Fatty acids, C8-10-(even numberd), Esters with pentaerythritol and adipic acid

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

02 - 24 Aug 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Version / remarks:

adopted in 2008

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Treatment, control
- Sampling method: Samples were taken from each new treatment WAF and the corresponding old solutions (three individual replicates) to determine the WAF concentration stability. The analysis was conducted at 4 different time points for both the new and old solutions during the duration of the test. Samples were collected in duplicate in 30 mL vials with PTFE lined screw cap lids with no headspace.
- Sample storage conditions before analysis: Samples were all analyzed on the day of collection so refrigeration was not necessary.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A water-accommodated fraction (WAF) was tested. The WAF was prepared by adding the appropriate amount of the test substance to the dilution water in glass aspirator bottles using a plastic syringes. The syringes were weighed with and without the test substance to determine the actualloading rate. The vessels were sealed with foil-covered rubber stoppers. The solutions were mixed with Teflon®-coated stirbars on magnetic stirplates. The vortex was set at <10% (actual vortex = 7%) of the static liquid depth. The solutions were mixed for 24 hours (± 1 hour). The solution appeared clear, with bubbled test substance on the surface of the treatment during mixing and after mixing was stopped. At the end of mixing, the solutions were allowed to settle for 1 hour (± 15 minutes). At the end of the settling period, the WAFs were removed from the mixing vessel through the outlet at the bottom of the vessel and placed into 2 L volumetric flasks. 50 µL of Vita chem fresh formula was added to each flask and the test solutions were mixed for an additional 15 minutes prior to the distribution of the WAFs to the test chambers. New WAF solutions were prepared every 48 hours for the renewals.
- Evidence of undissolved material: No undissolved test substance was observed in any of the test chambers.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: Organisms were cultured in the Environmental Toxicology Laboratory, Annandale, USA. The original daphnid culture was supplied by Aquatic Biosystems Inc., Fort Collins, USA.
- Age of parental stock: < 24 h old, taken from 13-day old parents
- Feeding during test
- Food type: Vita-chem fresh water formula (Boyd Enterprises Inc.) and suspension of Pseudokirchneriella subcapitata (Aquatic Biosystems Inc.)
- Amount: Vita-chem fresh water formula: 25 µL/L; Suspension of Pseudokirchneriella subcapitata: 1.3E+08 cells/mL suspension
- Frequency: Vita-chem fresh water formula: Vita-chem feed was added to the treatment WAF before start of the test; Suspension of Pseudokirchneriella subcapitata: 0.35 mL of the suspension were fed at inition of the test and during renewals. Beginning on Day 7, they were fed an additional 0.20 mL suspension on non-renewal days.

ACCLIMATION
- Acclimation conditions: Eight to ten daphnids were kept in 1 L glass culture beakers with approximately 800 mL of reconstituted water. The culture chamber was maintained at 20 ± 1°C under a 16 hour light 8 hour dark photoperiod (10 - 20 foot/candles, 108 - 215 Lux). Two sets of Day 0 cultures were started at least five days a week. Cultures were transferred every other day, with exceptions on holidays or weekends when staff was not present, the brood stock health was evaluated and any mortality, production of males or ephippia was documented as well as any mitigation procedures.
- Type and amount of food: Pseudokirchneriella subcapitata (approximately 4.5 - 6.0E+5 cells/mL). They were also fed 25 µL/L of Vita chem fresh formula mixed on a magnetic stir plate with the reconstituted water prior to feeding with algae.
- Feeding frequency: The culture was fed every other day or as needed based on observed algal clearing.
- Health during acclimation (any mortality observed): The neonates had <10% daily mortality on the experimental start day.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Hardness:

170 - 178 mg/L as CaCO3

Test temperature:

20.4 - 22.4 °C

pH:

7.63 - 8.45

Dissolved oxygen:

6.69 - 8.88 mg/L

Nominal and measured concentrations:

Nominal: 135 mg/L loading rate WAF

Details on test conditions:

TEST SYSTEM
- Test vessel: bottles sealed with PTFE-lined caps
- Type: closed
- Material: glass; Size: 130 mL; Fill volume: 130 mL
- Aeration: dilution water was aerated prior to use
- Renewal rate of test solutio: The treatment and the control were renewed at approx. 48 h intervals using the WAF that were prepared approx. 24 h prior to renewal.
- No. of organisms per vessel: 1
- No. of vessels per concentration: 10
- No. of vessels per control: 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: UV-sterilized deionized well water that is treated and distributed throughout the tesing facility via PVC and stainless steel pipes
- Total organic carbon: < 0.05 ppm
- Alkalinity: 117 mg/L as CaCO3
- Intervals of water quality measurement: Water quality measurements (pH, dissolved oxygen, hardness and temperature) were performed on a sub-sample of new WAF, and on old WAF composited from the replicates in each treatment and control twice weekly during the exposure period.

OTHER TEST CONDITIONS
- Photoperiod: 16 hlight: 8 h dark
- Light intensity: Daylight intensity ranged from approximately 131.2 to 169.8 lux during full daylight.

