2-(1-(3',3'-dimethyl-1'-cyclohexyl)ethoxy)-2-methyl propyl propanoate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 15 February 1995 to 22 April 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

This study was carried out according to international OECD Guideline No. 203 with GLP statement. All validity criteria were fulfilled and no deviations were observed.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

1994-01-31

Specific details on test material used for the study:

- Storage condition of test material: metal bottle at room temperature

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Water samples were taken from the solvent control and all surviving test groups (1.4, 2.5, 4.4, 7.9 and 14 mg/L) at 0 (fresh media), 24, 48, 72 and 96 hours (old media) for quantitative analysis
- Sampling method: An aliquot of each test sample was extracted with three portions (3 x 50 ml) of dichloromethane and each extract filtered through a bed of anhydrous sodium sulphate. The combined extracts were evaporated to dryness and the residue redissolved in ethyl acetate to give a final
theoretical test material concentration of approximately 300 mg/l.
- Sample storage conditions before analysis: no data

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: For the purpose of the main study the test material was prepared using a preliminary solution in 10% v/v Tween 80-dimethylformamide.
Amounts of the test material (0.350. 0.625. 1.10 and 1.975 g) were each separately dissolved in 10% v/v Tween 80-dimethylformamide and
the volume adjusted to 25 ml. An aliquot (2 ml) of these solvent stock solutions were each separately dispersed in approximately 750 ml reverse osmosis water with the aid of ultrasonic disruption. The volume was then adjusted to 1 litre with water prior to dispersal in 20 litres (final volume) of dechlorinated tap water to give the test concentrations of 1.4. 2.5. 4.4 and 7.9 mg/l. To prepare the test concentration of 14 mg/l an amount of the test material (0.28 g) was dissolved in 2 ml of 10% v/v Tween 80-dimethylformamide. This was further dispersed in reverse osmosis water with the aid of ultrasonic disruption. The volume was then adjusted to 1 litre with water prior to dispersal in 20 litres (final volume) of dechlorinated tap water.
- Eluate: solvent
- Differential loading: no data
- Controls: 2 controls (control + solvent control)
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): dimethylformamide
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 10% v/v Tween 80 - dimethylformamide
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no data

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Strain: no data
- Source: Donnington Fish Farm, Upper Swell, Gloucester, U.K.
- Age at study initiation (mean and range, SD): juvenile
- Length at study initiation (length definition, mean, range and SD): The fish had a mean standard length of 4.2 cm (s.d. = 0.1) at the end of the definitive study.
- Weight at study initiation (mean and range, SD): The fish had a mean standard weight of 0.56 g (s.d. = 0.05) at the end of the definitive study.
- Method of breeding: no data
- Feeding during test: no

ACCLIMATION
- Acclimation period: no data
- Acclimation conditions (same as test or not): Fish were maintained in a glass fibre tank with a "single pass" water renewal system. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle (same as test).
- Type and amount of food: The stock fish were fed commercial trout pellets flake food which were discontinued 24 hours prior to the start of the definitive study.
- Feeding frequency: no data
- Health during acclimation (any mortality observed): There were no mortalities in the 7 days prior to the start of the test.

QUARANTINE (wild caught)
Not applicable

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

none

Post exposure observation period:

None

Hardness:

Laboratory tap water dechlorinated was used with a total hardness of approximately 100 mg/L as CaC03 .

Test temperature:

Temperature was maintained at 14 °C throughout the study.

pH:

The pH varied between 7.2 to 7.3 throughout the study.

Dissolved oxygen:

The Dissolved oxygen varied between 8.8 to 10.1 mg O2/L throughout the study.

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal concentrations: 1.4, 2.5, 4.4, 7.9 and 14 mg/L.
Measured concentrations: Analysis of the test preparations showed a marked decline in measured test concentrations after 24 hours and each test media renewal period (see Table 1 in "Any other information on results incl. tables"). Analysis of the test preparations at 0 hours showed the measured concentrations to be near nominal. However analysis of the 24-hour old test media at 24. 48. 72 and 96 hours showed a marked decline in measured concentrations resulting in measured concentrations ranging between 47 - 109% of nominal.

