Soybean oil, oligomerized

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:

25/03/2014 - 16/04/2014

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Read-across to study according to international guideline (OECD guideline 211) under GLP. Validity citeria met, no deviations from guideline reported.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not relevant

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 1, 10 and 100 mg/L water accomodated fractions (WAF)

There was no analytical verification of test concentrations. However, a verification of the presence of dissolved substance was carried out as part of a non-GLP-pre-test by GC/MS and NMR analysis.
- Sampling method: Three WAF’s were prepared with loadings of 1, 10, 100 mg of VEOPOL per liter water and stirred for 24 h. After a separation phase of 2 h the water fractions were analyzed to look for soluble free fatty acids by GC/MS and NMR. Additionally a control blank was prepared according to the same procedure. Three times 40 mL water samples of each WAF were taken.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test loadings were prepared by placing a small Teflon plate, carrying the test item (200, 20, or 2 mg) on its surface, in a WAF-bottle containing two litres of water. WAFs were prepared by stirring the water for 24 hours at room temperature (about 20°C). The water was stirred slowly to avoid bubble and foam formation using a star shape magnetic stirring bar. After stirring the contents the vessels were left to settle for 2 h. Then the saturated aqueous phase was taken out of the drain port. The first fraction 0-100 mL was discarded. Prior to testing the test media was not stored for more than 1 - 2 hours. Eluates were used for the rinsing (in order to saturate the surfaces) and filling of the test flasks (50 mL volume) for toxicity testing.
- Controls: The control consisted of dilution water only and was prepared in a mixing vessel treated in accordance to the eluate preparation. The control group was conducted with ten replicates with individual specimens each.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): The 100 mg from the highest concentration was not completely solved. A small amount of test item was found on the small Teflon plate after stirring the water for 24 hours. However, no oil film on the eluate or turbity of the test medium could be observed as judged by visual inspection.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: Straus
- Source: German Federal Environment Agency, Institute for Water, Soil, and Air Hygiene (Berlin)
- Age of parental stock (mean and range, SD): less than 24 h old
- Feeding during test
- Food type: Desmodesmus subspicatus
- Amount: 0.2 mg C/Daphnia/day
- Frequency: daily

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: sieving

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

Not relevant

Hardness:

1.1 mmol/L

Test temperature:

19.0 - 21.6ºC

pH:

7.94 - 9.06 (not more than 1.5 increase in any single test vessel)

Dissolved oxygen:

8.40 - 10.70 mg/L

Salinity:

Not relevant

Nominal and measured concentrations:

Nominal: 1, 10 and 100 mg/L water accomodated fraction (WAF)

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: glass containing 50 mL of test medium
- Aeration: none
- 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: Purified local tap water was used in accordance with the OECD guideline [4]. The tap water was sourced from the Schmallenberg district water production plants, mostly fed by small springs and percolation. The purification process occured on site at Fraunhofer IME and included 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 were used in the test facilities.
- Total organic carbon: 0.2638 mg/L
- Chlorine: 0.02 mg/L
- Alkalinity: 1.6 mmol/L
- Conductivity: 223.2 uS/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: The light intensity, temperature, pH, and oxygen saturation were measured daily.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light / 8 h dark
- Light intensity: 698 - 835 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): The numbers of immobile daphnia were visually determined daily. Any abnormalities in appearance and behaviour were recorded. The newborn daphnia per beaker were counted and removed at each water renewal, abnormalities in condition (including male sex) or presence of winter eggs were checked and recorded. At study termination, length of the adults up to the highest treatment without significant mortality was be measured by digital photography and image analysis and compared with the equally measured control animals.

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water accomodated fraction (WAF)

Basis for effect:

mortality

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water accomodated fraction (WAF)

Basis for effect:

growth

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water accomodated fraction (WAF)

Basis for effect:

reproduction

Details on results:

- Mortality of parent animals: 0%
- Body length of parent animals: 4.34 - 4.53 mm
- Type and number of morphological abnormalities: no clinical sub-lethal signs were observed in surviving parental daphnia until the end of the test.
- Type and number of behavioural abnormalities: no clinical sub-lethal signs were observed in surviving parental daphnia until the end of the test.
- Time to first brood release or time to hatch: 13.3 - 14.8 days
- Brood size: 53.7 - 64.6 (cumulative)
- Effect concentrations exceeding solubility of substance in test medium: The 100 mg from the highest concentration was not completely solved. A small amount of test item was found on the small Teflon plate after stirring the water for 24 hours. However, no oil film on the eluate or turbity of the test medium could be observed as judged by visual inspection.