EFFECT PARAMETERS MEASURED: The daphnids were observed daily for immobilization, reproduction and abnormal behavior/appearance (body length) and the number of young were observed daily, but counted upon renewal.

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 135 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(WAF)

Basis for effect:

reproduction

Details on results:

- Mortality of parent animals: No mortality observed.
- No. of offspring produced per female: control: 112; treatment: 107
- Body length and weight of parent animals: There were no effects on growth, based on the average adult length measurement of 4.9 mm for the control and 5.0 mm for the treatment.
- Time to first brood release or time to hatch: control: 8 -10 days; treatment: 9 - 10 days

Reported statistics and error estimates:

The NOELR value for reproduction was determined using the Two-sample T-Test method.

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: Results of TOC analysis [mg/L]

	New solutions			Old solutions
Test day	control	treatment	Test day	control	treatment
0	0.3005	0.4490	2	0.5506	1.0608
8	0.3127	0.3668	10	0.6724	0.4915
14	0.3976	0.4779	16	0.6451	0.7693
20	0.4448	0.5395	21	0.4143	0.4940

Table 2: Survival and reproduction of daphnids during the test in the control

Test day	Number of live offspring released per replicate exposure vessel										Cumulative daphnids immobilized	Percent survival
	1	2	3	4	5	6	7	8	9	10
1	0	0	0	0	0	0	0	0	0	0	0	100
2	0	0	0	0	0	0	0	0	0	0	0	100
3	0	0	0	0	0	0	0	0	0	0	0	100
4	0	0	0	0	0	0	0	0	0	0	0	100
5	0	0	0	0	0	0	0	0	0	0	0	100
6	0	0	0	0	0	0	0	0	0	0	0	100
7	0	0	0	0	0	0	0	0	0	0	0	100
8	9	0	0	0	0	0	0	0	0	0	0	100
9	0	0	0	0	0	0	0	0	0	0	0	100
10	14	12	14	8	10	13	16	13	4	15	0	100
11	0	0	0	0	0	0	0	0	0	0	0	100
12	34	0	0	0	17	0	0	0	0	0	0	100
13	0	NC	NC	NC	0	NC	NC	NC	NC	NC	0	100
14	0	26	34	18	0	32	19	27	5	29	0	100
15	NC	0	0	0	NC	0	0	0	0	0	0	100
16	50	40	47	42	54	39	39	0	7	41	0	100
17	0	0	0	0	0	0	0	NC	NC	0	0	100
18	54	0	0	0	0	0	0	39	12	0	0	100
19	0	NC	NC	NC	NC	0	NC	0	0	NC	0	100
20	1	28	23	23	41	28	31	19	55	28	0	100
21	7	0	0	0	0	0	0	0	0	0	0	100

NC: neonates present but not counted

Table 3: Survival and reproduction of daphnids during the test in the treatment

Test day	Number of live offspring released per replicate exposure vessel										Cumulative daphnids immobilized	Percent survival
	1	2	3	4	5	6	7	8	9	10
1	0	0	0	0	0	0	0	0	0	0	0	100
2	0	0	0	0	0	0	0	0	0	0	0	100
3	0	0	0	0	0	0	0	0	0	0	0	100
4	0	0	0	0	0	0	0	0	0	0	0	100
5	0	0	0	0	0	0	0	0	0	0	0	100
6	0	0	0	0°	0	*	0	0	0	0	1	90
7	0	0	0	0”	0	*	0	0	0	0	1	90
8	0	0	0	*	0	*	0	0	0	0	2	80
9	0	NC	0	*	0	*	NC	0	NC	0	2	80
10	0	8	13	*	16	*	10	7	15	0	2	80
11	0	0	0	*	0	*	0	0	0	0	2	80
12	17	27	24	*	0	*	21	20	0	0	2	80
13	0	0	NC	*	NC	*	0	0	NC	0	2	80
14	0	0	9	*	30	*	0	0	32	28	2	80
15	NC	NC	0	*	0	*	NC	NC	0	0	2	80
16	46	38	44	*	45	*	38	46	43	0	2	80
17	0	0	0	*	0	*	0	0	0	NC	2	80
18	41	34	2	*	1	*	33	0	0	23	2	80
19	NC	NC	NC	*	NC	*	0	NC	NC	0	2	80
20	2	9	31	*	15	*	0	38	17	9	2	80
21	0	0	0	*	0	*	26	0	0	1	2	80

NC: neonates present but not counted

*: adult immobilized

0°: off-color, 0": off color and small

Reference 2

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline studywithout analytics

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: water accommodated fraction (WAF) of the test item was used. The appropriate method of dosing the test item as a WAF is not covered by the OECD methods; therefore the WAF was prepared with modifications in accordance with the method of Schäfers et al. 2009.
A concentrated stock solution of the test item was prepared by weighing 2 mg of the test item onto a small Teflon plate. The Teflon plate was placed inside a sterile 2 L brown glass Schott bottle with drain port. Dilution water was then added to a volume of 2 L and the solution stirred at 300 rpm for 24 hours at room temperature (21 ± 2°C). The WAF solution was then left to settle for 24 hours before the first 100 mL was drained. The remaining solution was then also drained and used to prepare the test concentration

