Cyclopropanecarboxylic acid, 2-[1-(3,3-dimethylcyclohexyl)ethoxy]-2-methylpropyl ester

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:

24-05-2004 to 02-08-2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study performed under GLP. All relevant validity criteria were met with acceptable deviations.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

No concurrent positive control conducted as part of the study. A non-concurrent reference test utilising pentachlorophenol (PCP) included in the study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

yes

Remarks:

No concurrent positive control conducted as part of the study. A non-concurrent reference test utilising pentachlorophenol (PCP) included in the study

GLP compliance:

yes (incl. QA statement)

Remarks:

inspected: May 2002 ; signature: September 2002

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0 (control), 0.5, 0.8, 1.3, 2.0, 3.2 mg/L nominal concentration
Analysis of the samples taken at the start of the final test showed measured concentrations of 1.2, 1.7 and 1.8 mg/L at the nominal concentrations of 1.3, 2.0 and 3.2 mg/L, respectively. During the remainder of the test analysis was only performed on samples taken from nominally 3.2 mg/L. Results of the 24-hour old sample taken after 24 hours of exposure showed a measured concentration of 1.1 mg/L. Measured concentrations during the last renewal interval (t=72-96 hours) showed a measured concentration that decreased from 2.7 to 2.0 mg/L. The geometric average exposure concentration at nominal 3.2 mg/L was calculated to correspond to 1.9 mg/L.
- Sampling method: From all freshly prepared solutions were sampled at t=0 and t=72 hours and from all the 24h-old solutions at t=24 hours. Finally, from the 24h-old solutions of the control and 3.2 mg/L at the end of the test period. Control, 0.5, 0.8 and 1.3 mg/L: 4.5 mL 2.0 and 3.2 mg/L: 1.5 mL. All samples were taken from the approximate centre of the test vessels. Samples taken at t=0, 24 and 72 hours of exposure were stored in a refrigerator until analysis.
- Sample storage conditions before analysis: See above.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Preparation of test solutions started with stock solutions in acetone at a factor of 10,000 higher than the final test solutions. Volumes of 100 µL were then added per litre ISO-medium to obtain the desired concentration. The highest concentration prepared during the range-finding test was then magnetically stirred for 40 minutes, while the two lower concentrations were stirred for 10 minutes after which the respective solutions were used for testing. Preparation during the final test included a magnetic stirring period of 30 minutes, which was followed by a stabilisation period of 20 to 30 minutes, except for the two lowest concentrations that were fully dissolved. The water phase was collected in case a test substance film was observed after the stabilisation period. All final test solutions were all clear and colourless except the highest concentration tested in the range-finding test (5.0 mg/L nominal), which contained a thin floating layer. Test solutions were daily renewed during the final test.
Analysis of the samples taken at the start of the final test showed measured concentrations of 1.2, 1.7 and 1.8 mg/L at the nominal concentrations of 1.3, 2.0 and 3.2 mg/L, respectively. During the remainder of the test analysis was only performed on samples taken from nominally 3.2 mg/L. Results of the 24-hour old sample taken after 24 hours of exposure showed a measured concentration of 1.1 mg/L. Measured concentrations during the last renewal interval (t=72-96 hours) showed a measured concentration that decreased from 2.7 to 2.0 mg/L. The geometric average exposure concentration at nominal 3.2 mg/L was calculated to correspond to 1.9 mg/L.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Acetone
- Concentration of vehicle in test medium (stock solution and final test solution): < 0.01% v/v
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No precipitate reported within the final test at up to the highest test item concentration.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Common Carp (Cyprinus carpio)
- Strain: Not reported.
- Source: Recognised supplier (listed in full study report).
- Age at study initiation (mean and range, SD): Not reported.
- Length at study initiation (length definition, mean, range and SD): 2.3 ± 0.2 cm (definitive test)
- Weight at study initiation (mean and range, SD): 0.34 ± 0.10 g (definitive test) i.e. ca. 0.53 g bw/L loading rate
- Method of breeding: Not reported. F1 from a single parent-pair bred in UV-treated water.
- Maintenance of the brood fish: Not reported. See above.

ACCLIMATION
- Acclimation period: > 12 days.
- Acclimation conditions (same as test or not): Yes. ISO-medium utilised.
- Type and amount of food during acclimation: certified pelleted diet (documented in the full study report).
- Feeding frequency during acclimation: daily (until 48 hours prior to test).
- Health during acclimation (any mortality observed): < 5% mortality in 7 days pre-test

QUARANTINE (wild caught)
- Duration: Not applicable.
- Health/mortality: Not applicable.

FEEDING DURING TEST
- Food type: Not applicable.
- Amount: Not applicable.
- Frequency: Not applicable.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

According to OECD TG 203 guidelines.