Results with reference substance (positive control):

No data

Reported statistics and error estimates:

The following endpoints observed in the reproduction test were evaluated quantitatively:
• Mortality (immobility) of parental daphnia
• Mean age at day of first brood
• Cumulative number of live offspring per surviving female at day 21 (study end)
• Individual length of adults

The subject of the statistical evaluation was the replicate (n=10). Juveniles from females that died before test end were excluded from the evaluation. The data were tested for variance homogeneity, normal distribution, and outliers. The NOEL and LOEL were calculated by ANOVA followed by Dunnett`s Multiple t-test Procedure. When the test results showed a concentration-response relationship, the data were analysed by regression to determine the EL50 including the 95% confidence interval as well as the EL10 using Probit-analysis assuming log-normal distribution of the values.The NOEL was reported when the statistical test failed to find a difference between the limit treatment and the control. All statistical evaluations were conducted using the computer software ToxRat® Professional v. 2.10.

Survival, growth and reproduction averages per concentration.

Loading concentration	Parental mortality	Adult Growth (length on day 21)	Age at first brood	Cumulative offspring per female
(mg/L)	(%)	Mean± SD (mm)	Mean± SD (days)	Mean± SD (Ind.)
Control	0	4.34 ± 0.11	13.3 ± 1.55	64.6 ± 10.12
1.0	0	4.49 ± 0.27	14.7 ± 0.63	54.0 ± 10.58
10.0	0	4.35 ± 0.21	14.0 ± 1.90	53.7 ± 11.34
100.0	0	4.53 ± 0.26	14.8 ± 1.98	55.8 ± 10.2
NOEL*	≥ 100.0 mg/L	≥ 100.0 mg/L	≥ 100.0 mg/L	≥ 100.0 mg/L

Offspring per test replicate per day.

Time	Replicate	Control	1 mg/L	10 mg/L	100 mg/L
Day 10	1	0	0	0	*
	2	0	0	0	0
	3	0	0	0	0
	4	0	0	0	4
	5	0	0	0	0
	6	0	0	0	0
	7	0	0	0	0
	8	0	0	0	0
	9	0	0	0	0
	10	0	0	0	0
	# Replicates	10	10	10	9
	Mean	0.00	0.00	0.00	0.44
	Std.Dev	0.00	0.00	0.00	1.33
	CV%				300.0
Day 11	1	0	0	0	*
	2	1	0	0	0
	3	0	0	0	0
	4	0	0	0	0
	5	0	0	5	0
	6	1	0	4	0
	7	0	0	0	0
	8	0	0	9	0
	9	5	0	0	0
	10	10	0	0	0
	# Replicates	10	10	10	9
	Mean	1.70	0.00	1.80	0.00
	Std.Dev	3.30	0.00	3.16	0.00
	CV%	194.2		175.3
Day 14	1	5	7	4	*
	2	20	13	15	21
	3	21	11	0	0
	4	20	5	14	7
	5	2	7	1	9
	6	1	9	0	10
	7	2	3	11	0
	8	20	17	2	0
	9	18	7	20	28
	10	0	0	0	20
	# Replicates	10	10	10	9
	Mean	10.90	7.90	6.70	10.56
	Std.Dev	9.49	4.91	7.56	10.32
	CV%	87.1	62.1	112.8	97.8

