2-ethyl-2-[[(1-oxononyl)oxy]methyl]propane-1,3-diyl dinonan-1-oate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

1984

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Test substance concentrations were determined by the Ecotoxicology Analytical Unit.
The concentrations of the substance in the test solutions were measured by a gas chromatography method described below.
Microlitre volumes of a stock solution of the subsrance in anhydrous diethyl ether were pipetted into glass vials and the diethyl ether evaporated to dryness on a sample concentrator at 65 °C under a flow of nitrogen. To each vial a 10 ml volume of Elendt was added, so as to provide calibration standards covering the concentration range 0 - 33 mg/l.To each of these calibration standards, and any 10 ml volumes of samples provided, 2 ml diethyl ether was added and the vials were shaken for two minutes. Sodium chloride (1.5 g) was added to each vial, to separate the layers, and then 500 uI of the upper ether layer was pipetted off and evaporated to dryness on a sample concentrator. The calibration standards and test samples were redissolved in 100 uI of choroform and spiked with 10 uI of an internal standard solution (752mg/l stearic acid methyl ester in chloroform). The spiked extracts were then analysed by gas chromatography with a flame ionisation detector. Results are shown in Table 1.

Vehicle:

no

Details on test solutions:

A 32 mg/I stock solution was prepared by dispersing 0.112 g 0039 (weighed out using the Mettler HK 160 balance in 3.5 Iitres Elendt M7 medium. The stock solution was mixed using a Polytron high shear mixer at 17,000 rpm for approximately 25 minutes.
Test solutions were prepared by adding known volumes of stock to 2000 ml Elendt medium. The test solutions were not renewed at 24 hours. The solutions were prepared 24 hours before the neonates were added to allow adequate time for any unstably dispersed test material to partition out of the water without trapping the Daphnia.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: 1st instar of an in-house laboratory culture
- Age at study initiation (mean and range, SD): < 24 h
- Method of breeding: Cultures were maintained in a synthetic medium with the same chemical composition and similar pH and dissolved oxygen concentration as that used to prepare test solutions. Cultures were maintained in 800 mL of Elendt M7 medium. The cultures were fed daily with cells of the green algae Chlorella vulgaris. Cultures were reset using 10 neonates, when they became gravid their numbers were reduced from 10 to 5. Neonates produced by adults were removed daily.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

233 ± 3 mg/L CaCO3

Test temperature:

19.3 ± 1 °C

pH:

7.5 ± 0.04

Dissolved oxygen:

8.1 ± 0.2 mg/L

Nominal and measured concentrations:

Nominal: 0.0, 1.0, 2.4, 5.6, 13, 32 mg/L
Measured: 0.0, 0.6, 1.2, 2.2, 5.4, 11.5 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Each test vessel (100 mL glass crystallising dish) was submerged in a 2 L crystallising dish containing the appropriate test solution.
- Type: covered to reduce evaporation
- No. of organisms per vessel: 10
- No. of vessels per concentration: 2
- No. of vessels per control: 2

OTHER TEST CONDITIONS
- Photoperiod: 16 h light and 8 h dark

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 24 and 48 h test duration.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 9.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

A number of Daphnia escaped from the submerged test vessels and became trapped at the air/water interface. Since this was considered to be an indirect effect and not a result of direct chemical toxicity, these Daphnia have been excluded from the assessment of effect concentrations. There was no clear relationship between immobilised Daphnia and exposure concentration in the submerged test vessels.

Mean Measured Concentrations (mg/I)	Percentage  Immobile
	24 hrs	48 hrs
0.1	0	5
0.4	0	0
0.9	0	0
1.8	6	6
3.8	0	0
9.3	0	15

Validity criteria fulfilled:

yes

Conclusions:

The 24 and 48 hour EC50s based on mean measured concentrations were greater than the highest concentration tested, ie. >9.3 mg/1.

Executive summary:

The acute toxicity of the substance to the freshwater cladoceran Daphnia magna was determined according to OECO Guideline 202.

The test was conducted under static conditions.

Daphnia were exposed to a nominal range of concentrations 1 - 32 mg/I of the substance. At 0 hours, measured concentrations were about 36 to 60 % of nominal values.

