(2E)-2-ethyl-4-(2,2,3-trimethylcyclopent-3-en-1-yl)but-2-en-1-ol

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:

1999

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions. The measured concentrations were outside the 80-120% range. Therefore the results were expressed as means of measured concentrations.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Deviations:

yes

Remarks:

The measured concentrations were outside the 80-120% range

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0, 0.38, 0.75; 1.5, 3.0, 6.0 mg/L
- Sampling method: Each control and test substance treatment replicate was sampled and the samples were composited by treatment level. A single sample was collected from the diluter mixing box. Samples were collected subsurface from the approximate center (about mi-depth) of test chamber and the mixing box.
Quality control fortifications samples (spikes) were prepared by spiking 10.0 mL of control water with Super Radjanol. Spikes were prepared at concentrations braketing the range of nominal concentrations of the test substance treatments.
Samples at concentration below 0.38 mg/L were diluted with acetonitrile to a final concentration of 50%; samples above 0.75 mg/L were diluted in 80:20 acetonitrile: ABC reagent water (v:v) such that the concentration of analyte in the final extracts was within the range of the standard reference curve.

Vehicle:

yes

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: Bluegill sunfish
- Source: Osage Catfisheries
- Length at study initiation (length definition, mean, range and SD): 30 ± 4.1 mm
- Weight at study initiation (mean and range, SD): 0.618 ± 0.347 g

ACCLIMATION
- Acclimation period: 108 days prior to definitive test initiation
- Acclimation conditions (same as test or not):
- Type and amount of food: brine shrimp nauplii, salmon starter, fish flake food
- Feeding frequency: daily during the holding period
- Health during acclimation (any mortality observed): no mortality observed

QUARANTINE (wild caught)
- Duration: 48 hours prior to test initiation
- Health/mortality: no mortality observed

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

no post exposure observation period.

Hardness:

144 mg/L as CaCO3 at day 0

Test temperature:

22 ± 1 °C

pH:

8.1 to 8.3 in all control and test substance treatment solutions during the test

Dissolved oxygen:

from 7.9 to 8.0 mg/L at test initiation
≥ 7.4 mg/L for the duration of the test

Nominal and measured concentrations:

The results were based on measured concentrations

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquaria
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: volume of approximatively 15 L (34 cm (lenght) x 22cm (width) x 30cm (height))
- Type of flow-through (e.g. peristaltic or proportional diluter): proportional diluter
- Renewal rate of test solution (frequency/flow rate): 3.8 tank volume replacements per chamber per day
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 2
- Biomass loading rate: 0,412 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: moderately hard laboratory freshwater prepared by blending naturally hard well water with well that was demineralized by reverse osmosis
- Total organic carbon: 0 mg/L
- Metals: < 0.5 mg/L
- Pesticides: < 1µg/L
- Alkalinity: 150 mg/L as CaCO3
- Conductivity: 323 µS/cm


OTHER TEST CONDITIONS
- Adjustment of pH: no adjustement
- Photoperiod: 16-hour light and 8-hour dark photoperiod with 30 minutes simulated dawn and dusks period
- Light intensity: 820.5 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and sublethal parameters

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study: conducted under static conditions at nominal concentrations of 0.0 (control), 0.0 (vehicle control), 1.0, 10.0, 20.0, 40.0 mg a.i/L. Three fish were tested per treatment. After 96 hours of exposure, mortality was 100% at test concentrations > 10 mg a.i/L. Mortality was O and 33% in the dilution water and vehicle controls, respectively.
- Test concentrations: 0, 0.38, 0.75, 1.5, 3, 6

See Table 1: Measured concentrations of Super Radjanol in samples taken during a 96-hour flow-through acute test with the Bluegill Sunfish (Lepomis macrochirus)

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.1 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 0.89-1.2 mg/L

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.49 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

See following Tables in "Any other information on results" section:
Table 1: Mortality of Bluegill Sunfish (Lepomis macrochirus) during a 96-hour exposure to Super Radjanol under flow-through test conditions
Table 2: Calculated LC50 values, based upon mean measured concentrations, for a 96-hour flow-through exposure of Bleugill sunfish (Lepomis macrochirus) to Super Radjanol

Results with reference substance (positive control):

no data on positive control

Reported statistics and error estimates:

no information reported

Sublethal observations / clinical signs:

Table 1: Mortality of Bluegill Sunfish (Lepomis macrochirus) during a 96-hour exposure to the test item under flow-through test conditions