Day 16	1	20	21	18	*
	2	0	0	0	0
	3	0	26	17	32
	4	22	19	24	27
	5	14	27	21	14
	6	19	23	19	21
	7	20	15	0	33
	8	0	0	26	16
	9	0	22	0	0
	10	24	33	28	0
	# Replicates	10	10	10	9
	Mean	11.90	18.60	15.30	15.89
	Std.Dev	10.55	10.93	11.11	13.52
	CV%	88.6	58.7	72.6	85.1
Day 18	1	0	0	0	*
	2	23	25	24	26
	3	25	0	0	0
	4	0	0	0	0
	5	23	0	0	25
	6	24	0	17	2
	7	20	14	18	13
	8	26	26	2	0
	9	26	0	29	19
	10	27	27	26	29
	# Replicates	10	10	10	9
	Mean	19.40	9.20	11.60	12.67
	Std.Dev	10.42	12.38	12.31	12.41
	CV%	53.7	134.6	106.1	98.0
Day 21	1	22	20	20	*
	2	25	25	27	0
	3	24	24	23	23
	4	23	21	25	27
	5	26	24	18	0
	6	0	17	0	28
	7	29	0	29	15
	8	24	25	19	28
	9	23	27	22	0
	10	11	0	0	25
	# Replicates	10	10	10	9
	Mean	20.70	18.30	18.30	16.22
	Std.Dev	8.64	10.07	10.24	12.78
	CV%	41.7	55.0	56.0	78.8

*Male-daphnid.

Adult length of each replicate at day 21 [mm].

Replicate	Control	1 mg/L	10 mg/L	100 mg/L
1	4.16	4.71	4.49	*
2	4.35	4.14	4.15	4.62
3	4.47	4.9	4.46	4.4
4	4.35	4.55	4.61	4.67
5	4.27	4.65	4.46	4.56
6	4.37	4.52	4.51	4.43
7	4.51	4.39	4.22	4.92
8	4.24	4.74	3.99	4.81
9	4.25	4.1	4.47	4.27
10	4.42	4.23	4.13	4.09
# Replicates	10	10	10	9
Mean	4.34	4.49	4.35	4.53
Std.Dev	0.11	0.27	0.21	0.26
CV%	2.5	6.0	4.8	5.8

Validity criteria fulfilled:

yes

Remarks:

The mortality was 0.0% and mean offspring number was 64.6.

Conclusions:

The chronic (21d-NOELR) toxicity of VEOPOL 212045 (Linseed Standoil 45 poises) towards Daphnia magna with regard to mortality, adult growth and reproduction (time of first brood, number of offspring) is >= 100 mg/L for each of the aforementioned endpoints.

Executive summary:

The chronic toxicity of VEOPOL 212045 (Linseed Standoil 45 poises) towards Daphnia magna was investigated according to OECD guideline 211 under GLP. Daphnids were exposed to water accomodated fractions (WAFs) of 1, 10 and 100 mg/L in a semi-static setup for 21 days, while mortality, adult growth and reproduction (time of first brood, number of offspring) were observed. The chronic toxicity with regard to mortality, adult growth and reproduction was found to be >= 100 mg/L for each of the aforementioned endpoints. The presence of dissolved material in the different WAF preparations was confirmed prior to the study by GC/MS and NMR analysis as part of a non-GLP approach.

Reference 2

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

See attached justification

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water accomodated fraction (WAF)

Basis for effect:

mortality

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water accomodated fraction (WAF)

Basis for effect:

growth

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water accomodated fraction (WAF)

Basis for effect:

reproduction

Details on results:

- Mortality of parent animals: 0%
- Body length of parent animals: 4.34 - 4.53 mm
- Type and number of morphological abnormalities: no clinical sub-lethal signs were observed in surviving parental daphnia until the end of the test.
- Type and number of behavioural abnormalities: no clinical sub-lethal signs were observed in surviving parental daphnia until the end of the test.
- Time to first brood release or time to hatch: 13.3 - 14.8 days
- Brood size: 53.7 - 64.6 (cumulative)
- Effect concentrations exceeding solubility of substance in test medium: The 100 mg from the highest concentration was not completely solved. A small amount of test item was found on the small Teflon plate after stirring the water for 24 hours. However, no oil film on the eluate or turbity of the test medium could be observed as judged by visual inspection.

