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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Link to relevant study record(s)

Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20/01/2015 - 27/01/2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study according to international guideline (OECD guideline 202) under GLP. No deviations from guideline reported.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
Principles of method if other than guideline:
Not relevant
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
spearmint oil
Mentha cardiaca, ext.
EC 294-809-8
CAS 91770-24-0
Oil of spearmint - Chinese type
Oil of spearmint - Scotch variety

Details on properties of test surrogate or analogue material (migrated information):
Not relevant
Analytical monitoring:
yes
Details on sampling:
- Concentrations: nominal water accommodated fractions (WAF) of 1.0, 2.2, 4.6, 10, 22 and 46 mg/l
- Sampling method: Samples for possible analysis were taken from all test concentrations and the control according to at t=0 h and t=48 h. 50 ml samples were taken from the approximate centre of the test vessels. At the end of the exposure period, the replicates were not pooled at each concentration before sampling.
- Sample storage conditions before analysis: Samples were stored in a refrigerator until analysis. Additionally, reserve samples of 50 ml were taken for possible analysis. If not used, these samples were stored in a freezer for a maximum of three months.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water Accommodated Fractions (WAFs) were prepared at individual loading rates ranging from 1.0 to 100 mg/l. Exact amounts of test substance were added to test medium and magnetically stirred for 2 days in closed vessels with minimal headspace. Thereafter, all solutions were left to stabilise overnight. Subsequently, the WAFs were collected by siphoning the water phase and distributed over the test vessels.
- Controls: yes, blanks
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna
- Strain: Straus, 1820, at least third generation, obtained by acyclical parthenogenesis under specified breeding conditions
- Source: In-house laboratory culture with a known history
- Age at study initiation (mean and range, SD): less than 24 hours old
- Method of breeding: Each batch was started with newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of Elendt M7 medium in an all-glass culture vessel. Cultures are 4 weeks old as a maximum. After 7 days of cultivation half of the medium was replaced twice a week. Cultures were held at 18-22°C and fed daily with a suspension of fresh water algae.
- Feeding during test: No
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
No
Hardness:
180 mg/l CaCO3
Test temperature:
19 - 20ºC
pH:
7.5 - 9.4 (no change exceeding 1.5 pH units during test)
Dissolved oxygen:
7.7 - 8.0 mg/l
Salinity:
Not relevant
Nominal and measured concentrations:
Nominal WAF: control, 1.0, 2.2, 4.6, 10, 22 and 46 mg/l
Measured (TOC, t=0h): 0.01526, 4.817, 2.411, 3.684, 15.84, 17.59, 28.49 mg/l
Measured (TOC, t=72h): 0.09637, 5.881, 2.439, 3.465, 15.33, 17.13, 28.41 mg/l
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: Glass, 60 ml, minimum headspace, closed airtight
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water purified by reverse osmosis
- Medium composition:
Macro salts: CaCl2.2H2O 211.5 mg/l
MgSO4.7H2O 88.8 mg/l
NaHCO3 46.7 mg/l
KCl 4.2 mg/l
Medium M7: trace elements, macro nutrients and vitamins are added to freshly prepared ISO medium to reach the following concentrations:
Trace elements: B 0.125 mg/l
Fe 0.05 mg/l
Mn 0.025 mg/l
Li, Rb and Sr 0.0125 mg/l
Mo 0.0063 mg/l
Br 0.0025 mg/l
Cu 0.0016 mg/l
Zn 0.0063 mg/l
Co and I 0.0025 mg/l
Se 0.0010 mg/l
V 0.0003 mg/l
Na2EDTA.2H2O 2.5 mg/l
Macro nutrients: Na2SiO3. 9H2O 10.0 mg/l
NaNO3 0.27 mg/l
KH2PO4 0.14 mg/l
K2HPO4 0.18 mg/l
Vitamins: Thiamine 75.0 µg/l
B12 1.0 µg/l
Biotin 0.75 µg/l

