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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
8 Nov - 30 Nov 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Individual treatments were prepared by adding the appropriate amount of test substance to 2 L of laboratory dilution water in glass aspirator bottles. The test substance was added to the water in the aspirator bottles using stainless steel and glass syringes. The syringes were weighed before and after adding the test substance to determine the actual loading rate. The mixing vessels were closed with foil-covered neoprene stoppers. The mixtures were stirred using a 6.7% (of the static liquid depth) vortex for 23.5 h on magnetic stir plates with Teflon-coated stribars at room temperature. As stirring initiated and after stirring all treatments appeared clear and colourless with the test substance floating at the surface. The mixtures were allowed to settle and equilibrate to test temperature in a waterbath at 17.8 °C for one hour without stirring before removing the aqueous portions (WAFs) through the outlet at the bottom of the vessel for testing.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Source: in-house culture; original culture was supplied by Aquatic Biosystems, Fort Collins, USA on April 11, 2002
- Age at study initiation: < 24 h
- Feeding during test: no

ACCLIMATION
- Acclimation period: Day 0 cultures were started daily using 8 < 24 h old neonates from culture beakers between 12 and 18 days old, exhibiting <20% adult mortality
- Acclimation conditions. same as test
- Type and amount of food: Pseudokirchneriella subcapitata (~ 4.5E5 cells/mL) and 4 mL of a yeast/salmon starter/ Microfeast mixture per 800 mL
- Feeding frequency: daily (five days per week at a minimum)
- Health during acclimation: The pool of neonates selected for the test had ≤ 10% daily mortality on the experimental start day.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
150 mg/L as CaCO3
Test temperature:
20.1 - 20.9
pH:
7.9 - 8.1
Dissolved oxygen:
7.3 - 8.3 mg/L
Nominal and measured concentrations:
Nominal: 62, 132, 251, 503 and 1072 mg/L loading rate WAF
Details on test conditions:
TEST SYSTEM
- Test vessel: Erlenmeyer flask
- Type: closed with ground glass stoppers
- Material: glass, size: 125 mL, fill volume: ~140 mL (no headspace)
- No. of organisms per vessel: 5
- No. of vessels per concentration: 4
- No. of vessels per control: 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The dilution water was prepared from UV-sterilized, deionized well water that was treated and distributed throughout the testing facility via PVC and stainless steel pipes.
- Ca/mg ratio: 1.2:1
- Alkalinity: 96 mg/L as CaCO3
- Conductivity: 500 µmhos
- Culture medium different from test medium: no
- Intervals of water quality measurement: Water quality measurements (pH, dissolved oxygen, temperature) were performed on a sub-sample of the WAFs from each treatment and control on day 0 and on a composite of the replicates in each treatment at termination.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light/ 8 h dark
- Light intensity: Daylight intensity ranged from ~184 - 233 lux during full daylight periods of the study.

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 24 and 48 h test duration. In addition, observations for normal or abnormal behaviour/ appearance were made.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Range finding study: The toxicity of the "water-accomnodated fraction" of the test substance was tested at different loading levels. The WAFs were prepared by adding the appropriate amount of test substance to dilution water in glass aspirator bottles. The vessels were closed and mixed on magnetic stir plates with Teflon coated stirbars for 23 h and 50 minutes. The treatments were allowed to settle for 1 h and five minutes after mixing. Two replicates at each loading level were prepared containing 5 organisms. This study was not conducted according to GLP.
- Test concentrations: 10, 100 and 1000 mg/L loading rate WAFs
- Results used to determine the conditions for the definitive study: No mortality at any loading level was recorded during a exposure time of 48 hours.
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 1 072 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility
Details on results:
- Behavioural abnormalities: none

TOC analysis confirmed the low water solubility of the test substance in water. The total carbon content of the treatment is only slightly higher than that of the control. Thus, the results of the TOC analysis can not be used for the determination of an effect concentration. Thus, the effect concentration is related to the nominal concentration.