Details on test conditions:

TEST SYSTEM
- Test vessel: glass exposure vessels
- Type (delete if not applicable): covered and sealed
- Material, size, headspace, fill volume: 20 Litre
- Aeration: no auxiliary aeration
- Type of flow-through (e.g. peristaltic or proportional diluter): not applicable
- Renewal rate of test solution (frequency/flow rate): daily renewal
- 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: 0.28 g bodyweight/litre

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Laboratory tap water dechlorinated by passage through an activated carbon filter was used.
- Total organic carbon: no data
- Particulate matter: no data
- Metals: no data
- Pesticides: no data
- Chlorine: free chlorine = 0.18 mg/L ; total chlorine = 0.23 mg/L
- Alkalinity: 104 mg/L
- Ca/mg ratio: no data
- Conductivity: 374
- Culture medium different from test medium: no
- Intervals of water quality measurement: no data

OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: 16 hours light and 8 hours darkness
- Light intensity: no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Any mortalities and adverse reactions to exposure were recorded at 3, 6, 24, 48, 72 and 96 hours after the start of exposure. The criteria of death was taken to be the absence of both respiratory movement and response to physical stimulation.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: average 1.75
- Justification for using less concentrations than requested by guideline: not applicable
- Range finding study: yes (1.4 and 14 mg/L)
- Test concentrations: 1.4, 2.5, 4.4, 7.9 and 14 mg/L.
- Results used to determine the conditions for the definitive study: The results of the range finding study showed no mortalities at the test concentrations of 1.4 mg/l. However, mortalities were observed at 14 mg/l. Additional solubility work was carried out at concentrations in excess of 14 mg/l, however after a period of 24 hours an oily film of test material was observed on the surface of the water thereby indicating 14 mg/l to be the maximum limit of water solubility of the test material under these test conditions.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3.6 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL= 2.7-4.8 mg/L

Duration:

96 h

Dose descriptor:

other: The highest concentration without observed effect

Effect conc.:

1.5 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

In order to give a worst case analysis of the data it is considered justificable to calculate the LC50 value and the highest concentration without observed effect based on the mean 24-hour old test media measured concentrations.

- Behavioural abnormalities: Behavioural responses to exposure (other than death) were observed at test concentrations of greater than 2.5 mg/l. These responses were increased pigmentation, loss of equilibrium and the presence of moribund fish (see Table 3 in "Any other information on results incl. tables").
- Observations on body length and weight: no data
- Other biological observations: none
- Mortality of control: 0%
- Other adverse effects control: none
- Abnormal responses: see above
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The decline in measured test concentrations was observed to be concentration dependant effect ie. a greater loss of test material was observed at the lower test concentrations. This decline in measured test concentrations is considered to be due to adsorption of the test material to the glassware used in the study.
- Effect concentrations exceeding solubility of substance in test medium: none

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

No data

Sublethal observations / clinical signs:

Table 6.1.1/1: Verification of test concentrations

Samples	Nominal concentration (mg/L)	Concentration found (mg/L)	Expressed as a % of the nominal concentration
0 hours	Solvent control 1.4 2.5 4.4 7.9 14.0	< LOQ 1.45 2.65 4.64 8.77 14.2	- 104 106 105 111 102
24 hours	Solvent control 1.4 2.5 4.4 7.9 14.0	< LOQ 0.909 1.65 3.34 8.58 15.1	- 65 66 76 109 108
48 hours	Solvent control 1.4 2.5 4.4	< LOQ 0.743 1.52 3.13	- 53 61 71
72 hours	Solvent control 1.4 2.5 4.4	< LOQ 0.658 1.28 2.92	- 47 51 66
96 hours	Solvent control 1.4 2.5 4.4	< LOQ 0.775 1.47 3.27	- 55 59 74

Table 6.1.1/2: Cumulative mortality data in the definitive study

Nominal concentration (mg/L)	Cumulative mortality (initial population = 10)
	3h	6h	24h	48h	72h	96h
Control	0	0	0	0	0	0
Solvent control	0	0	0	0	0	0
1.4	0	0	0	0	0	0
2.5	0	0	0	0	0	0
4.4	0	0	0	0	2	5
7.9	1	7	10	10	10	10
14.0	8	10	10	10	10	10

Table 6.1.1/3: Behavioural responses to exposure in the definitive study

Nominal concentration (mg/L)	Behavioural responses	Time (hours)
		3	6	24	48	72	96
Control	No abnormalities detected
Solvent control	No abnormalities detected
1.4	No abnormalities detected
2.5	No abnormalities detected
4.4	Increased pigmentation		7/10	8/10	8/10	5/8	5/5
	Loss of equilibrium				2/10
	Moribund					3/8
7.9	Moribund	8/9	3/3
	Loss of equilibrium	1/9		A/D
14.0	Moribund	2/2	A/D

A/D = All fish dead

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test substance to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-hour LC50 value
based on the mean 24-hour old test media measured concentrations of 3.6 mg/l with 95% confidence limits of 2.7 - 4.8 mg/l. The highest concentration without observed effect was 1.5 mg/L.