This probably due to the low solubility of the test substance and the difficulties in achieving a stable dispersion. This is also reflected by the fall in all concentrations over the period of the test.

A number of Daphnia escaped from the submerged test vessels and became trapped at the air/water interface. Since this was considered to be an indirect effect and not a result of direct chemical toxicity, these Daphnia have been excluded from the assessment of effect concentrations. There was no clear relationship between immobilised Daphnia and exposure concentration in the submerged test vessels. The 24 and 48 hour EC50s based on mean measured concentrations were greater than the highest concentration tested, ie. >9.3 mg/1.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

other: Read across from analogous substance

Adequacy of study:

supporting study

Study period:

1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

1984

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Test substance concentrations were determined by the Ecotoxicology Analytical Unit.
The concentrations of the substance in the test solutions were measured by a gas chromatography method described below.
Microlitre volumes of a stock solution of the substance in anhydrous diethyl ether were pipetted into 250 ml separating flasks containing 50 ml of Elendt. The diethyl ether was allowed to evaporate for five minutes, so as to provide calibration standards covering the concentration range 0 -14mg/1.To these calibration standards, and each 50ml aliquots of the test samples provided, 5 ml chloroform was added and the flask was shaken for two minutes. The lower chloroform layer was run off and evaporated to dryness at about 65°C under a flow of nitrogen. The standards/samples were redissolved in chloroform and spiked with an internal standard solution of stearic acid methyl ester in chloroform. These were then analysed by gas chromatography with a flame ionisation detector. Results are shown in Table 1.

Vehicle:

no

Details on test solutions:

A 32 mg/I stock solution was prepared by dispersing 0.112 g 0039 (weighed out using the Mettler HK 160 balance in 3.5 Iitres Elendt M7 medium. The stock solution was mixed using a Polytron high shear mixer at 17,000 rpm for approximately 25 minutes.
Test solutions were prepared by adding known volumes of stock to 2000 ml Elendt medium. The test solutions were not renewed at 24 hours. The solutions were prepared 24 hours before the neonates were added to allow adequate time for any unstably dispersed test material to partition out of the water without trapping the Daphnia.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: 1st instar of an in-house laboratory culture
- Age at study initiation (mean and range, SD): < 24 h
- Method of breeding: Cultures were maintained in a synthetic medium with the same chemical composition and similar pH and dissolved oxygen concentration as that used to prepare test solutions. Cultures were maintained in 800 mL of Elendt M7 medium. The cultures were fed daily with cells of the green algae Chlorella vulgaris. Cultures were reset using 10 neonates, when they became gravid their numbers were reduced from 10 to 5. Neonates produced by adults were removed daily.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

233 ± 3 mg/L CaCO3

Test temperature:

19.3 ± 1 °C

pH:

7.5 ± 0.04

Dissolved oxygen:

8.1 ± 0.2 mg/L

Nominal and measured concentrations:

Nominal: 0.0, 1.0, 2.4, 5.6, 13, 32 mg/L
Measured: 0.0, 0.3, 1.2, 2.7, 3.3, 7.6 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Each test vessel (100 mL glass crystallising dish) was submerged in a 2 L crystallising dish containing the appropriate test solution.
- Type: covered to reduce evaporation
- No. of organisms per vessel: 10
- No. of vessels per concentration: 2
- No. of vessels per control: 2

OTHER TEST CONDITIONS
- Photoperiod: 16 h light and 8 h dark

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 24 and 48 h test duration.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 7.6 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

None of the concentrations caused more than 11% immobility and there was no clear relationship between the concentration and the
number of immobilised Daphnia. This suggests that the immobility of Daphnia may have been caused by indirect effects of the test substance or by other unknown
factors.

log Mean Measured Concentrations (mg/I)	Percentage  Immobile
	24 hrs	48 hrs
0	0	0
0.29	5	5
1.2	0	0
2.7	0	0
3.3	0	11
7.6	5	5

Validity criteria fulfilled:

yes

Conclusions:

The 24 and 48 hour EC50s based on the log mean measured concentrations were greater than the highest concentration tested, i.e. >7.6 mg/l.

Executive summary:

The acute toxicity of the substance to the freshwater cladoceran Daphnia magna was determined according to OECD Guideline 202.

