Methyl 3-oxo-2-pentylcyclopentaneacetate

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 04 August 1998 to 23 October 1998

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Non GLP test comparable to OECD Guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Methyl 3-oxo-2-pentylcyclopentaneacetate

EC Number:

246-495-9

EC Name:

Methyl 3-oxo-2-pentylcyclopentaneacetate

Cas Number:

24851-98-7

Molecular formula:

C13H22O3

IUPAC Name:

methyl 3-oxo-2-pentylcyclopentaneacetate

Details on test material:

- Name of test material (as cited in study report): Methyl dihydrojasmonate
- Substance type: pure active substance
- Physical state: liquid
- Analytical purity: >99.98 % (sum of 2 isomers and of few position isomers)
- Stability under test conditions: stable
- Storage condition of test material: room temperature in the dark
- Other: test material is moderately soluble (water solubility > 500 mg/l @ 20°C), has a low Vapour pressure (< 0.0001 Pa) and a logPow < 3

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not Applicable

Sampling and analysis

Analytical monitoring:

no

Details on sampling:

Not applicable

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 2 g of the carrier solvent (HCO-40) was added to 1000 mg of the test substance and mixed well. Then the mixture was filled up to 1000 mL with purified water in order to prepare a stock solution containing the test substance at the concentration of 1000 mg/L. At the same time, another stock solution containing only 2 g of carrier solvent (HCO-40) was prepared in the same manner. The test solutions were prepared by filling up a specific volume of the stock solutions to 5.0 L with dilution water (10 ml for 2.0 mg/ml, 21 ml for 4.2 mg/ml, 44.5 ml for 8.9 mg/ml, 95 ml for 19 mg/ml and 200 ml for 40 mg/ml).
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): HCO-40 (emulsifier)
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): Stock solution: 2g/l; Final test solutions: from 0.1 mg/l (for 2mg/l test concentration) to 80 mg/l (for 40 mg/ml test concentration and solvent control)
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none

Test organisms

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Orange killifish
- Source: Sankyosuisan (1-1, Ichigayatamachi, Shinjuku-ku, Tokyo - Japan
- Age at study initiation (mean and range, SD): young
- Length at study initiation (length definition, mean, range and SD): 2.23 cm (1.93-2.56 cm), n=20
- Weight at study initiation (mean and range, SD): 0.189 g (0.114-0.289 g), n=20
- Feeding during test: No. Feeding was stopped 24 hours before the start of the exposure



ACCLIMATION
- Acclimation period: > 7 days
- Acclimation conditions (same as test or not): yes
- Type and amount of food: Commercially available Tetramin
- Health during acclimation (any mortality observed): mortality rate during 7 days before the start of the exposure was less than 5%

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not applicable

Test conditions

Hardness:

49 mg CaC03/l

Test temperature:

24 ± 1 °C (See table 3 in Any other information on results incl. tables)

pH:

7.1 to 7.7 (See table 3 in Any other information on results incl. tables)

Dissolved oxygen:

8.2 to 5.5 mg/l (i.e. not less than 60% of the air saturation as O2 saturation at 24°C correspond to 8.25mg/l)
(See table 3 in Any other information on results incl. tables)

Salinity:

Not applicable

Nominal and measured concentrations:

Preliminary range-finding: 0 (control), 0 (solvent control), 3.0, 10 and 30 mg/l
Test: 0 (control) , 0 (solvent control), 2.0, 4.2, 8.9, 19, 40 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): 5.0 liter glass beakers
- Material, size, headspace, fill volume: 5 liter glass beaker (open system)
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): semi-static system with batchwise renewal of the test solution every 24 hours.
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: 0.378 g/l


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated drinking water
- Conductivity: not required for drinking water