Post exposure observation period:

none.

Hardness:

ca. 250 mg/L (expressed as CaCO3)

Test temperature:

14 ºC to 14.5 ºC

pH:

7.8-7.9 fresh media); 7.3-7.7 (expired media)

Dissolved oxygen:

9.0-9.3 mgO2/L (fresh media); 7.3-8.1 mgO2/L (expired media)

Salinity:

not applicable.

Nominal and measured concentrations:

Concentrations: 0 (control), 0.5, 0.8, 1.3, 2.0, 3.2 mg/L nominal concentration
Analysis of the samples taken at the start of the final test showed measured concentrations of 1.2, 1.7 and 1.8 mg/L at the nominal concentrations of 1.3, 2.0 and 3.2 mg/L, respectively. During the remainder of the test analysis was only performed on samples taken from nominally 3.2 mg/L. Results of the 24-hour old sample taken after 24 hours of exposure showed a measured concentration of 1.1 mg/L. Measured concentrations during the last renewal interval (t=72-96 hours) showed a measured concentration that decreased from 2.7 to 2.0 mg/L. The geometric average exposure concentration at nominal 3.2 mg/L was calculated to correspond to 1.9 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: 6.5 L for test and control vessels, glass
- Type (delete if not applicable): closed, covered to reduce evaporation
- Material, size, headspace, fill volume: 4.5 L fill volume test media per test vessel
- Aeration: No.
- Renewal rate of test solution (frequency): 24 h
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): Not applicable
- Biomass loading rate: 0.53 g/L (mean)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Drinking water; chlorine removed by activated carbon filtration and partial softening (DI).
- Total organic carbon: Not reported
- Particulate matter: Not reported
- Metals: Not reported
- Pesticides: Not reported
- Chlorine: Not reported
- Alkalinity: Not reported
- Ca/mg ratio: Not reported
- Conductivity: Not reported
- Culture medium different from test medium: No.
- Intervals of water quality measurement: 24 hours.

OTHER TEST CONDITIONS
- Adjustment of pH: No.
- Photoperiod: 16 h light / 8 h darkness
- Light intensity: Not reported.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and visible abnormality recorded at 2.5 h, 24 h, 48 h, 72 h and 96 h.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Concentrations justified from the results of the range finding study.
- Range finding study: yes (semi-static) ; 3 organisms per concentration.
- Test concentrations: blank control, 0.05, 0.5 and 5.0 mg/L nominal
- Results used to determine the conditions for the definitive study: Yes. No mortalities at 0 (control), 0.05 or 0.5 mg/L. 67% mortality at 5.0 mg/L nominal ; an oil like floating layer was observed at the highest nominal concentration throughout the test period.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1.9 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: (95% CL: - mg/L) ; geometric mean measured average concentrations ; LC50 was exceeding limit of solubility

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1.9 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

other: Mortality and behaviour

Remarks on result:

other: NOEC and LC50 was exceeding limit of solubility

Details on results:

- Behavioural abnormalities: No mortality or clinical effects were observed in any of the concentrations tested.
- Observations on body length and weight: Not reported
- Other biological observations: Not reported
- Mortality of control: 0%
- Other adverse effects control: No abnormalities observed in the control.
- Abnormal responses: None reported.
- Effect concentrations exceeding solubility of substance in test medium: Yes. Test solutions were daily renewed during the final test. Analysis of the samples taken at the start of the final test showed measured concentrations of 1.2, 1.7 and 1.8 mg/L at the nominal concentrations of 1.3, 2.0 and 3.2 mg/L, respectively. During the remainder of the test analysis was only performed on samples taken from nominally 3.2 mg/L. Results of the 24-hour old sample taken after 24 hours of exposure showed a measured concentration of 1.1 mg/L. Measured concentrations during the last renewal interval (t=72-96 hours) showed a measured concentration that decreased from 2.7 to 2.0 mg/L. The geometric average exposure concentration at nominal 3.2 mg/L was calculated to correspond to 1.9 mg/L.

Results with reference substance (positive control):

- Results with reference substance valid? : Yes.
- Mortality: at 0.1, 0.22 and 0.46 mg/L PCP nominal concentrations: 0, 0 and 100% mortality respectively at 96 h
- LC50: 96h-LC50 = 0.32 (C.I. 0.22 – 0.46) mg/L
- Other: range of the 96hLC50 for species is generally between 0.10 and 0.46 mg/L based on HCD documented in the full study report.

Reported statistics and error estimates:

Not applicable.

