Fatty acids, C5-10, esters with pentaerythritol

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:

key study

Study period:

08 - 23 Aug 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study period of 15 days does not meet the criteria of today standard methods recommending 21 days study duration. However, according to the Guidance on information requirements and chemical safety assessment, Chapter R7.b (ECHA, 2012) the study can be regarded as valid for a long-term evaluation as corresponding criteria were fulfilled (e.g. 80% survival in the control, surviving females produce three broods in 21 days or less, > 60 brood neonates per female in the control on average). Thus all requested endpoints could be determined (time to first brood, number of offspring produced per female, growth and survival) and NOELR values for survival or reproductive endpoints could be calculated.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: OECD guideline 202 "Daphnia sp. Acute Immobilization Test and Reproduction Test", Part 2 (1993)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

yes

Remarks:

Only 15 d test duration; Adopted in 1998

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: One set of final and initial WAF samples was collected on day 14 for chemical analysis. WAF samples (initial and final) of the control, the lowest (24 mg/L), middle (242 mg/L), and highest ( 2570 mg/L) were collected in duplicate.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Generation of WAFs was carried out using a modification of the procedure developed by Anderson et al. (1974). Six individual WAF setups, one for each of the five exposure concentrations and one for control were prepared. An appropriate amount of test material was added to 2000 mL of Mobil Technical Center well water in each corresponding WAF aspirator bottle. Bottles were capped with parafilm and stirred for 20 h at room temperature. After stirring, the phases were allowed to separate for 4 h. 800 mL of the water phase were subsequently drained from the bottom spout.
- Differential loading: yes
- Controls: Mobil Technical Center well water without test material.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: The primary culture originated from Aquatic Research Organisms, Hampton, NH. The in-house culture had been maintained in the laboratory since June 19, 1996.
- Age of parental stock: Daphnids used for the test were less than 24 h old.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

15 d

Test temperature:

19.3-20.5 °C

pH:

7.84-8.41

Nominal and measured concentrations:

Nominal: 0, 24, 97, 242, 1018 and 2570 mg/L
Measured: Below the detection limit of 0.03 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 mL polystyrene cups with 40 mL of test solution
- Type (delete if not applicable): covered with a plexiglass sheet
- Renewal rate of test solution (frequency/flow rate): every 48 h
- 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: Mobil Technical Center well water
- Intervals of water quality measurement: at the beginning of the study and after every renewal period

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

EFFECT PARAMETERS MEASURED: immobilisation and reproduction

Reference substance (positive control):

no

Duration:

15 d

Dose descriptor:

EL50

Effect conc.:

> 2 570 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

immobilisation

Duration:

15 d

Dose descriptor:

EL50

Effect conc.:

> 2 570 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

reproduction

Duration:

15 d

Dose descriptor:

NOELR

Effect conc.:

>= 2 570 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

immobilisation

Duration:

15 d

Dose descriptor:

NOELR

Effect conc.:

>= 2 570 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

reproduction

Details on results:

- Mortality of parent animals: 0%

No effects were observed in the range of water solubility with an NOELR = 2570 mg/L (15 d) based on reproduction and immobilisation.

Table 1: Final total reproduction counts taken during the static renewal three-brood chronic toxicity study of the WAF of the test substance to Daphnia magna

Replicates	Control	Exposure Concentrations (ppm)
		24	97	242	1018	2570
A	89	107	102	125	109	112
B	94	113	111	101	135	127
C	115	123	129	120	88	68
D	108	92	92	125	130	109
E	124	122	127	100	123	118
F	117	120	116	121	125	127
G	115	98	0	124	142	129
H	89	79	102	103	128	129
I	114	128	136	102	79	116
J	128	151	140	56	130	116
Totals	1093	1133	1055	1077	1189	1151
Average	109	113	106	108	119	115
Std. Dev.	14	20	40	21	21	18

 

Reference 2

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 - 24 Aug 2011

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study with acceptable restrictions: The presence of neonates in the treatment group was not recorded on day 11.