- Culture medium different from test medium: No
- Intervals of water quality measurement: pH and dissolved oxygen are measured at the beginning and at the end of the test, for all concentrations and the control. Temperature is monitored continually.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16h light / 8h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobility recorded at 24 hours and at 48 hours.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
- Range finding study
- Test concentrations: 1, 10 and 100 mg/l
- Results used to determine the conditions for the definitive study: yes
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Duration:
24 h
Dose descriptor:
EL50
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CL: 1.4 - 24 mg/l
Key result
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
3.1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CL: 2.5 - 3.6 mg/l
Details on results:
- Mortality of control: 0%
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- EC50/LC50: The 24h-EC50 was 0.93 mg/l with a 95% confidence interval between 0.79 and 1.0 mg/l. The 48h-EC50 was 0.47 mg/l with a 95% confidence interval between 0.42 and 0.52 mg/l.
Reported statistics and error estimates:
The 24 and 48h-EL50-value were calculated from the weibits of the percentages of affected daphnids and the logarithms of the corresponding loading rates using the maximum likelihood estimation method (Weibull analysis).

ToxRat Professional v 2.10.05 (ToxRat Solutions® GmbH, Germany) was used to perform the analyses.

Number of introduced daphnids and incidence of immobility in the second full test

Time (h)

Replicate

Spearmint Essential oil, ex Mentha cardiaca

WAF prepared at (mg/l)

Control

1.0

2.2

4.6

10

22

46

0

A

5

5

5

5

5

5

5

B

5

5

5

5

5

5

5

C

5

5

5

5

5

5

5

D

5

5

5

5

5

5

5

Total introduced

20

20

20

20

20

20

20

 

24

A

0

0

0

3

3

5 (1)

5

B

0

0

0

1

0

5

5

C

0

0

0

3

1

5

5

D

0

0

0

0

1

5

5

Total immobilised

0

0

0

7

5

20

20

Effect %

0

0

0

35

25

100

100

 

 

 

 

 

 

 

48

A

0

0

1

5

4

5

5

B

0

0

2

4

5

5

5

C

0

0

0

5

5

5

5

D

0

0

1

5

5

5

5

Total immobilised

0

0

4

19

19

20

20

Effect %

0

0

20

95

95

100

100

( ) between brackets: number of daphnia observed trapped at the surface of the test solutions. This organism was reimmersed into the respective solutions before recording of mobility.

pH and oxygen concentrations during the second full test

Spearmint Essential oil, ex Mentha cardiaca

WAF prepared at (mg/l)

Start (t=0 h)

End (t=48 h)

pH

O2

pH

O2

Control

9.4

7.9

8.5

8.0

1.0

9.1

7.9

8.1

7.9

2.2

9.0

7.9

7.6

7.8

4.6

9.0

7.9

7.7

7.8

10

9.1

7.8

7.6

7.7

22

9.0

7.9

7.5

7.7

46

8.9

7.8

7.6

7.7

Validity criteria fulfilled:
yes
Remarks:
Mortality in control < 10%, dissolved oxygen > 3 mg/l
Conclusions:
The acute toxicity (48h-EL50) of spearmint oil towards Daphnia magna is 3.1 mg/l.
Executive summary:

The acute toxicity of spearmint oil towards Daphnia magna was investigated according to OECD guideline 202 under GLP. Daphnia were exposed to nominal WAFs of 1.0, 2.2, 4.6, 10, 22 and 46 mg/l for 48 hours and observed. Based on nominal concentrations, the 24h-EL50 and 48h-EL50 were found to be 10 and 3.1 mg/l respectively.

Description of key information

48h-EL50: 3.1 mg/l

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
3.1 mg/L

Additional information

The acute toxicity of spearmint oil towards Daphnia magna was investigated according to OECD guideline 202 under GLP. Daphnia were exposed to nominal WAFs of 1.0, 2.2, 4.6, 10, 22 and 46 mg/l for 48 hours and observed. Based on nominal concentrations, the 24h-EL50 and 48h-EL50 were found to be 10 and 3.1 mg/l respectively.