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod: 16h light/ 8h dark


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortalities were recorded after 24, 48, 72 and 96 hours. Symptoms of toxicity observed and abnormal responses of the fish were also recorded. Generic terms and definitions for toxic cases are shown as follows:
Dead Fish are considered dead if touching produces no reaction.
Abnormal respiration: Observable difference in gill movements by comparison with the control.
Abnormal swimming: Observable clear difference in swimming behavior by comparison with the control. e.g. slow, irritated, spasm, inverted etc.
Loss of swimming capacity: Moving at the bottom of the surface, but being impossible to swim. Including “on side swimming ” and “suspended animation”
Remark: Dead fishes were removed as soon as possible in order to keep the test water in good quality, when they were observed. Mortalities were recorded.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.1
- Justification for using less concentrations than requested by guideline: not applicable
- Range finding study
- Test concentrations: 3.0, 10 & 30 mg/l
- Results used to determine the conditions for the definitive study: mortalities at 96 hours were 0 % for 3.0 mg/L treatment; 20 % for the 10 mg/L
treatment; and 100 % for the 30 mg/L treatment. In accordance with the above mentioned preliminary test result, 5 (five) series of test concentration: 2.0, 4.2, 8.9, 19 and 40 mg/L (common ratio: 2.1) were determined for the main test.

Reference substance (positive control):

yes

Remarks:

Copper(II) sulfate pentahydrate reagent-grade: LC50: 0.73 mg/l (95% CL: 0.53-1.0 mg/l)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

28 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 19-40 mg/l

Effect concentrations 2

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

23 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 8.9-40 mg/l

Effect concentrations 3

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

21 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 8.9-40 mg/l

Effect concentrations 4

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

19 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 8.9-40 mg/l

Effect concentrations 5

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: behavior/mobility

Details on results:

- Behavioural abnormalities: Abnormal swimming and the loss of swimming capacity were observed at 4.2 mg/L and higher. Paralysis was observed at 8.9 mg/l and higher. No abnormal responses were observed in the Control (blank) and the Solvent Control.
- Mortality of control: no mortality observed both in the control and in the solvent control
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none

Results with reference substance (positive control):

- Results with reference substance valid? YES (Copper(II) sulfate pentahydrate reagent-grade)
- LC50: 0.73 mg/l (95% CL: 0.53-1.0 mg/l)

Reported statistics and error estimates:

The cumulative percentage mortality for each treatment was calculated, and the Median Lethal Concentration (LC50) was calculated by the Binomial theory. The 95% confidence limits for the calculated LC50 values are also determined.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Table 1: Cumulative Mortality data

Nominal Concentration (mg/l)	Cumulative mortality (initial population : 10)
	24h	48h	72h	96h
Control	0	0	0	0
Solvent Control	0	0	0	0
2.0	0	0	0	0
4.2	0	0	0	0
8.9	0	0	0	0
19	0	3	4	5
40	10	10	10	10

 

Table 2: Behavioral Observations

Concentration (mg/l)	Time
	24h	48h	72h	96h
Control	N	N	N	N
Solvent Control	N	N	N	N
2.0	N	N	N	N
4.2	N	AS-1	AS-3	AS-3
8.9	AS-10	AS-10	AS-8 AQ-2	AS-8 AQ-2
19	AS-7 AQ-3	AS-4 AQ-3	AS-2 AQ-4	AS-1 AQ-4
40	A/D	A/D	A/D	A/D
N:        No toxicological effects AS:      Abnormal swimming AQ:     Paralysis A/D:    All fish dead

 