Executive summary:

This study was performed to assess the acute toxicity of the test material to rainbow trout. The method followed that described in the OECD Guideline No. 203.

Following a preliminary range-finding study, fish were exposed, in groups of 10, to an aqueous dispersion of the test material (with solvent) over a range of concentrations of 1.4, 2.5, 4.4, 7.9 and 14 mg/L for a period of 96 hours under semi-static test conditions. The number of mortalities and any adverse reactions to exposure in each test and control vessel were determined 3 and 6 hours after the start of exposure and then daily throughout the study until termination after 96 hours.

In order to give a worst case analysis of the data it is considered justifiable to calculate the LC50 value and the highest concentration without observed effect based on the mean 24-hour old test media measured concentrations. The 96 hours LC50 for the test material to rainbow trout (Oncorhynchus mykiss) was 3.6 mg/L with 95% confidence limits of 2.7 - 4.8 mg/L. The highest concentration without observed effect was 1.5 mg/L.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Remarks:

Experimental study performed on the C6 isomers of the transformation product of the registered substance.

Adequacy of study:

supporting study

Study period:

October 3, 2013 to October 7, 2013

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

This study was performed with reference to the OECD Guideline 203 under GLP. The test substance is not the registered substance, it's the C6 isomers of the transformation product of the registered substance. This study is considered not assignable as no sufficient information on test conditions is available in the report. Indeed, details of the dissolved oxygen concentration throughout the test, photoperiod, light intensity and validity of the test were not reported in the report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

(minor deviation). The mean total length of test fish (3.2 +/- 0.1 cm) is slightly greater than the recommended one (2.0 +/- 1.0 cm). However, this deviation is not considered to impact the quality of the study.

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

In each test concentration, the test substance was weighed into a 100 mL sample tube and dissolved in water for test heated to approximately 50°C while stirring. The solution was transferred into a 4L test aquarium. Again, water for test heated to approximately 50°C was added to the said sample tube while stirring in order to dissolve the remaining test substance, and then the solution was also transferred into the said test aquarium. The dissolution operation was repeated ten times. After cooling down to room temperature while stirring, test water was prepared by adding water for test so that the total volume was 4L. The test water was analyzed by GC.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Zebra fish (Danio rerio): Lot No. 130904
Supplier: Kamihata Fish Ind., Ltd., 292-1 Chigusa-cho, Hanamigawa-ku, Chiba City, 262-0012 Japan
Date received: September 4, 2013
Mean total length: 3.2 +/- 0.1 cm
Mean bodyweight: 0.31 +/- 0.02 g
Acclimation period: September 4, 2013 to October 3, 2013

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

none

Test temperature:

25.2 +/- 0.2°C

Nominal and measured concentrations:

Nominal concentrations: 6.9, 10.4, 16.2, 22.0 and 36.5 mg/L (+ one control)

Details on test conditions:

TEST SYSTEM
- Test aquarium: Made of glass, amount of test water; 4L.
- No. of organisms per vessel: 10 fish/4L test aquarium

TEST MEDIUM / WATER PARAMETERS
Dechlorinated city water of Saitama.
Analysis of water quality: it was confirmed that water quality was adopted to "the standard of quality of fisheries water".

Reference substance (positive control):

yes

Remarks:

Copper (II) sulfate pentahydrate

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

10.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Observation after 24 and 48h: No mortality was observed in the control and at 6.9 mg/L. 40% mortality was observed at 10.4 mg/L. 100% mortality were observed at 16.2, 22.0 and 36.5 mg/L.
- Observation after 72 and 96h: No mortality was observed in the control and at 6.9 mg/L. 50% mortality was observed at 10.4 mg/L. 100% mortality were observed at 16.2, 22.0 and 36.5 mg/L.

Results with reference substance (positive control):

- Observation after 24h: No mortality was observed at 0.05 mg/L and 30% mortality was observed at 0.5 mg/L.
- Observation after 48h: No mortality was observed at 0.05 mg/L and 60% mortality was observed at 0.5 mg/L.
- Observation after 72 and 96h: No mortality was observed at 0.05 mg/L and 100% mortality was observed at 0.5 mg/L.
The 96 h LC50 value of the reference substance (CuSO4. 5 H2O) for the fish of the same lot is comprised between 0.08 and 0.3 mg/L.

Sublethal observations / clinical signs:

None

Validity criteria fulfilled:

no

Remarks:

The dissolved oxygen concentration throughout the test is not mentioned in the report.

Conclusions:

Under the test conditions, the 96h LC50 for test item was 10.7 mg/L.