Cumulative number dead
Mean measured conc. (mg a.i/L)	Rep	6-h	24-h	48-h	72-h	96-h	Treatment mortality (%)
Control	A	0	0	0	0	0	0
	B	0	0	0	0	0
Vehicle control	A	0	0	0	0	0	0
	B	0	0	0	0	0
0.38	A	0	0	0	0	0	0
	B	0	0	0	0	0
0.49	A	0	0	0	0	0	0
	B	0	0	0	0	0
0.99	A	0	0	0	1	3	45
	B	0	0	0	1	6
2.2	A	0	1	9	10	10	100
	B	0	2	10	10	10
6.0	A	4	10	10	10	10	100
	B	3	10	10	10	10

 

 

Table 2: Calculated LC₅₀ values, based upon mean measured concentrations, for a 96-hour flow-through exposure of Bluegill sunfish (Lepomis macrochirus) to the test item

 

Exposure period (hour)	LC50(mg a.i/L; ppm)	95-percent confidence limits	Slope	Statistical method
24	3.2	2.7-3.7	NC*	Spearman-karber
48	1.5	1.4-1.7	NC*	Spearman-karber
72	1.4	1.2-1.5	NC*	Spearman-karber
96	1.1	0.89-1.2	NC*	Spearman-karber

* NC: The slope could not be calculated (NC) using the Spearman-Karber method

Validity criteria fulfilled:

yes

Conclusions:

The 96-hour LC50 value is 1.1 mg a.i/L with 95% confidence limits of 0.89 to 1.2 mg a.i/L (measured concentrations)
The 96-hour NOEC value is 0.49 mg a.i/L (measured concentrations)

Executive summary:

In a 96-h acute toxicity study, Bluegill sunfish (Lepomis macrochirus) were exposed to the test item at measured concentrations of 0 (control, vehicle control), 0.38, 0.49, 0.99, 2.2, 6 mga.i./L under flowthrough conditions. The 96-h LC₅₀ was 1.1 mg a.i./L. The NOEC value, based on mortality/sublethal effects, was 0.49 mg a.i./L. Sublethal effects of loss of equilibrium and irregular respiration were observed in the groups exposed to concentrations > 0.99 mg a.i/L of the test item.

This toxicity study is classified as acceptable and satisfies with the guidelines requirements for OECD 203, fish acute toxicity study.

 

Results Synopsis

 

Test organism size/age (mean wet weight or length): Bluegill Sunfish (Lepomis macrochirus); mean weight = 0.618 ± 0.347 g; mean length = 30 ± 4.1 mm

Test Type (Flowthrough, Static, Static Renewal): flow-through

 

LC₅₀: 1.1 mg a.i./L             95% C.I.: 0.89 to 1.2 mg a.i./L}

NOEC: 0.49 mg a.i./L} 

Endpoint Effected: mortality

Description of key information

The substance exhibits a LC50 for freshwater fish of 1.1 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1.1 mg/L

Additional information

Two experimental data and one QSAR data are available to determine the short-term toxicity to aquatic invertebrates.

Kranzfelder, J.A, 1999

In an acute aquatic toxicity study performed according to the EPA OPPTS 850.1075 guideline, groups of Bluegill sunfish (Lepomis macrochirus) were exposed to Super Radjanol at concentrations of 0, 0.38, 0.75, 1.5, 3.0 or 6.0 mg/L (nominal) for 96 hours under flow-through conditions. Mortality, behaviour and morphological aberrations were observed at 6, 24, 48, 72 and 96 hours. Mortalities (96-hour exposure) at 0, 0.38, 0.49, 0.99, 2.2 or 6.0 mg/L (mean of concentrations measured at 0, 48 and 96 h) were 0, 0, 0, 45, 100 and 100%, respectively. Calculated LC50 at 24, 48, 72 and 96 hours were 3.2, 1.5, 1.4, 1.1 mg/L respectively, based on measured concentration of test item. This toxicity study is classified as acceptable and satisfies the guideline requirements for an acute fish toxicity study. This study is given a reliability score of 2 (reliable with restrictions) since the test method is considered as an alternative to the EU test method and the OECD test guideline.

Thomas, 2014

A QSAR prediction was performed to assess the acute toxicity of the test item 2-ethyl-4-(2,2,3-trimethyl-3-cyclopenten-1-yl)-(2E)-buten-1-ol to fish. This QSAR has been validated to be compliant with the OECD recommendations for QSAR modeling (OECD, 2004) and predicts the endpoint value which would be expected when testing the substance under experimental conditions in a laboratory following Guideline for Testing of Chemicals No. 203, "Fish Acute Toxicity Test" (1), referenced as Method C.1 of Commission Regulation No. 440/2008 (2). The criterion predicted was the LC50(Median Lethal Concentration), a statistically derived concentration which is expected to cause mortality in 50% of test animals within a period of 96 hours. The QSAR is based on validated data for a training set of 67 chemicals derived from 96-hour test on fish, for which the concentrations of the test item had been determined by chemical analyses over the test period.