Sublethal observations / clinical signs:

Table 1.0 - Cumulative mortality

Nominal Concentration (mg/L)	Cumulative mortality (n=10)						Total mortality %
	2.5h	6h	24h	48h	72h	96h
0.0 (control)	0	-	0	0	0	0	0
0.5	0	-	0	0	0	0	0
0.8	0	-	0	0	0	0	0
1.3	0	-	0	0	0	0	0
2.0	0	-	0	0	0	0	0
3.2 #1	0	-	0	0	0	0	0

#1 : geometric mean measured average concentration : 1.9 mg/L
- : not recorded

Validity criteria fulfilled:

yes

Conclusions:

The 96 hour LC50 for the test item to Common Carp (Cyprinus carpio) was determined to be > 1.9 (C.I. - ) mg/L based on geometric mean measured average concentrations.No effects seen at solubility limit in the test medium.

Executive summary:

The acute toxicity of the test item to Common Carp (Cyprinus carpio) was determined in a 96 hour semi-static test according to OECD TG 203 and EU Method C.1 guidelines under GLP. Following a preliminary range-finding test, fish were exposed, in groups to an aqueous solution of the test item at 0.5, 0.8, 1.3, 2.0 and 3.2 mg/L nominal concentrations with daily renewal. Samples for analytical confirmation of actual exposure concentrations were taken from freshly prepared solutions at the start and after 72 hours of exposure and from 24-hour old solutions after 24 and 96 hours of exposure. Analysis of the samples taken at the start of the final test showed measured concentrations of 1.2, 1.7 and 1.8 mg/L at the nominal concentrations of 1.3, 2.0 and 3.2 mg/L, respectively. During the remainder of the test analysis was only performed on samples taken from nominally 3.2 mg/L. Results of the 24-our old sample taken after 24 hours of exposure showed a measured concentration of 1.1 mg/L. Measured concentrations during the last renewal interval (t=72-96 hours) showed a measured concentration that decreased from 2.7 to 2.0 mg/L. The average exposure concentration at nominal 3.2 mg/L was calculated to correspond to 1.9 mg/L. Analytical results were in agreement with the results obtained during the range-finding test. This confirmed the maximum solubility in ISO-medium was at approximately 2.0 mg/L. All acceptability criteria were considered to be met. The test item was not maintained at ±20% of the initial concentrations therefore the effect levels were estimated based on geometric mean measured concentrations taken from samples at the highest test item nominal concentration. The test item induced no visible effects in carp at any of the concentrations tested including a concentration (nominal 3.2 mg/L) exceeding the water solubility limit. Under the conditions of this study, the 96h LC50 was > 1.9 (C.I. - ) mg/L based on geometric mean measured average concentrations.

Description of key information

LC50 (fish) : > 1.9 (C.I. – ) mg/L or greater than solubility limit, based on geometric mean measured average concentrations, 96hour, freshwater, OECD TG 203, 2004

Toxicity above solubility limit in the test medium

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1.9 mg/L

Additional information

Key study : OECD TG 201, 2004 : The acute toxicity of the test item to Common Carp (Cyprinus carpio) was determined in a 96 hour semi-static test according to OECD TG 203 and EU Method C.1 guidelines under GLP. Following a preliminary range-finding test, fish were exposed, in groups to an aqueous solution of the test item at 0.5, 0.8, 1.3, 2.0 and 3.2 mg/L nominal concentrations with daily renewal. Samples for analytical confirmation of actual exposure concentrations were taken from freshly prepared solutions at the start and after 72 hours of exposure and from 24-hour old solutions after 24 and 96 hours of exposure. Analysis of the samples taken at the start of the final test showed measured concentrations of 1.2, 1.7 and 1.8 mg/L at the nominal concentrations of 1.3, 2.0 and 3.2 mg/L, respectively. During the remainder of the test analysis was only performed on samples taken from nominally 3.2 mg/L. Results of the 24-our old sample taken after 24 hours of exposure showed a measured concentration of 1.1 mg/L. Measured concentrations during the last renewal interval (t=72-96 hours) showed a measured concentration that decreased from 2.7 to 2.0 mg/L. The average exposure concentration at nominal 3.2 mg/L was calculated to correspond to 1.9 mg/L. Analytical results were in agreement with the results obtained during the range-finding test. This confirmed the maximum solubility in ISO-medium was at approximately 2.0 mg/L. All acceptability criteria were considered to be met. The test item was not maintained at ±20% of the initial concentrations therefore the effect levels were estimated based on geometric mean measured concentrations taken from samples at the highest test item nominal concentration. The test item induced no visible effects in carp at any of the concentrations tested including a concentration (nominal 3.2 mg/L) exceeding the water solubility limit. Under the conditions of this study, the 96h LC50 was > 1.9 (C.I. - ) mg/L based on geometric mean measured average concentrations.