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: The stock culture was obtained from the German Federal Environment Agency, Institute for Water, Soil, and Air Hygiene (Berlin). Specimens used in the test were bred in the laboratory at Fraunhofer IME
- Age of parental stock: 4- 25 h
- Feeding during test: yes
- Food type: suspensions of living cells of the unicellular alga Desmodesmus subspicatus
- Amount: 0.2 mg carbon
- Frequency: daily (at each media renewal)

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: To maintain the stock culture, batches of 30 - 50 adult daphnids were held at room temperature in approximately 1.8 L of dilution water for one week. During this week, the daphnids were fed daily with an algal suspension (Desmodesmus subspicatus containing Artemio Fluid JBL). Algae growing in the log-phase were centrifuged and the pellet was re-suspended in a minimal volume. Thirty millilitres of this algal suspension was added to 1 L of medium. The medium was changed once per week.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Hardness:

1.0 mmol/L

Test temperature:

18.5 - 19.9 °C

pH:

7.7 - 9.5

Dissolved oxygen:

5.8 - 9.2 mg/L

Nominal and measured concentrations:

Nominal WAF loading rate 1 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, nominal volume, fill volume: glass flasks, 100mL, 100 mL
- Renewal rate of test solution: daily
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water is sourced from the Schmallenberg district water production plants, mostly fed by small springs and percolation. The purification process occurs on site at Fraunhofer IME and includes filtration with activated charcoal, passage through a lime-stone column, and aeration to the point of oxygen saturation. To avoid copper contamination, plastic water pipes are used in the test facilities.
- Chlorine: < 0.02 mg/L
- Alkalinity: 1.3 - 1.6 mmol/L
- Calcium: 0.7 mmol/L
- Magnesium: 0.3 mmol/L
- Conductivity: 203.7 - 216.8 µS/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: daily

OTHER TEST CONDITIONS
- Photoperiod: 16 h light and 8 h darkness
- Light intensity: 501 - 998 lx

EFFECT PARAMETERS MEASURED:
- Number of juvenile, mortality, abnormalities in appearance, behaviour, condition (including male sex) or presence of winter eggs (ephippium) per replicate was recorded daily from day 6 onwards
- Body length (excluding the anal spine) of the adults at study termination

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

other: NOEL

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

other: NOEL

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

(body length)

Remarks on result:

other: adult daphnids

Duration:

21 d

Dose descriptor:

other: NOEL

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

immobilisation

Remarks on result:

other: adult daphnids

Details on results:

- Mortality of parent animals: none
- Cumulative No. of offspring produced per female at the end of exposure: 65 ± 12.4 in the control; 67 ± 8.7 in the 1.0 mg/L treatment loading group
- Body length of parent animals (Day 21): 3.98 ± 0.26 mm in the control; 3.60 ± 1.30 mm in the 1.0 mg/L treatment loading group
- Type and number of morphological abnormalities: none
- Type and number of behavioural abnormalities: none
- Time to first brood release: 10.2 ± 0.95 days in the control; 10.3 ± 1.09 days in the 1.0 mg/L treatment loading group

Reference 3

Endpoint:

long-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 for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

21 d

Dose descriptor:

other: NOEL

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: RA-A, CAS 95912-89-3, Oleon, 2013, RL 2

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 135 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(WAF)

Basis for effect:

reproduction

Remarks on result:

other: RA-A, CAS 71010-76-9, Exxon, 2012, RL1

Conclusions:

The key value for the long-term toxicity of the target substance is a NOEL (21 d) ≥ 1 mg/L (nominal) based on experimental studies for two structurally and chemically closely related source substances (OECD 211, D. magna).

Additional information

There is no study available, in which the long-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 fish. A detailed read-across justification in provided in IUCLID section 13.

The assessment of long-term toxicity to aquatic invertebrates is based on two valid, reliable studies investigating the toxicity of the source substances 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) according to GLP and OECD guideline 211 using Daphnia magna as test organism.

One long term study investigating the toxicity of Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane (CAS-No 95912-89-3) was conducted as a semi-static limit test. A nominal water-accommodated-fraction (WAF) loading rate of 1 mg/L was tested. Since no effects were observed, the nominal NOEL (21 d) value for reproduction, survival and body length of the parent animals was determined to be≥1 mg/L (WAF). Based on this result it can be concluded that no toxicological long-term effects on aquatic invertebrates are expected up to the limit of water solubility for Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane (CAS 95912-89-3).

The second study was conducted under semi-static conditions investigating the toxicity of Decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS 71010-76-9). A loading rate of 135 mg/L (nominal) was applied in the limit test. The WAF solution was prepared by adding the appropriate amount of test substance with subsequent stirring and sampling of the aqueous portions (WAFs) through the outlet at the bottom of the vessels. No significant effect on reproduction was observed in the treatment throughout the test period of 21 d. Hence, the 21 d-NOELR is determined to be >= 135 mg/L on the basis of 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 chronically toxic to aquatic invertebrates.