Executive summary:

The acute toxicity of the C6 isomers of the transformation product of the registered substance to zebra fish (Danio rerio) was conducted according to OECD Guideline 203 (with one minor deviation) under GLP.

The test substance was tested at concentrations of 6.9, 10.4, 16.2, 22.0 and 36.5 mg/L under semi-static conditions for 96 hours. 10 fish/level were used at the test concentrations and in the control.

After 24 and 48 hours, no mortality was observed in the control and at 6.9 mg/L, 40% mortality was observed at 10.4 mg/L and 100% mortality were observed at 16.2, 22.0 and 36.5 mg/L. After 72 and 96 hours, no mortality was observed in the control and at 6.9 mg/L, 50% mortality was observed at 10.4 mg/L and 100% mortality were observed at 16.2, 22.0 and 36.5 mg/L. Based on this result, the 96h-LC50 was determined to be 10.7 mg/L, based on nominal concentrations.

This study is considered not assignable as no sufficient information on test conditions is available in the report. Indeed, details of the dissolved oxygen concentration throughout the test, photoperiod, light intensity and validity of the test were not reported in the report. In addition, analytical monitoring of the test concentrations were not performed in this study.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

(Q)SAR

Remarks:

QSAR result on the C6 isomers of the transformation product of the registered substance

Adequacy of study:

supporting study

Study period:

Run on 2016-01

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with limited documentation / justification

Remarks:

(Q)SAR method with established scientific validity. The test item, the C6 isomers of the transformation product of the registered substance, falls within the applicability domain of the model.

Justification for type of information:

1. SOFTWARE
EPISUITE v4.1

2. MODEL (incl. version number)
ECOSAR v1.11

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
C6 isomers of the transformation product of the registered substance: C1C(C)(C)CCCC1C(C)OC(C)(C)COH
log Pow: 4.33 (experimental result)

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
Neutral Organics SAR document in "Attached background material".

5. APPLICABILITY DOMAIN
See attached QPRF

6. ADEQUACY OF THE RESULT
See attached QPRF

Qualifier:

according to guideline

Guideline:

other: REACH guidance on QSARs R.6, May 2008

Deviations:

no

Principles of method if other than guideline:

See attached QPRF

GLP compliance:

no

Remarks:

(not relevant)

Specific details on test material used for the study:

- log Pow: 4.33 (experimental result)

Details on sampling:

Not applicable

Details on test solutions:

Not applicable

Test organisms (species):

no data

Details on test organisms:

Not applicable

Test type:

not specified

Water media type:

not specified

Total exposure duration:

96 h

Remarks on exposure duration:

none

Post exposure observation period:

Not applicable

Hardness:

Not applicable

Test temperature:

Not applicable

pH:

Not applicable

Dissolved oxygen:

Not applicable

Salinity:

Not applicable

Nominal and measured concentrations:

Not applicable

Details on test conditions:

Not applicable

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.52 mg/L

Remarks on result:

other: ECOSAR Class: Neutral Organics

Details on results:

Validity of model:
1. Defined Endpoint: Acute toxicity to fish (lethality).
2. Unambigous algorithm: Linear regression QSAR; Log 96h-LC50 (mmol/L) = -0.8981*log Kow + 1.7108. To convert the LC50 from mmol/L to mg/L, multiply by the molecular weight.
3. Applicability domain: applicable to chemicals with log Kow less than 5.0 and molecular weight less than 1000 g/mol. The model can be suitable for chemicals class of neutral organic.
4. Statistical characteristics: N = 296+55 and the coefficient of determination R² = 0.878.
5. Mechanistic interpretation: related to the partitioning of the substance from water into the organisms lipid phase.

Adequacy of prediction: the substance falls within the applicability domain described above and therefore the predicted value can be considered reliable. For more information, see Neutral Organics SAR document in "Attached background material".

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

N = 296+55 and the coefficient of determination R² = 0.878.

Sublethal observations / clinical signs:

No additional information

Validity criteria fulfilled:

yes

Conclusions:

The C6 isomers of the transformation product of the registered substance was predicted to have an 96h-LC50 of 1.52 mg/L to fish after 96h exposure. The substance falls within the applicability domain and therefore the predicted value can be considered reliable.

Executive summary:

The acute toxicity to fish of the C6 isomers of the transformation product of the registered substance was estimated using the ECOSAR v1.11 QSAR model available from the U.S. EPA. The estimation 96h-LC50 was 1.52 mg/L (neutral organics class), calculated from a measured log Kow value (4.33) and a linear regression:

log 96h-LC50 (mmol/L) = -0.8981*log Kow + 1.7108.

The substance falls within the applicability domain and therefore the predicted value can be considered reliable.

