[(3aS*,4R*,5S*,7R*,7aS*)-4,7-methano-2,3,3a,4,5,6,7,7a-octahydro-1H-inden-5-yl]methyl acetate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures"

Version / remarks:

December 15, 2000

Qualifier:

according to guideline

Guideline:

other: SANCO/3029/99 rev.4 11/07/00: Residues: Guidance for generating and reporting methods of analysis in support of preregistration data requirements for Annex II (part A; Section 4) and Annex III (part A; Section 5) of directive 91/414

Version / remarks:

rev. 4 11/07/00

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Nominal test concetrations of 0.85, 1.9, 4.1, 9.1 and 20 mg/L and the control in fresh media at 0 and 24 hours and aged media after 24 and 48 hours at test media renewal
- Sample storage conditions before analysis: Samples were extracted with cyclohexane immediately after sampling, and stored in a refrigerator at 4 ± 4 ºC, and protected from light until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test media of the nominal 20 mg test item/L solution was prepared by dissolving 20 and 20.4 mg test item into 1000 and 1020 mL test water by intense stirring for 24 hours. Adequate volumes of this test medium were diluted with test water to prepare the test media of the other desired test concentrations. The test media were prepared just before introduction of the daphnids at the start of the test and test medium renewal on Day 1.
- Controls: Reconstituted water (Elendt M4 media)
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): No

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain/clone: Clone 5
- Source: Bred at test facility
- Age of parental stock (mean and range, SD): Not reported, but test animals not first brood progeny
- Feeding during test: None

ACCLIMATION
- Acclimation period: None
- Acclimation conditions (same as test or not): Culture media conditions same as test conditions

BREEDING CONDITIONS
- Type and amount of food: Breeding cultures fed green algae (Desmodesmus subspicatus), amount not reported
- Feeding frequency: Stock cultures fed on all working days

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

Media renewal after 24 hours

Hardness:

2.5 mmol/L, equivalent to 150 mg/L, as CaCO3

Test temperature:

Test start: 19.8 to 20.4°C
Aged media at 24 hours: 20.0 to 20.1°C
Fresh media at 24 hours: 20.0 to 20.2°C
Aged media at 48 hours: 19.9 to 20.0°C

pH:

Test start: 8.0 to 8.3
Aged media at 24 hours: 7.8 to 7.9
Fresh media at 24 hours: 7.9
Aged media at 48 hours: 7.9 to 8.0

Dissolved oxygen:

Test start: 8.7 to 8.8 mg/L
Aged media at 24 hours: 8.5 to 8.7 mg/L
Fresh media at 24 hours: 8.8 mg/L
Aged media at 48 hours: 8.4 to 8.5 mg/L

Conductivity:

< 5 μS/cm

Nominal and measured concentrations:

Nominal: 0.85, 1.9, 4.1, 9.1 and 20 mg test item/L
Mean measured: 0.655*, 1.58, 3.40, 7.50 and 17.3 mg test item/L
*For values

Details on test conditions:

TEST SYSTEM
- Test vessel: Beaker
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: Glass, 110 mL, containing maximum amount of test medium possible, with the remaining headspace reduced to the technically possible minimum, with conical glass stopper.
- Renewal rate of test solution (frequency/flow rate): Media renewed after 24 hours
- 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: Analytical grade salts and additives added to deionised water at the following nominal concentrations;
Main compounds; CaCl2·2 H2O = 293.80 mg/L, MgSO4·7 H2O = 123.30 mg/L, KCl = 5.80 mg/L, NaHCO3 = 64.80 mg/L, Na2SiO3·9 H2O = 10.00 mg/L, NaNO3 = 0.27 mg/L, KH2PO4 = 0.14 mg/L, K2HPO4 = 0.18 mg/L
Trace elements: H3BO3 = 2860 μg/L, MnCl2·4 H2O = 361 μg/L, LiCl = 306 μg/L, RbCl = 71 μg/L, SrCl2·6 H2O = 152 μg/L, NaBr = 16 μg/L, Na2MoO4·2 H2O = 63 μg/L, CuCl2·2 H2O = 17 μg/L, ZnCl2 = 13 μg/L, CoCl2·6 H2O = 10 μg/L, KI = 3.3 μg/L, Na2SeO3 = 2.2 μg/L, NH4VO3 = 0.6 μg/L, Na2EDTA·2 H2O = 2.5 mg/L, FeSO4·7 H2O = 1.0 mg/L
Vitamins: Thiamin HCl = 75.0 μg/L, Cyanocobalamin (B12) = 1.0 μg/L, Biotin (B6) = 0.75 μg/L
- Metals: None reported except test media components
- Alkalinity: 0.9 mmol/L
- Ca/mg ratio: Not reported
- Conductivity: < 5 μS/cm
- Culture medium different from test medium: No
- Intervals of water quality measurement: Water temperature, pH and dissolved oxygen concentrations determined at test start, in aged and fresh media at 24 hours and test end.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light : 8 h dark
- Light intensity: 580 - 640 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mobility after 24 and 48 hours

