Star anise, Illicium verum, ext.

Toxicity to microorganisms

Administrative data

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation))

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

Details on sampling:

Test substance information

Test item: Essential Oil of Star Anise Oil (Illicium verum)
Common name of the active ingredient (a.i.) UVCB substance (1)
Received on: 17/May/2018
Bioagri code: QUIM-0014/18
Analyzed Concentration of a.i. (Bioagri): 96.32% (w/w)
Analysis Certificate (CA): CA TH – 0461/17 R1
Batch number: 18-02-10-1 (1)
Test Substance sent by: Jinxi County Purong Essences Mfg Co., Ltd (1)
Manufacturer: Jinxi County Purong Essences Mfg Co., Ltd (1)
Address: JiangXi Jinxi County Industrial Park Area C (1)
Physical state: Light yellow clarified liquid or crystal (1)
Manufacture Date: 10/Feb/2018 (1)
Validity Date: 09/Aug/2019 (1)
Source: (1) Information provided by the sponsor.

Test solutions

Vehicle:

no

Details on test solutions:

Water Milli-Q
Sodium 1-heptanesulfonate 99%
Methanol 99.9%
Bacteriological Peptone
Meat Extract
Urea 99%
Sodium Chloride 99%
Calcium chloride 98%
Magnesium Sulfate 98%
Potassium phosphate 98%
Deionized water
Copper sulphate penta hydrate 98.22%
Water without chlorine

Test organisms

Test organisms (species):

activated sludge of a predominantly industrial sewage

Details on inoculum:

All chemicals used in the test were “analytical grade” or “for use in microbiology”.
Composition of nutrient solution (synthetic sewage):

Peptone 16.0 g
Beef extract 11.0 g
Urea 3.0 g
NaCl 0.7 g
CaCl2 *2H2O 0.4 g
MgSO4 *7H2O 0.2 g
K2HPO4 2.8 g
Deionised water 1000 mL

The pH of the solutions was 7.52. The medium was prepared and stored in the dark at a temperature of 4°C.

Study design

Water media type:

freshwater

Total exposure duration:

3 h

Test conditions

Test temperature:

20±2℃

pH:

7.52

Nominal and measured concentrations:

One replicate/treatment (positive control)
One replicate for concentration (10 mg L-1)
One replicate for concentration (100 mg L-1)
Three replicates for concentration (1000 mg L-1)

Details on test conditions:

The study was performed in a closed system with aeration. In the control vessels, 16 mL of nutrient solution was mixed with 234 mL of water. The positive control and the test vessels were prepared by adding the appropriate amount of positive control solution respectively test item into the test vessel, adding 16 mL nutrient solution and water to give 250 mL. Then, 250 mL inoculum was added in ten minutes intervals. After addition of the inoculums, the flasks were closed immediately and the mixtures were stirred with aeration.
After three hours the contents were poured into a tube of approximately 50 mL and the respiration rate was determined by measuring the O2 concentration over a ten minutes period. The following vials were measured equally at ten minutes intervals.

Reference substance (positive control):

yes

Remarks:

copper sulfate pent hydrate

Results and discussion

Effect concentrations

Key result

Duration:

3 h

Remarks on result:

other: there were no statistically significant toxic effects at the highest concentration (1000 mg L-1)

Details on results:

The test substance was solubilized in water because of its solubility. As there were no statistically significant toxic effects at the highest concentration (1000 mg L-1), no further testing was required at higher or lower concentrations as outlined in OECD 209, 2010.

Results with reference substance (positive control):

Variation in O2-Consumption of the Controls

Average oxygen uptake rate was 66.50 mg L-1 h. The variation of OD consumption in the controls was 15.71% which are far below the recommended upper limit of 30%.

Any other information on results incl. tables

Table 6. O₂-Consumption, Inhibition - test substance.

 

Test Concentration	pH	Consumption (mg L^-1h)	Inhibition (%)	Variation in DO consumption of controls (%)
Control C1	7.65	56.05	-	15.71
F1 – 1%	8.01	66.50	0.00
F2 – 10%	7.83	70.30	0.00
F3 – 100%	8.00	66.50	0.00
F3 – 100%	8.00	61.75	0.00
F3 – 100%	7.89	67.45	0.00
Abiotic control	7.24	0.00	-
Control C2	8.01	66.50	-

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

As there were no statistically significant toxic effects at the highest concentration (1000 mg L-1), no further testing was required at higher or lower concentrations as outlined in OECD 209, 2010.