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

(Q)SAR

Remarks:

QSAR result on the C6 isomers of the transformation product of the registered substance.

Adequacy of study:

supporting study

Study period:

From 2016-01-05 to 2016-01-06

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification

Remarks:

QSAR value. The test item, the C6 isomers of the transformation product of the registered substance, falls into the applicability domain of the model QSAR.

Justification for type of information:

1. SOFTWARE
iSafeRat® HA-QSAR based on a holistic approach for predicting physicochemical and ecotoxicological endpoints: Short-term toxicity to fish (lethality)

2. MODEL (incl. version number)
iSafeRat® holistic HA-QSAR v1.5

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
C6 isomers of the transformation product of the registered substance: C1C(C)(C)CCCC1C(C)OC(C)(C)COH
Water solubility = 35.0 mg/L (KREATIS internal study, 2015), derived using the log Kow experimental value at 4.33

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
See attached QMRF

5. APPLICABILITY DOMAIN
See attached QPRF

6. ADEQUACY OF THE RESULT
See attached QPRF

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not applicable

Remarks:

(QSAR model)

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

not applicable

Remarks:

(QSAR model)

Principles of method if other than guideline:

The purpose of this QSAR is to accurately predict the acute toxicity to fish as would be expected in a laboratory experiment following OECD Guideline 203 and EC method C.1 for specific, named modes of action (e.g. non-polar narcotics) to provide a value that can effectively replace a 96-hour LC50 value from an experimental study. The regression based method used to achieve this has been fully validated following the OECD (2004) recommendations (refer to the QMRF with JRC/KREATIS QMRF identifier: Q19-46-51-448 for further details).

GLP compliance:

no

Specific details on test material used for the study:

- Mode of action: non-polar narcotic (MOA 1 or MechoA 1.1)
- Log Kow = 4.33 (from experimental study)
- Water solubility = 35.0 mg/L (KREATIS internal study, 2015)

Analytical monitoring:

no

Details on sampling:

Not applicable

Vehicle:

no

Details on test solutions:

Not applicable

Test organisms (species):

other: Danio rerio, Oncorhynchus mykiss, Lepomis macrochirus, Pimephales promelas, Oryzias latipes, Leuciscus idus

Details on test organisms:

Results from the following species were used in the regression: Danio rerio, Oncorhynchus mykiss, Lepomis macrochirus, Pimephales promelas, Oryzias latipes, Leuciscus idus.
Following the principles of Phase Equilibrium Thermodynamics, for narcotic substances, no difference in relationship between solubility and ecotoxicity between fish freshwater species is expected. Any observed differences may be attributed to lifestyle related parameters (e.g. relative differences in storage lipid content between species) and relative duration of study versus bodysize rather than to a specific toxic mechanism causing species differences. In this case, for MOA 1 / MechoA 1.1, no differences were observed in activity based toxicity for the species used.

Test type:

other: QSAR

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

none

Post exposure observation period:

None

Hardness:

The QSAR is based on data from studies performed at acceptable hardness to ensure control survival.

Test temperature:

The temperatures varied from approximately 14 to 25 °C depending on the fish species used to construct the algorithm. While it is recognized that this may contribute to some extent to the variability of the LC50 values found in experimental data, KREATiS has not observed a clear trend suggesting that normalization to temperature would necessarily improve the algorithm (say for trout as opposed to warm water species) for monoconstituents. Nevertheless, this is a recognized area for further research by KREATiS.

pH:

The QSAR is based on data from studies performed with measured pHs between 6.0 - 8.5.

Dissolved oxygen:

The QSAR is based on data from studies performed at acceptable oxygen concentrations (generally >60%).

Salinity:

Not applicable

Nominal and measured concentrations:

The QSAR is based on data from studies performed using measured concentrations or with acceptable stability.

Details on test conditions:

A variety of test designs were accepted: Preferentially results from a semi-static with daily renewal of test solutions and the control or from a flow-through test were used. However, for stable, low volatility substances a static design was accepted (preferably accompanied by analytical measurements over the study period). For suspected volatile substances only tests performed in closed vessels were accepted unless accompanying analytical monitoring proved such a design was not necessary.

Reference substance (positive control):

no

Remarks:

(QSAR model)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 2.0-2.5 mg/L

Details on results:

The predicated value is reliable since the test substance falls within the applicability domain of the model. The value of water solubility value of the test substance is within descriptor domain of the model between log water solubility (in log (mol/L)) of -4.63 to -0.87. Moreover the test substance is attributed to the class of non-polar narcotic compounds.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

95% CL: 2.0-2.5 mg/L
QSAR statistical parameters are given in the QMRF and the QPRF

Sublethal observations / clinical signs:

No additional information

Validity criteria fulfilled:

yes

Remarks:

The substance falls into the applicability domain of the QSAR model.