RANGE-FINDING STUDY
- Results used to determine the conditions for the definitive study: Yes

Reference substance (positive control):

yes

Remarks:

Potassium dichromate tested at least twice per year

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

6.21 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CI: 4.98 - 7.76 mg/L

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

3.1 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CI: 2.17 - 4.41 mg/L

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.655 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

7.33 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CI: 5.81 - 9.24 mg/L

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

3.46 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CI: 2.39 - 5.02 mg/L

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

0.655 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: 0 after 48 hours
- Other adverse effects control: None
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: 24 hrs EC50 must be between 0.6 and 2.1 mg potassium dichromate/L
- EC50: 24 hrs EC50: 0.959 mg/L; 48 hrs EC50: 0.765 mg/L, 95% CI: 0.650 - 0.888 mg/L;

Reported statistics and error estimates:

The 24-hour and 48-hour EC50, EC10 and the 95 % confidence limits were calculated by logit analysis. The NOEC and LOEC after 24 and 48 hours were determined directly from the raw data. All analysis performed using ToxRat Professional, Version 3.2.1.

Summary of Analytical Results

Nominal Test Concentration    (mg test item/L)	Fresh test media (0 h)			Aged test media (24 h)			Mean measured concentration
	% of nominal ^1	RSD (%)	n	% of nominal^1	RSD (%)	n	mg test item/L	n
Control	n.a.	n.a.	4	n.a.	n.a.	4	n.a.	8
0.85	91	9	4	77	12	4	0.655	8
1.9	88	12	4	79	13	4	1.58	8
4.1	87	11	4	78	11	4	3.40	8
9.1	86	10	4	79	12	4	7.50	8
20	91	5	4	82	10	4	17.3	8

1: mean value of all measured samples per treatment group

italics: Values below the LOQ. For values <LOQ, half the LOQ was used for calculation measured concentrations according to OECD Guidance Document No. 23

RSD: relative standard deviation per treatment group

n: number of samples analysed

n.a.: not applicable

Influence of the test itme on the mobility of Daphnia magna

Mean measured     (mg test item/L)	No. of Daphnia tested	No. of immobilised Daphnia after		% of immobilised Daphnia after
		24h	48h	24h	48h
Control	20	0	0	0	0
0.655	20	0	0	0	0
1.58	20	1	1	5	5
3.4	20	2	2	10	10
7.5	20	8	12	40	60
17.3	20	20	20	100	100

Influence of test item on the mobility of Daphnia magna

Endpoint (mg test item/L)	24 hours	48 hours
EC50	7.33	6.21
95 % CI	5.81 - 9.24	4.98 - 7.76
EC20	4.57	4.00
95 % CI	3.41 - 6.10	3.02 - 5.30
EC10	3.46	3.10
95 % CI	2.39 - 5.02	2.17 - 4.41
NOEC	0.655	0.655
LOEC	1.580	1.580

Values refer to mean measured test concentrations

CI: Confidence Interval

NOEC and LOEC determined directly from the raw data

Validity criteria fulfilled:

yes

Conclusions:

Based on mean measured concentrations, the 48-hour EC50 for the test item to Daphnia magna was 6.21 mg test item /L, with 95 % confidence interval of 4.98 - 7.76 mg test item/L. The 48-hour NOEC was 0.655 mg test item/L.

Executive summary:

The acute toxicity of the test item to Daphnia magna was determined in a static renewal test following OECD guideline 202, with media renewal after 24 hours. Daphnia were exposed to nominal concentrations of 0.85, 1.9, 4.1, 9.1 and 20 mg test item/L, equivalent to mean measured concentrations of 0.665, 1.58, 3.40, 7.50 and 17.3 mg test item/L, alongside a control. Based on mean measured concentrations, the 48-hour EC50 was 6.21 mg test item/L, with 95 % confidence interval of 4.98 to 7.76 mg test item/L. The 48-hour NOEC was 0.655 mg test item/L.

 

The study is a GLP compliant, guideline experimental study available as an unpublished study report. There are no restrictions and the study is fully adequate for assessment.

Description of key information

The 48-hour EC50 of the test item to Daphnia magna was determined to be 6.21 mg test item/L, with 95 % confidence interval of 4.98 to 7.76 mg test item/L, based on mean measured concentrations. The 48 -hour NOEC was 0.655 mg test item/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

6.21 mg/L

Additional information

The acute toxicity of the test item to Daphnia magna was determined in a static renewal test following OECD guideline 202, with media renewal after 24 hours. Daphnia were exposed to nominal concentrations of 0.85, 1.9, 4.1, 9.1 and 20 mg test item/L, equivalent to mean measured concentrations of 0.665, 1.58, 3.40, 7.50 and 17.3 mg test item/L, alongside a control (2017). The study is a GLP compliant, guideline experimental study available as an unpublished study report. There are no restrictions with the study is fully adequate for assessment..