The test was conducted under static conditions. Daphnia were exposed to a nominal range of concentrations 1 - 32 mg/l of the substance. The log means of the measured concentrations were in the range 24 to 50 % of nominal values. This was probably due to the low solubility of the test substance, the difficulties in achieving a stable dispersion and because the test solutions were prepared ~4 hours before being analysed.

None of the concentrations of the substance caused more than 11 % immobility and there was no clear relationship between the concentration of the susbtance and number of immobilised Daphnia. This suggests that the immobility of Daphnia may have been caused by indirect effects of test substance or by other unknown factors.

The 24 and 48 hour EC50s based on the log mean measured concentrations were greater than the highest concentration tested, ie. >7.6 mg/l.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

other: Read across from analogous substance

Adequacy of study:

supporting study

Study period:

1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

1984

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

A 100 mg/I stock was prepared by dispersing 0.3 g of the substance in 3 Iitres of hot (- 60 °C) Elendt M7 medium using a Polytron high shear mixer. The stock was mixed for approximately 15 minutes at 15000 rpm, in a 3 litre conical flask to maximise the shearing effect of the blade.
Test media were prepared by adding known volumes of stock to 2000 ml Elendt medium. The test media were not renewed at 24 hours. The media were prepared 24 hours before the neonates were added to allow adequate time for any unstably dispersed test material to partition out of the water without trapping the Daphnia.

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia magna (Crustacea : Cladocera) is a planktonic, filter-feeding invertebrate, widely distributed in the freshwaters of temperate regions around the world. It is widely considered to be sensitive to changes in water quality.
Daphnia magna were cultured in the laboratory according to the procedures described in Ecotoxicology : Cultures were maintained in a synthetic medium with the same chemical composition and similar pH and dissolved oxygen concentration as that used to prepare test media. Cultures were maintained in 800 ml of Elendt M7 medium. The cultures were fed daily with cells of the green algae ChIarella vulgaris.
Cultures were reset using 10 neonates, when they became gravid their numbers were reduced from 10 to 5. Neonates produced by adults were removed daily.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

246 ± 3 mg/L CaCO3

Test temperature:

20.05 ± 0 °C

pH:

7.5 ± 0.1

Dissolved oxygen:

8.1 ± 0.2 mg/L

Nominal and measured concentrations:

Nominal: 0.0, 1.0, 10, 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Each test vessel (100 mL glass crystallising dish) was submerged in a 2 L crystallising dish containing the appropriate test solution.
- Type: covered to reduce evaporation
- No. of organisms per vessel: 10
- No. of vessels per concentration: 2
- No. of vessels per control: 2

OTHER TEST CONDITIONS
- Photoperiod: 16 h light and 8 h dark

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 24 and 48 h test duration.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Nominal Concentrations (mg/I)	24 hrs	48 hrs
0	0	0
1.0	0	5
10	0	35
100	*	30

*Unable to observe Daphnia due to high turbidity of the test medium.

Validity criteria fulfilled:

yes

Conclusions:

The 24 and 48 hour EC50s based on nominal concentrations were greater than the highest concentration tested, i.e. > 100 mg/l. The 48 hour NOEC was 1.0 mg/l.

Executive summary:

The acute toxicity of the substance to the freshwater cladoceran Daphnia magna was determined according to OECD Guideline 202.

The test was conducted under static conditions. Daphnia were not fed during the test.

Daphnia were exposed to a nominal range of concentrations 0, 1.0, 10, and 100 mg/I of the substance dispersed in water. There was no analytical confirmation of these concentrations.

The 24 and 48 hour EC50s based on nominal concentrations were greater than the highest concentration tested, i.e. > 100 mg/l. The 48 hour NOEC was 1.0 mg/l.

Description of key information

Three studies have been conducted (one on the target substance TMP Pelargonate , one on the analogue TMP Triisostearate and one on the analogue TMP, Fatty acids, C14-18 and C18-unsatd., branched and linear, esters ) indicating no effect at the maximum tested concentration.

The target substance is of extremely low water solubility (0.078 µg/L) and therefore the test was performed in supersaturated solution. No lethality / immobility / growth rate reduction could be observed up to the highest attainable test item concentration. Because no hazard was identified, no key values for chemical safety assessment are given below.