Conclusions:

The 96-h LC50 of the C6 isomers of the transformation product of the registered substance, based on mortality and measured concentrations, was determined to be 2.2 mg/L with 95%-Confidence Limit between 2.0 and 2.5 mg/L.

Executive summary:

A QSAR prediction was performed to assess the acute toxicity of the test substance to fish. This QSAR has been validated to be compliant with the OECD recommendations for QSAR modelling (OECD, 2004) and predicts the endpoint value which would be expected when testing the substance under experimental conditions in a laboratory following OECD Guideline 203 and EU Method C.1. 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 acute toxicity to fish was determined using a validated QSAR for the Mode of Action in question (MOA 1 or MechoA 1.1, non-polar narcosis). The QSAR is based on validated data for a training set of 67 chemicals derived from 96-hour tests on fish, for which the concentrations of the test substance had been determined by chemical analyses over the test period. The water solubility of the substance given as input was predicted by the iSafeRat® solubility module using the experimental log Kow value of the substance, measured at 4.33. The substance falls within the applicability domain of the model as demonstrated in the QPRF.

The 96-h LC50 of the C6 isomers of the transformation product of the registered substance, based on mortality and measured concentrations, was determined to be 2.2 mg/L with 95%-Confidence Limit between 2.0 and 2.5 mg/L.

Reference 5

Endpoint:

short-term toxicity to fish

Type of information:

(Q)SAR

Remarks:

QSAR result on the C7 isomer of the transformation product of the registered substance

Adequacy of study:

supporting study

Study period:

Run on 2016-03

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with limited documentation / justification

Remarks:

(Q)SAR method with established scientific validity. The substance, the C7 isomer of the transformation product of the registered substance, falls within the applicability domain of the model.

Justification for type of information:

1. SOFTWARE
EPISUITE v4.1

2. MODEL (incl. version number)
ECOSAR v1.11

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
C7 isomer of the transformation product of the registered substance: OCC(C)(C)OC1CC(C)(C)CCCC1C
log Pow: 4.40 (KREATIS QSAR - iSafeRat v1.5, 2016)

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
Neutral Organics SAR document in "Attached background material".

5. APPLICABILITY DOMAIN
See attached QPRF

6. ADEQUACY OF THE RESULT
See attached QPRF

Qualifier:

according to guideline

Guideline:

other: REACH guidance on QSARs R.6, May 2008

Deviations:

no

Principles of method if other than guideline:

See attached QPRF

GLP compliance:

no

Remarks:

(not relevant)

Specific details on test material used for the study:

- log Pow: 4.40 (KREATIS QSAR - iSafeRat v1.5, 2016)

Details on sampling:

Not applicable

Details on test solutions:

Not applicable

Test organisms (species):

no data

Details on test organisms:

Not applicable

Test type:

not specified

Water media type:

not specified

Total exposure duration:

96 h

Remarks on exposure duration:

none

Post exposure observation period:

Not applicable

Hardness:

Not applicable

Test temperature:

Not applicable

pH:

Not applicable

Dissolved oxygen:

Not applicable

Salinity:

Not applicable

Nominal and measured concentrations:

Not applicable

Details on test conditions:

Not applicable

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.31 mg/L

Remarks on result:

other: ECOSAR Class: Neutral Organics

Details on results:

Validity of model:
1. Defined Endpoint: Acute toxicity to fish (lethality).
2. Unambigous algorithm: Linear regression QSAR; Log 96h-LC50 (mmol/L) = -0.8981*log Kow + 1.7108. To convert the LC50 from mmol/L to mg/L, multiply by the molecular weight.
3. Applicability domain: applicable to chemicals with log Kow less than 5.0 and molecular weight less than 1000 g/mol. The model can be suitable for chemicals class of neutral organic.
4. Statistical characteristics: N = 296+55 and the coefficient of determination R² = 0.878.
5. Mechanistic interpretation: related to the partitioning of the substance from water into the organisms lipid phase.

Adequacy of prediction: the substance falls within the applicability domain described above and therefore the predicted value can be considered reliable. For more information, see Neutral Organics SAR document in "Attached background material".

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

N = 296+55 and the coefficient of determination R² = 0.878.

Sublethal observations / clinical signs:

No additional information

Validity criteria fulfilled:

yes

Conclusions:

The C7 isomer of the transformation product of the registered substance was predicted to have an 96h-LC50 of 1.31 mg/L to fish after 96h exposure. The substance falls within the applicability domain and therefore the predicted value can be considered reliable.

