits = 24.71 to 36.34 mg.L-1), respectively.

In the definitive test the highest tested concentration that did not cause acute effect (immobility) to the organisms after 48 hours was 6.5 mg.L-1. The lowest concentration that caused 100% immobility was 100 mg.L-1.

The 24-hours EC50 value of the reference substance (potassium dichromate) was 1.06 mg.L-1 (95% confidence limits = 0.93 to 1.22 mg.L-1), showing the sensitivity of the test system (ISO 6341 / OECD 202, 2004).

The values of dissolved oxygen and pH of the test concentrations and control shows that the values of pH of the control and test solutions measured at 0 and 48 hours not deviate more than 1.5 units during the test.

Results with reference substance (positive control):

The 24-hours EC50 value of the reference substance (potassium dichromate) was 1.06 mg.L-1 (95% confidence limits = 0.93 to 1.22 mg.L-1), showing the sensitivity of the test system (ISO 6341 / OECD 202, 2004).

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 48-hours EC50 value of Essential oil of Star Anise Oil (Illiciumverum) obtained from the leaves and fruits by steam distillation was 29.96 mg.L-1 (95% confidence limits = 24.71 to 36.34 mg.L-1).

Executive summary:

This study was carried out in order to evaluate the acute toxicity ofEssential oil of Star Anise Oil (Illiciumverum) obtained from the leaves and fruits by steam distillationtoDaphnia magna.A range-finding test was carried out to determine the concentrations that caused 100% and 0% of acute toxic effect. Based on these results, a definitive test was carried out using the following nominal concentrations: 6.5; 13; 25; 50; and 100 mg.L^{-. Acontrol were also added. Twenty organisms were used per concentration and control distributed into four replicates. The definitive test was conducted in a controlled environment with temperature ranging from 20.0to20.3°C for 48 hours with}photoperiod of16 hours light and 8 hours of dark. The 24, and 48-hours EC₅₀values ofEssential oil of Star Anise Oil (Illiciumverum) obtained from the leaves and fruits by steam distillationwere 35.53 mg.L^-1and 29.96 mg.L^-1, respectively. The highest concentration that did not cause acute effect (immobility) to the organisms in 48 hours was 6.5 mg.L^-1, and the lowest one that caused 100% of immobilization was 100 mg.L^-1.Thestatistical method used was Trimmed Spearman-Karber.