(1R,5R)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene

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:

1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Study conducted similarly to OECD guideline 203 but without GLP compliance and with minor deviations (no data on acclimation period). The report is sufficiently well documented.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

No data on acclimation period

GLP compliance:

not specified

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0, 130, 260, 390, 520 and 650 μg/L
- Sampling time: 0, 24, 48, 72 and 96 hours

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Test solutions was provided as liquid/liquid saturation units by adding excess of test chemical in 14 L of Lake Superior water
- Controls: Dilution water
- Chemical name of vehicle: water
- Concentration of vehicle in test medium: Stock (650 µg/L liq-liq equil.)

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead minnows
- Source: ERL-Duluth, USA
- Age at study initiation: 30 days
- Length at study initiation: 20.9 mm (SD=2.2)
- Weight at study initiation: 128 mg (SD=35)

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None

Hardness:

45.2 mg CaCO3/L (SD=0.3)

Test temperature:

25.2ºC (SD=0.2)

pH:

7.7 (SD=0.2)

Dissolved oxygen:

6.7 mg O2/L (SD=0.7)

Salinity:

42.2 mg CaCO3/L (SD=0.9)

Nominal and measured concentrations:

Nominal concentrations: 0, 130, 260, 390, 520 and 650 μg/L
Average measured concentrations: <31, 58, 140, 229, 303 and 424 μg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Six crystallizing dishes were used as test chambers in the electronic diluter with tank volumes of 200 mL; test solution depth: 4-5 cm
- Type (delete if not applicable): Closed
- Type of flow-through: Electronic diluter
- Renewal rate of test solution (frequency/flow rate): Flow rates of 21 mL/min to each tank produced toxicant concentrations of 58-424 μg/L
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): One
- No. of vessels per vehicle control (replicates): One
- Biomass loading rate: 6.4 g/L

TEST MEDIUM / WATER PARAMETERS
- Source of dilution water: Lake Superior water
- Preparation of dilution water: Water was passed through sand filters, ultraviolet sterilizers and 55 µm fibre filter. Test waters were then aerated and heated at 25 ± 1 °C
- Intervals of water quality measurement: Tank temperatures (ºC), dissolved oxygen (mg/L) and pH were measured daily in each of the test chambers where test organisms were still alive

OTHER TEST CONDITIONS
- Photoperiod: 16 hour/day light cycle; cool white fluorescent light
- Light intensity: 12-28 lumens

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Behavior and morphological aberrations were observed at 0, 24, 48, 72 and 96 hours

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

280 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 259-303 μg/L

Key result

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

179 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: Affected fish swam near the tank bottom, were hypoactive and underreactive to external stimuli, exhibited hemorrhaging and lost equilibrium prior to death.
- Mortality (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L (measured) were 0, 0, 0, 0, 80 and 100%, respectively.
- Effect (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L μg/L (measured) were observed in 0/10, 0/10, 0/10, 10/10, 10/10 and 10/10 fish respectively.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Measured concentrations were lower than nominal concentrations due to volatility.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

- LC50 or EC50 values were calculated using the corrected average of the analyzed tank concentrations and the Trimmed Spearman-Karber Method (Hamilton et al., 1977).
- Mean toxicant concentrations used in the calculations were corrected for analytical spike recoveries

Sublethal observations / clinical signs:

Table 1: Mortalities

 

Hour	Control (0 µg/L)	A (130 µg/L)	B (260 µg/L)	C (390 µg/L)	D (520 µg/L)	E (650 µg/L)
Initial	10	10	9	10	10	10
24	0	0	0	0	7	8
48	0	0	0	0	8	10
72	0	0	0	0	8	10
96	0	0	0	0	8	10

Table 2: Effects

Hour	Control (0 µg/L)	A (130 µg/L)	B (260 µg/L)	C (390 µg/L)	D (520 µg/L)	E (650 µg/L)
Initial	10	10	9	10	10	10
24	0	0	0	10	10	10
48	0	0	0	10	10	10
72	0	0	0	10	10	10
96	0	0	0	10	10	10

 

Validity criteria fulfilled:

yes

Remarks:

(mortality in control < 10%; flow-through test conditions; dissolved O2 concentration > 60% of air saturation value; results based on measured concentrations)

Conclusions:

The 96h-LC50 of d-alpha pinene to fathead minnow (Pimephales promelas) was calculated to be 280 (259 -303) μg/L.

Executive summary:

In an acute aquatic toxicity study performed similarly to OECD guideline 203, groups (10/dose) of fathead minnows (Pimephales promelas) were exposed to d-alpha pinene at concentrations of <31, 58, 140, 229, 303 and 424 μg/L (measured) for 96 hours under flow-through conditions. Mortality, behaviour and morphological aberrations were observed at 24, 48, 72 and 96 hours. All validity criteria were fulfilled.Mortalities (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L (measured) were 0, 0, 0, 0, 80 and 100%, respectively. Treatment-related effects (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L (measured) were observed in 0/10, 0/10, 0/10, 10/10, 10/10 and 10/10 fish respectively. The 96 hour LC50, EC50 and NOEC of d-alpha pinene to fathead minnow (Pimephales promelas) were calculated to be 280 (259 -303) μg/L, 179 μg/L and 140 μg/L, respectively.

 

 

 

Description of key information

Key study: Test method similar to OECD TG 203. The 96h-LC50 of d-alpha pinene to fathead minnow (Pimephales promelas) was calculated to be 280 (259 -303) μg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.28 mg/L

Additional information

Key study: In an acute aquatic toxicity study performed similarly to OECD guideline 203, groups (10/dose) of fathead minnows (Pimephales promelas) were exposed to d-alpha pinene at concentrations of <31, 58, 140, 229, 303 and 424 μg/L (measured) for 96 hours under flow-through conditions. Mortality, behaviour and morphological aberrations were observed at 24, 48, 72 and 96 hours. All validity criteria were fulfilled. Mortalities (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L (measured) were 0, 0, 0, 0, 80 and 100%, respectively. Treatment-related effects (96 hour) at <31, 58, 140, 229, 303 and 424 μg/L (measured) were observed in 0/10, 0/10, 0/10, 10/10, 10/10 and 10/10 fish respectively. The 96 hour LC50, EC50 and NOEC of d-alpha pinene to fathead minnow (Pimephales promelas) were calculated to be 280 (259 -303) μg/L, 179 μg/L and 140 μg/L, respectively.