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

Key result

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

Results with reference substance (positive control):

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 3

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Remarks:

2 substances are available for read across

Adequacy of study:

weight of evidence

Justification for type of information:

see the attached justification in section 13

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

GLP compliance:

yes

Key result

Duration:

15 d

Dose descriptor:

EL50

Effect conc.:

> 2 570 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

immobilisation

Remarks on result:

other: 2,2-bis[(octanoyloxy)methyl]butyl decanoate (CAS 11138-60-6)

Key result

Duration:

15 d

Dose descriptor:

EL50

Effect conc.:

> 2 570 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

reproduction

Remarks on result:

other: 2,2-bis[(octanoyloxy)methyl]butyl decanoate (CAS 11138-60-6)

Key result

Duration:

15 d

Dose descriptor:

NOELR

Effect conc.:

>= 2 570 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

immobilisation

Remarks on result:

other: 2,2-bis[(octanoyloxy)methyl]butyl decanoate (CAS 11138-60-6)

Key result

Duration:

15 d

Dose descriptor:

NOELR

Effect conc.:

>= 2 570 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

reproduction

Remarks on result:

other: 2,2-bis[(octanoyloxy)methyl]butyl decanoate (CAS 11138-60-6)

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

> 135 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test mat. / WAF

Basis for effect:

reproduction

Remarks on result:

other: Decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS 71010-76-9)

Validity criteria fulfilled:

not applicable

Remarks:

read across

Conclusions:

The substance, CAS 68424-31-7 ; EC 270-291-9, is analogous to the substances to be read across to, in terms of basic form, and the degree of substitution of functional groups is not considered to effect the proposed read across for the endpoint of long-term toxicity to aquatic invertebrates. Based on the available information to read across to, the substance is not expected to be toxic upto the limit of solubility in water.

Description of key information

 No toxic effects up to the limit of water solubility. 

Additional information

There is no long-term test on aquatic invertebrates available for pentaerythritol tetraesters of n-decanoic, n-heptanoic, n-octanoic and n-valeric acids (CAS 68424-31-7).Since all substances within this polyol esters category are readily biodegradable, it is not foreseen that they will pose a risk to aquatic invertebrates on the long-term.However, due to the poor water solubility of the test substance and as long term toxicity to aquatic organisms is a mandatory endpoint in accordance to Regulation (EC) No 1907/2006, Annex IX 9.1 Aquatic toxicity, we propose a read across to proposed tests on long term toxicity to invertebrates for the PE esters CAS 68424-31-7 (tetra-esterification products of C5, C7, C8, C10 fatty acids with pentraerythritol), CAS 15834-04-5, CAS 68424-30-6 and CAS 85049-33-8.  CAS 68424-31-7 (C5, C7, C8 and C10) is a highly complex UVCB, with varying chain lengths and combinations of structures. CAS 15834-04-5 has the shortest chain length (C5) of PE esters, it is a mono-constituent and often is a constituent with many of the other polyol esters. Due to its smaller molecular size it is expected to have the highest bioavailability. CAS 68424-30-6 (C5, C7, C9 iso) was chosen on the basis of its isononanoic acid content to evaluate possible adverse effects.CAS 85049-33-8 was selected in order to confirm that no chronic toxicity to aquatic organisms occurs for the polyol esters with medium chain length of the category C8-C16 (even numbered).

Reliable tests on the long-term toxicity to aquatic invertebrates are also provided which indicate that the polyol esters will not exhibit long-term effects to aquatic invertebrates up to the limit of water solubility.

The first study with the read-across substance decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS No. 71010-76-9) was conducted under semi-static conditions according to OECD 211 using Daphnia magna as test organism (Letinski & Bragin, 2011).The test 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. A loading rate WAF of 135 mg/L (nominal) was applied in a limit test. 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.

The second study investigating the long term toxicity to aquatic invertebrates was performed with the read-across substance fatty acids, C8-10 (even numbered), di-and triesters with propylidynetrimethanol (CAS 11138-60-6) according to OECD guideline 202 "Daphnia sp. Acute Immobilization Test and Reproduction Test", Part 2 (1993) under semi-static conditions (Low, 1996). This test guideline is similar to OECD 211 which was adopted in 1998. The study period of 15 days does not meet the criteria of today standard methods recommending 21 days study duration. However, according to the Guidance on information requirements and chemical safety assessment, Chapter R7.b (ECHA, 2012) the study can be regarded as valid for a long-term evaluation as corresponding criteria were fulfilled (e.g. 80% survival in the control, surviving females produce three broods in 21 days or less, > 60 brood neonates per female in the control on average). In addition, all requested endpoints could be determined (time to first brood, number of offspring produced per female, growth and survival) and NOELR values for survival or reproductive endpoints could be calculated. In this study Daphnia magna were exposed to nominal concentrations of 0, 24, 97, 242, 1018 and 2570 mg/L (prepared as water accommodated fraction, WAF). No effects were observed up to the highest test concentration resulting in a NOELR of ≥ 2570 mg/L based on reproduction and immobilization.

As it can be seen in the data matrix of the category justification in section 13 and the overall endpoint summary IUCLID 6.1, all reliable data in the category support the hazard assessment of each category member by showing a consistent pattern of results, i.e. no toxic effects to aquatic organisms were observed up to the limit of water solubility.