Table 3: Physico-chemical measurements

	Temperature (°C)
Nominal Concentration (mg/l)	0 hours	24 hours		48 hours		72 hours		96 hours
	new	old	new	old	new	old	new	old
Control	23.5	24.2	24.3	24.3	24.1	24.0	24.5	24.0
Solvent control	23.5	24.2	24.3	24.3	24.1	24.0	24.5	24.0
2.0	23.5	24.1	24.3	24.3	24.1	24.0	24.5	24.0
4.2	23.5	24.1	24.3	24.3	24.1	24.0	24.5	24.0
8.9	23.5	24.1	24.3	24.3	24.1	24.0	24.5	24.0
19	23.5	24.1	24.2	24.2	24.1	24.0	24.5	24.0
40	23.5	--	--	--	--	--	--	--
	Dissolved Oxygen Concentration (mg/l)
	new	old	new	old	new	old	new	old
Control	8.2	6.8	8.1	6.1	8.1	6.9	7.9	6.3
Solvent control	8.1	6.3	8.0	6.1	8.1	6.2	7.9	6.1
2.0	8.1	7.2	8.2	6.5	8.1	6.7	8.0	6.5
4.2	7.8	7.1	8.2	6.2	8.1	6.7	8.1	6.2
8.9	7.9	7.0	8.2	6.1	8.2	6.6	7.9	5.8
19	7.9	6.4	8.1	5.5	8.2	5.8	7.6	5.5
40	7.9	--	--	--	--	--	--	--
	pH
	new	old	new	old	new	old	new	old
Control	7.7	7.5	7.6	7.4	7.5	7.3	7.5	7.1
Solvent control	7.7	7.5	7.7	7.3	7.5	7.3	7.6	7.2
2.0	7.7	7.5	7.7	7.3	7.6	7.3	7.6	7.2
4.2	7.7	7.5	7.7	7.3	7.6	7.3	7.6	7.2
8.9	7.7	7.4	7.7	7.2	7.5	7.2	7.6	7.2
19	7.7	7.4	7.7	7.2	7.6	7.2	7.6	7.1
40	7.7	--	--	--	--	--	--	--
Legend: New: freshly prepared test solutions Old: test solutions after 24 hours exposure -- No measurement as all fishes were dead at this observation time

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

The test compound concentration in solution was nominal and not analytically determined. However, as the system was semi-static (daily complete renewal of the test solution), it can be assumed that the test compound concentration was maintained (≥80%).

Conclusions:

The acute toxicity of the test material to the Orange Killifish (Oryzias latipes) has been investigated and gave a 96-Hour LC50 of 19 mg/L and a NOEC of 2.0 mg/L.

Executive summary:

Introduction. Acute toxicity test of Methyldihydrojasmonate to Orange killifish (Oryzias latipes) was determined according to the OECD Guideline for testing of chemicals No. 203 "Fish, Acute Toxicity Test" (1992).

 

Methods.The test was performed in Semi-static conditions with a daily batchwise renewal of the test solution as the test material is expected to be stable in the test solution as shown in the study hydrolysis as a function of pH performed on the same test material (OECD TG 111, half life pH 7 > 1 year). The substance has been shown to be moderately soluble (>500 mg/l), and not volatile in the physico-chemical endpoints/studies. Therefore nominal concentrations were used to determine NOEC and EC50 values.

Preliminary range-finding test was made with three concentration series of 3.0, 10 and 30 mg/l (5 fishes for each treatment) before the main test. As a result, mortalities at 96 hours are 0 % for the 3.0 mg/l treatment; 20 % for the 10 mg/l treatment; and 100 % for the 30 mg/l treatment. In accordance with the above mentioned preliminary test result, 5 (five) series of test concentration, 2.0, 4.2, 8.9, 19 and 40 mg/L (common ratio: 2.1) were determined for the main test. A control and a solvent control were also performed as test compound was solubilized in HCO-40.

 

Results.

The minimum concentration causing 100 % mortality within the period of the test (96 hours) was 40 mg/L. No mortality was observed in the Control (blank) and the Solvent Control during the test period. Abnormal swimming and the loss of swimming capacity were observed at 4.2 mg/L and higher tested concentration. No abnormal responses were observed in the Control (blank), the Solvent Control and at 2.0 mg/l. Therefore, the No Observed Effect Concentration (NOEC) was 2.0 mg/l.

The median lethal concentration (LC50) of every 24-hour, determined by the geometric mean of the highest concentration causing no mortality and the lowest concentration producing 100% mortality, is shown as follow:

 

Exposure period	LC50 (mg/l)	95% Confidence Limits (mg/l)
24 48 72 96	28 23 21 19	19 -40 8.9 -40 8.9 -40 8.9 -40

 

The 96-Hour LC50 based on nominal test concentrations was equals 19 mg/L and the corresponding No Observed Effect Concentration (NOEC) was 2.0 mg/L.