Table 1: TOC analysis of the test substance in the test medium

TOC [mg]

Loading rate

0 h

48 h

Control

0.4127

0.4102

62

0.4716

0.4246

132

0.4349

0.4330

251

0.3856

0.3966

503

0.4420

0.4097

1072

0.4415

0.4079

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
17 - 19 Nov 2006
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non-GLP guideline study: Analytical purity is not stated. Age of test organisms is not given. Only two replicates are used. No analytics of the test substance was performed.
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
GLP compliance:
no
Remarks:
But conducted in accordance with ISO/IEC 17025
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Water Accomodated fraction was prepared by moderately stirring 100 mg test substance per litre of medium overnight followed by filtration (Millipore AP 15 glass fiber filter).
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Age at study initiation: not stated
- Length at study initiation: < 2 mm
- Method of breeding: Parental and young Daphnia were held in 40 L glass aquaria, which contain continuously aerated reconstituted water according to ISO 6341 prepared with distilled water. The population were fed once a day (except weekends) with 15 g sera microspan (sera GmbH, 52525 Heinsberg, Germany) homogenized in 1 L deionized water. Every month, 25% of the total volume of the aquaria is replaced by fresh water.
- Feeding during test: no

ACCLIMATION
- Acclimation period: 4 h
- Acclimation conditions: same as test
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Test temperature:
20 ± 0.5 °C
pH:
7.1 - 7.8
Dissolved oxygen:
6.2 - 6.4 mg/L
Nominal and measured concentrations:
Nominal: 100 mg/L loading rate WAF
Details on test conditions:
TEST SYSTEM
- Test vessel: flask
- Material: glass; Size: 100 mL; Fill volume: 50 mL test medium
- Aeration: no
- No. of organisms per vessel: 20
- No. of vessels per concentration: 2
- No. of vessels per control: 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: distilled water
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4:1
- Conductivity: < 1.5 µS/cm
- Culture medium different from test medium: no, reconstituted water according to ISO 6341.
- Intervals of water quality measurement: Temperature was measured in a control flask at the start and at the end of the test. Oxygen and pH were measured in all vessels prior to the addition of Daphnia to the test medium and at the end of the test.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h photoperiod a day
- Light intensity: overhead white fluorescent light

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 24 and 48 h exposure time.

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: limit test
- Range finding study: This study represents a limit test. Therefore, no range finding test was conducted.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility

Table 1: Immobilization of daphnia magna after 24 and 48 h of exposure to the test material

Nominal concentration [mg/L]

Code

Number of individuals per test vessel

Immobile Daphnia [Ind./vessel] 24 h

Immobile Daphnia [Ind./vessel] 48 h

% immobile Daphnia after 24 h of exposure

% immobile Daphnia after 48 h of exposure

Control

A

20

1

2

5

10

B

20

0

2

0

10

C

20

0

1

0

5

100

A

20

1

1

5

5

B

20

0

2

0

10

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
17 - 19 Nov 2006
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non-GLP Guideline study: Analytical purity is not stated. Age of test organisms is not given. Only two replicates are used. No analytics of the test substance performed.
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
GLP compliance:
no
Remarks:
But conducted in accordance with ISO/IEC 17025
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Water Accomodated fraction was prepared by moderately stirring 100 mg test substance per litre of medium overnight followed by filtration (Millipore AP 15 glass fiber filter).
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Age at study initiation: not stated
- Length at study initiation: < 2 mm
- Method of breeding: Parental and young Daphnia were held in 40 L glass aquaria, which contain continuously aerated reconstituted water according to ISO 6341 prepared with distilled water. The population were fed once a day (except weekends) with 15 g sera microspan (sera GmbH, 52525 Heinsberg, Germany) homogenized in 1 L deionized water. Every month, 25% of the total volume of the aquaria is replaced by fresh water.
- Feeding during test: no

ACCLIMATION
- Acclimation period: 4 h
- Acclimation conditions: same as test
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Test temperature:
20 ± 0.5 °C
pH:
7.1 - 7.8
Dissolved oxygen:
6.2 - 6.4 mg/L
Nominal and measured concentrations:
Nominal: 100 mg/L loading rate WAF
Details on test conditions:
TEST SYSTEM
- Test vessel: 100 mL flask
- Material, fill volume: all-glass, 50 mL test medium
- Aeration: No
- No. of organisms per vessel: 20
- No. of vessels per concentration: 2
- No. of vessels per control: 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: distilled water
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4:1
- Conductivity: < 1.5 µS/cm
- Culture medium different from test medium: No, reconstituted water according to ISO 6341.
- Intervals of water quality measurement: Temperature was measured in a control flask at the start and at the end of the test. Oxygen and pH were measured in all vessels prior to the addition of Daphnia to the test medium and at the end of the test.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h photoperiod a day
- Light intensity: overhead white fluorescent light.

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 24 and 48 h exposure time.

