[3R-(3α,3aβ,6α,7β,8aα)]-octahydro-6-methoxy-3,6,8,8-tetramethyl-1H-3a,7-methanoazulene

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:

10 October - 20 November 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

(1992)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

(2008)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, December 14, 2000

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Test concentrations were verified by chemical analysis. Water samples (3.0 mL from the approximate centre of the test vessels) were taken from the control and each exposure level at t = 0 and t = 72h (fresh solutions) and at t = 24 and 96h (old solutions) . Samples were stored in the freezer until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
Preparation of test solutions started with a loading rate of 100 mg/L applying a two-day period of magnetic stirring followed by an one-hour settlement period. The aqueous Water Soluble Fraction (WSF) was siphoned off and used as the highest test concentration. The lower test concentrations were prepared by subsequent dilutions of the WSF in test medium. The final test solutions were all clear and colourless.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp (Cyprinus carpio)
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands
- Age at study initiation (mean and range, SD): not reported
- Length at study initiation (final test): 2.1 ± 0.10 cm
- Weight at study initiation (final test): 0.14 ± 0.05 g
- Method of breeding: F1 from a single parent-pair bred in UV-treated water.
- Feeding during test: No feeding from 24 hours prior to the test and during the total test period.

HOLDING / ACCLIMATION
- Acclimation period: at least 12 days after delivery
- Acclimation conditions: same as test, i.e. adjusted ISO medium
- Type of food: pelleted fish food (300-500 µm)
- Feeding frequency: daily
- Health during acclimation: <5% mortality observed during the seven days prior to the start of the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

180 mg/L (as CaCO3)

Test temperature:

21 - 22°C

pH:

7.5 - 8.1

Dissolved oxygen:

6.7 - 9.2 mg O2/L

Nominal and measured concentrations:

- Nominal concentrations: 0 (control), 2.2, 4.6, 10, 22, 46 and 100% of the WSF prepared at 100 mg/L (based on range-finding test).
- Measured concentrations (geom. mean): n.d., 0.022, 0.081, 0.17, 0.35, 0.81 and 1.1 mg/L.

The measured concentration decreased to 2.8 – 48% of initial.

Details on test conditions:

TEST SYSTEM
- Test vessel: 3.2 litres, all-glass, air-tight
- Fill volume: 3 litres of test solution
- Aeration: no
- Renewal rate of test solution: 24 h
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.33 g fish/litre, i.e.7 fish per 3 litres of test medium

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Adjusted ISO medium with a hardness of 180 mg CaCO3 per litre and a pH of 7.7 ± 0.3

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours photoperiod daily

EFFECT PARAMETERS MEASURED
- Mortality (after 3, 24, 48, 72 and 96 hours following the start of exposure)
- Other effects: clinical effects (such as hypoactive swimming, loss of equilibrium or immobility) - daily

PHYSICOCHEMICAL PARAMETERS MEASURED
Dissolved oxygen content, pH and temperature were measured daily in all vessels with surviving fish, beginning at the start of the test (day 0).

Reference substance (positive control):

yes

Remarks:

pentachlorophenol

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.43 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL 0.29 - 1.0 mg/L

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: 100% at 0.46 mg/L
- LC50: 0.32 mg/L (January 2015)
- Other: effect (i.e. 50% mortality) reached already within 48 h of exposure

Sublethal observations / clinical signs:

  Table: Incidence of mortality and total mortality during the test

Average measured exposure concentration (mg/L)	Cumulative mortality					Total mortality (%)
	2.75h	24h	48h	72h	96h
0 (control)	0	0	0	0	0	0
0.022	0	0	0	0	0	0
0.081	0	0	0	0	0	0
0.17	0	0	0	0	0	0
0.35	0	0	0	0	2	29
0.81	0	0	5	6	7	100
1.1	0	7	7	7	7	100

 

Table: Clinical effects observed during the test 

Average measured exposure concentration (mg/L)	Time (h)	Specification of effects	Number of fish
0.35	72	Hypoactive swimming	1/7
		Hypoactive swimming and snapping at the surface	4/7
		Hypoactive swimming, loss of equilibrium, and snapping at the surface	2/7
	96	Snapping at the surface	4/5
		Hypoactive swimming and loss of equilibrium	1/5
0.81	24	Hypoactive swimming	7/7
	48	Hypoactive swimming and loss of equilibrium	2/2
	72	Hypoactive swimming and loss of equilibrium	1/1

 

Table: Average exposure concentrations vs. nominal concentrations  

Nominal Concentration (mg/L)*	Measured concentration (mg/L)
	t=0h (fresh)	T=24h (old)	t=72h (fresh)	T=96h (old)	Average
2.2	0.025	0.00072	0.0873	0.018	0.022
4.6	0.0949	0.020	0.234	0.060	0.081
10	0.140	0.047	0.443	0.142	0.17
22	0.317	0.117	1.04	0.256	0.35
46	0.738	0.287	2.00	0.676	0.81
100	1.57	0.750	n.a.	n.a.	1.1

* As % of the WSF prepared at 100 mg/L

Validity criteria fulfilled:

yes

Remarks:

See 'Any other information on materials and methods incl. tables'

Conclusions:

The 96-h LC50 value in fish (Cyprinus carpio) is 0.43 mg/L.

Executive summary:

A study was performed to assess the acute toxicity of the substance to fish (Cyprinus carpio). The study was conducted in accordance with OECD Guideline for Testing of Chemicals No. 203. In a semi-static test set-up in airtight closed 3.2-L vessels seven fish per concentration were exposed for 96 hours to a Water Soluble Fraction (WSF) prepared at a loading rate of 100 mg/L and 5 dilutions of this WSF. Throughout the test, test solutions were refreshed in intervals of 24 hours. Clinical effects (such as hypoactive swimming, loss of equilibrium or immobility) and incidences of mortality were recorded for the control and each test group at t = 3, 24, 48 and 72 and 96 hours. Throughout the 24h renewal intervals, the measured concentration decreased at max. to 2.8 – 48% of initial. Therefore average exposure concentrations were calculated and the LC50 was calculated based on these concentrations.

Description of key information

A study was performed to assess the acute toxicity of the substance to fish (Cyprinus carpio). The study was conducted in accordance with OECD Guideline for Testing of Chemicals No. 203. In a semi-static test set-up in airtight closed 3.2-L vessels seven fish per concentration were exposed for 96 hours to a Water Soluble Fraction (WSF) prepared at a loading rate of 100 mg/L and 5 dilutions of this WSF. Throughout the test, test solutions were refreshed in intervals of 24 hours. Clinical effects (such as hypoactive swimming, loss of equilibrium or immobility) and incidences of mortality were recorded for the control and each test group at t = 3, 24, 48 and 72 and 96 hours. Throughout the 24h renewal intervals, the measured concentration decreased at max. to 2.8 – 48% of initial. Therefore average exposure concentrations were calculated and the LC50 was calculated based on these concentrations.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.43 mg/L