Executive summary:

The acute toxicity to fish of the C7 isomer of the transformation product of the registered substance was estimated using the ECOSAR v1.11 QSAR model available from the U.S. EPA. The estimation 96h-LC50 was 1.31 mg/L (neutral organics class), calculated from an estimated log Kow value (4.40) and a linear regression:

log 96h-LC50 (mmol/L) = -0.8981*log Kow + 1.7108.

The substance falls within the applicability domain and therefore the predicted value can be considered reliable.

Reference 6

Endpoint:

short-term toxicity to fish

Type of information:

(Q)SAR

Remarks:

QSAR result on the C7 isomer of the transformation product of the registered substance

Adequacy of study:

supporting study

Study period:

From 2016-03-14 to 2016-03-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification

Remarks:

QSAR value. The substance, the C7 isomer of the transformation product of the registered substance, falls into the applicability domain of the model QSAR.

Justification for type of information:

1. SOFTWARE
iSafeRat® HA-QSAR based on a holistic approach for predicting physicochemical and ecotoxicological endpoints: Short-term toxicity to fish (lethality)

2. MODEL (incl. version number)
iSafeRat® holistic HA-QSAR v1.5

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
C7 isomer of the transformation product of the registered substance: OCC(C)(C)OC1CC(C)(C)CCCC1C
Water solubility = 29.1 mg/L (KREATiS – Internal Study (2016))

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
See attached QMRF

5. APPLICABILITY DOMAIN
See attached QPRF

6. ADEQUACY OF THE RESULT
See attached QPRF

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not applicable

Remarks:

(QSAR model)

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

not applicable

Remarks:

(QSAR model)

Principles of method if other than guideline:

The purpose of this QSAR is to accurately predict the acute toxicity to fish as would be expected in a laboratory experiment following OECD Guideline 203 and EC method C.1 for specific, named modes of action (e.g. non-polar narcotics) to provide a value that can effectively replace a 96-hour LC50 value from an experimental study. The regression based method used to achieve this has been fully validated following the OECD (2004) recommendations (refer to the QMRF with JRC/KREATIS QMRF identifier: Q19-46-51-448 for further details).

GLP compliance:

no

Specific details on test material used for the study:

- Mode of action: non-polar narcotic (MOA 1 or MechoA 1.1)
- log Kow : 4.40 (KREATIS, 2016)
- Water solubility = 29.1 mg/L (KREATIS internal study, 2016)

Analytical monitoring:

no

Details on sampling:

Not applicable

Vehicle:

no

Details on test solutions:

Not applicable

Test organisms (species):

other: Danio rerio, Oncorhynchus mykiss, Lepomis macrochirus, Pimephales promelas, Oryzias latipes, Leuciscus idus

Details on test organisms:

Results from the following species were used in the regression: Danio rerio, Oncorhynchus mykiss, Lepomis macrochirus, Pimephales promelas, Oryzias latipes, Leuciscus idus.
Following the principles of Phase Equilibrium Thermodynamics, for narcotic substances, no difference in relationship between solubility and ecotoxicity between fish freshwater species is expected. Any observed differences may be attributed to lifestyle related parameters (e.g. relative differences in storage lipid content between species) and relative duration of study versus bodysize rather than to a specific toxic mechanism causing species differences. In this case, for MOA 1 or MechoA 1.1, no differences were observed in activity based toxicity for the species used.

Test type:

other: QSAR

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

none

Post exposure observation period:

None

Hardness:

The QSAR is based on data from studies performed at acceptable hardness to ensure control survival.

Test temperature:

The temperatures varied from approximately 14 to 25 °C depending on the fish species used to construct the algorithm. While it is recognized that this may contribute to some extent to the variability of the LC50 values found in experimental data, KREATiS has not observed a clear trend suggesting that normalization to temperature would necessarily improve the algorithm (say for trout as opposed to warm water species) for monoconstituents. Nevertheless, this is a recognized area for further research by KREATiS.

pH:

The QSAR is based on data from studies performed with measured pHs between 6.0 - 8.5.

Dissolved oxygen:

The QSAR is based on data from studies performed at acceptable oxygen concentrations (generally >60%).

Salinity:

Not applicable

Nominal and measured concentrations:

The QSAR is based on data from studies performed using measured concentrations or with acceptable stability.

Details on test conditions:

A variety of test designs were accepted: Preferentially results from a semi-static with daily renewal of test solutions and the control or from a flow-through test were used. However, for stable, low volatility substances a static design was accepted (preferably accompanied by analytical measurements over the study period). For suspected volatile substances only tests performed in closed vessels were accepted unless accompanying analytical monitoring proved such a design was not necessary.