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: limit test
- Range finding study: This study represents a limit test. Therefore, no range finding test was conducted.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility

Table 1: Immobilisation of Daphnia magna after 24 and 48 h of exposure to the test material

Nominal concentration [mg/L]

Code

Number of individuals per test vessel

Immobile Daphnia [Ind./vessel] 24 h

Immobile Daphnia [Ind./vessel] 48 h

% immobile Daphnia after 24 h of exposure

% immobile Daphnia after 48 h of exposure

Control

A

20

1

2

5

10

B

20

0

2

0

10

C

20

0

1

0

5

100

A

20

0

1

0

5

B

20

0

2

0

10

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
13 - 15 Dec 2006
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non-GLP guideline study: Analytical purity is not stated. Age of test organisms is not given. Only two replicates are used. No analytics of the test substance was performed.
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
GLP compliance:
no
Remarks:
But conducted in accordance with ISO/IEC 17025
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Water Accomodated fraction was prepared by moderately stirring 100 mg test substance per litre of medium overnight followed by filtration (Millipore AP 15 glass fiber filter).
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Age at study initiation: not stated
- Length at study initiation: < 2 mm
- Method of breeding: Parental and young Daphnia were held in 40 L glass aquaria, which contain continuously aerated reconstituted water according to ISO 6341 prepared with distilled water. The population were fed once a day (except weekends) with 15 g sera microspan (sera GmbH, 52525 Heinsberg, Germany) homogenized in 1 L deionized water. Every month, 25% of the total volume of the aquaria is replaced by fresh water.
- Feeding during test: no

ACCLIMATION
- Acclimation period: 4 h
- Acclimation conditions: same as test
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Test temperature:
20 ± 0.5 °C
pH:
7.3 - 7.6
Dissolved oxygen:
5.6 - 6.0 mg/L
Nominal and measured concentrations:
Nominal: 100 mg/L loading rate WAF
Details on test conditions:
TEST SYSTEM
- Test vessel: flask
- Material:glass; Size: 100 mL; Fill volume: 50 mL test medium
- Aeration: No
- No. of organisms per vessel: 20
- No. of vessels per concentration: 2
- No. of vessels per control: 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: distilled water
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4:1
- Conductivity: < 1.5 µS/cm
- Culture medium different from test medium: No, reconstituted water according to ISO 6341.
- Intervals of water quality measurement: Temperature was measured in a control flask at the start and at the end of the test. Oxygen and pH were measured in all vessels prior to the addition of Daphnia to the test medium and at the end of the test.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h photoperiod a day
- Light intensity: overhead white fluorescent light.

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 24 and 48 h exposure time.

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: limit test
- Range finding study: This study represents a limit test. Therefore, no range finding test was conducted.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility

Table 1: Immobilisation of Daphnia magna after 24 and 48 h of exposure to the test material

Nominal concentration [mg/L]