Reference substance (positive control):

no

Remarks:

(QSAR model)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.9 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 1.7-2.2 mg/L

Details on results:

The predicated value is reliable since the test substance falls within the applicability domain of the model. The value of water solubility value of the test substance is within descriptor domain of the model between log water solubility (in log (mol/L)) of -4.63 to -0.87. Moreover the test substance is attributed to the class of non-polar narcotic compounds.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

95% CL: 1.7-2.2 mg/L
QSAR statistical parameters are given in the QMRF and the QPRF

Sublethal observations / clinical signs:

No additional information

Validity criteria fulfilled:

yes

Remarks:

The substance falls into the applicability domain of the QSAR model.

Conclusions:

The 96-h LC50 of the C7 isomer of the transformation product of the registered substance, based on mortality and measured concentrations, was determined to be 1.9 mg/L with 95%-Confidence Limit between 1.7 and 2.2 mg/L.

Executive summary:

A QSAR prediction was performed to assess the acute toxicity of the test substance to fish. This QSAR has been validated to be compliant with the OECD recommendations for QSAR modelling (OECD, 2004) and predicts the endpoint value which would be expected when testing the substance under experimental conditions in a laboratory following OECD Guideline 203 and EU Method C.1. 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 acute toxicity to fish was determined using a validated QSAR for the Mode of Action in question, (MOA 1 or MechoA 1.1, non-polar narcosis). The QSAR is based on validated data for a training set of 67 chemicals derived from 96-hour tests on fish, for which the concentrations of the test substance had been determined by chemical analyses over the test period. The water solubility of the substance given as input was predicted by the iSafeRat® solubility module. The substance falls within the applicability domain of the model as demonstrated in the QPRF.

The 96-h LC50 of the C7 isomer of the transformation product of the registered substance, based on mortality and measured concentrations, was determined to be 1.9 mg/L with 95%-Confidence Limit between 1.7 and 2.2 mg/L.

Description of key information

OECD Guideline 203, GLP, key study, validity 1:

96h-LC50 (Oncorhynchus mykiss) = 3.6 mg/L, based on measured concentrations.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

3.6 mg/L

Additional information

One valid key study, according to the OECD Guideline 203 with GLP standards, is available to assess the acute toxicity of the registered substance on the rainbow trout (Oncorhynchus mykiss). Following a preliminary range-finding study, fish were exposed, in groups of 10, to an aqueous dispersion of the test substance over a range of concentrations of 1.4, 2.5, 4.4, 7.9 and 14 mg/L for a period of 96 hours under semi-static test conditions. The number of mortalities and any adverse reactions to exposure in each test and control vessel were determined 3 and 6 hours after the start of exposure and then daily throughout the study until termination after 96 hours. In order to give a worst case analysis of the data it is considered justifiable to calculate the LC50 value and the highest concentration without observed effect based on the mean 24-hour old test media measured concentrations. Indeed, analysis of the test preparations showed a marked decline in measured test concentrations after 24 hours and each test media renewal period. The 96 hours LC50 was 3.6 mg/L with 95% confidence limits of 2.7 - 4.8 mg/L. The highest concentration without observed effect was 1.5 mg/L.

In addition, the acute toxicity of the transformation product (C6 and C7 isomers) of the registered substance to fish was estimated using two QSAR models and was measured in an experimental study (performed on C6 isomers). The two QSAR models used were ECOSAR v1.11 and iSafeRat holistic approach v1.5. Both QSAR results were considered reliable as the substances, C6 and C7 isomers of the transformation product of the registered substance, fall within the applicability domain of the models. The estimated 96h-LC50 values for C6 isomers were 1.52 mg/L and 2.2 mg/L, using ECOSAR v1.11 and iSafeRat holistic approach v1.5, respectively. The estimated 96h-LC50 values for C7 isomer were 1.31 mg/L and 1.9 mg/L, using ECOSAR v1.11 and iSafeRat holistic approach v1.5, respectively. The experimental 96h-LC50 for C6 isomers, based on nominal concentrations, was 10.7 mg/L, determined in an acute fish study performed on Danio rerio, according to OECD Guideline 203. However, one minor deviation is observed (on the total lenght of test fish) and some information are missing. Therefore, this study is considered not assignable. The worst case endpoint value, estimated at 1.31 mg/L for the C7 isomer, was chosen as key data for the short-term toxicity of the transformation product of the registered substance to fish.

In all cases, these additional data demonstrate that the transformation product (C6 and C7 isomers) of the registered substance is not Toxic (T) in the context of PBT assessment.