Code

Number of individuals per test vessel

Immobile Daphnia [Ind./vessel] 24 h

Immobile Daphnia [Ind./vessel] 48 h

% immobile Daphnia after 24 h of exposure

% immobile Daphnia after 48 h of exposure

Control

A

20

0

0

0

0

B

20

0

0

0

0

C

20

0

0

0

0

100

A

20

0

1

0

5

B

20

0

0

0

0

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Remarks:
Summary of available data used for the endpoint assessment of the target substance
Adequacy of study:
key study
Justification for type of information:
Please refer to the analogue justification attached to IUCLID section 13.
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility
Remarks on result:
other: RA-A, CAS 95912-89-3, Oleon, 2006a
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility
Remarks on result:
other:
Remarks:
RA-A, CAS 95912-89-3, Oleon, 2006b
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility
Remarks on result:
other:
Remarks:
RA-A, CAS 95912-89-3, Oleon, 2007
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 1 072 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility
Remarks on result:
other:
Remarks:
RA-A, CAS 71010-76-9, Exxon, 2006
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility
Remarks on result:
other: supporting RA-A, CAS 95912-89-3, Nyco, 2008
Conclusions:
The key value for the short-term toxicity of the target substance is a EL50 (48 h) > 100 mg/L (nominal) based on experimental studies for two structurally and chemically closely related source substances (OECD 202, D. magna).
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
12 - 14 Feb 2008
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non-GLP Guideline study: The test substance was not analytically monitored and no purity of the test substance is given.
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
no
Remarks:
But conducted in accordance with ISO/IEC 17025
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Aqueous solution is saturated with the test substance by dynamic contact setting with a known test substance amount. The preparation consists of 68 h of gentle agitation by magnetic stirring (30 rpm - 60 rpm), followed by 4 h of phase separation. The aqueous phase, containing the soluble substances (Water Accomodated Fraction) is taken to perform the test.
- Differential loading: The test solution is identified according to its loading ratio (massic proportions of substance to be tested/ test medium introduced into the diphasic reactor) while operating the WAF. The implemented loaded ratio is: 1.031/1000.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Source: Breeding of " BfB Oil research Laboratory". They are obtained by acyclic parthenogenesis from at least the third generation.
- Age at study initiation: < 24 h
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
200 - 250 mg/L as CaCo3
Test temperature:
20 ± 1 °C
pH:
7.8 ± 0.2
Dissolved oxygen:
> 90% saturation
Nominal and measured concentrations:
Nominal: 1000 mg/L loading rate WAF
Details on test conditions:
TEST SYSTEM
- Test vessel: test tubes
- Fill volume: 100 mL
- No. of organisms per vessel: 10
- No. of vessels per concentration: 2
- No. of vessels per control: 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: prepared as recommended in the guideline
- Culture medium different from test medium: no
- Intervals of water quality measurement: Temperature was continuously measured. Oxygen content and pH were measured at the beginning and at the end of the test.

OTHER TEST CONDITIONS
- Photoperiod: tested in darkness

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 48 h test duration.
Reference substance (positive control):
not specified
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
mobility
Details on results:
No immobilisation and no behavioural abnormalities were observed after 48 h exposure time.

Description of key information

Key value for chemical safety assessment

Additional information

There is no study available, in which the short-term toxicity of Fatty acids, C8-10-(even numbered), esters with pentaerythritol and adipic acid (EC 948-383-1) to aquatic invertebrates was assessed. Therefore, read-across to the structurally and chemically closely related source substance Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane (CAS 95912-89-3) and Decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS 71010-76-9) was performed in accordance with Regulation (EC) No 1907/2006 Annex XI, section 1.5 in order to fulfill the data requirements for this substance. The source substance is characterized by similar chemical structure and is therefore considered a suitable representative for the assessment of the short-term toxicity of the target substance to aquatic invertebrates. A detailed read-across justification in provided in IUCLID section 13.

The assessment of short-term toxicity to aquatic invertebrates is based on five valid, reliable studies conducted according to OECD guideline 202 investigating the toxicity of the source substances Fatty acids, C8-10, mixed esters with adipic acid and trimethylol-propane (CAS 95912-89-3) and Decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS 71010-76-9).

Four experimental studies investigating the short-term toxicity of Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane (CAS 95912-89-3) to aquatic invertebrates are available. The three key studies were conducted under static conditions using Daphnia magna as test organism. A 100 mg/L loading rate Water Accommodated Fraction (WAF) was tested in a limit test. In all of the three studies the EL50 (48 h) was determined to be > 100 mg/L loading rate based on the nominal concentration. These results are supported by an additional study, which was conducted under static conditions according to OECD guideline 202 using Daphnia magna as test organism. A 1000 mg/L loading rate WAF was used in a limit test. No immobilisation and no behavioural abnormalities were observed in the control and the treatment after 48 h exposure time. Hence, the EL50 (48 h) is determined to be > 1000 mg/L loading rate based on the nominal concentration. Based on this result it can be concluded that no toxicological short-term effects on aquatic invertebrates are to be expected up to the limit of water solubility for Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane (CAS 95912-89-3).

One study is available investigating the toxicity of decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS 71010-76-9) to aquatic invertebrates according to OECD 202 using Daphnia magna as test organism under static conditions. The WAFs were prepared by adding the appropriate amount of test substance with subsequent stirring and sampling of the aqueous portions through the outlet at the bottom of the vessels. Loading rate WAFs of 62, 132, 251, 503 and 1072 mg/L were tested. No immobilisation was observed in any treatment and the control throughout the test period of 48 h. Hence, the 48 h-EL50 was determined to be > 1072 mg/L based on the nominal test concentration. Based on the available results from structurally related read-across substances (in accordance with Regulation (EC) No 1907/2006 Annex XI, 1.5) which are characterized by a similar ecotoxicological profile and comparable structure, it can be concluded that the target substance Fatty acids, C8-10-(even numbered), esters with pentaerythritol and adipic acid (EC 948-383-1) is not actuely toxic to aquatic invertebrates.