Results with reference substance (positive control):

No data

Reported statistics and error estimates:

The following endpoints observed in the reproduction test were evaluated quantitatively:
• Mortality (immobility) of parental daphnia
• Mean age at day of first brood
• Cumulative number of live offspring per surviving female at day 21 (study end)
• Individual length of adults

The subject of the statistical evaluation was the replicate (n=10). Juveniles from females that died before test end were excluded from the evaluation. The data were tested for variance homogeneity, normal distribution, and outliers. The NOEL and LOEL were calculated by ANOVA followed by Dunnett`s Multiple t-test Procedure. When the test results showed a concentration-response relationship, the data were analysed by regression to determine the EL50 including the 95% confidence interval as well as the EL10 using Probit-analysis assuming log-normal distribution of the values.The NOEL was reported when the statistical test failed to find a difference between the limit treatment and the control. All statistical evaluations were conducted using the computer software ToxRat® Professional v. 2.10.

Survival, growth and reproduction averages per concentration.

Loading concentration	Parental mortality	Adult Growth (length on day 21)	Age at first brood	Cumulative offspring per female
(mg/L)	(%)	Mean± SD (mm)	Mean± SD (days)	Mean± SD (Ind.)
Control	0	4.34 ± 0.11	13.3 ± 1.55	64.6 ± 10.12
1.0	0	4.49 ± 0.27	14.7 ± 0.63	54.0 ± 10.58
10.0	0	4.35 ± 0.21	14.0 ± 1.90	53.7 ± 11.34
100.0	0	4.53 ± 0.26	14.8 ± 1.98	55.8 ± 10.2
NOEL*	≥ 100.0 mg/L	≥ 100.0 mg/L	≥ 100.0 mg/L	≥ 100.0 mg/L

Offspring per test replicate per day.

Time	Replicate	Control	1 mg/L	10 mg/L	100 mg/L
Day 10	1	0	0	0	*
	2	0	0	0	0
	3	0	0	0	0
	4	0	0	0	4
	5	0	0	0	0
	6	0	0	0	0
	7	0	0	0	0
	8	0	0	0	0
	9	0	0	0	0
	10	0	0	0	0
	# Replicates	10	10	10	9
	Mean	0.00	0.00	0.00	0.44
	Std.Dev	0.00	0.00	0.00	1.33
	CV%				300.0
Day 11	1	0	0	0	*
	2	1	0	0	0
	3	0	0	0	0
	4	0	0	0	0
	5	0	0	5	0
	6	1	0	4	0
	7	0	0	0	0
	8	0	0	9	0
	9	5	0	0	0
	10	10	0	0	0
	# Replicates	10	10	10	9
	Mean	1.70	0.00	1.80	0.00
	Std.Dev	3.30	0.00	3.16	0.00
	CV%	194.2		175.3
Day 14	1	5	7	4	*
	2	20	13	15	21
	3	21	11	0	0
	4	20	5	14	7
	5	2	7	1	9
	6	1	9	0	10
	7	2	3	11	0
	8	20	17	2	0
	9	18	7	20	28
	10	0	0	0	20
	# Replicates	10	10	10	9
	Mean	10.90	7.90	6.70	10.56
	Std.Dev	9.49	4.91	7.56	10.32
	CV%	87.1	62.1	112.8	97.8

Day 16	1	20	21	18	*
	2	0	0	0	0
	3	0	26	17	32
	4	22	19	24	27
	5	14	27	21	14
	6	19	23	19	21
	7	20	15	0	33
	8	0	0	26	16
	9	0	22	0	0
	10	24	33	28	0
	# Replicates	10	10	10	9
	Mean	11.90	18.60	15.30	15.89
	Std.Dev	10.55	10.93	11.11	13.52
	CV%	88.6	58.7	72.6	85.1
Day 18	1	0	0	0	*
	2	23	25	24	26
	3	25	0	0	0
	4	0	0	0	0
	5	23	0	0	25
	6	24	0	17	2
	7	20	14	18	13
	8	26	26	2	0
	9	26	0	29	19
	10	27	27	26	29
	# Replicates	10	10	10	9
	Mean	19.40	9.20	11.60	12.67
	Std.Dev	10.42	12.38	12.31	12.41
	CV%	53.7	134.6	106.1	98.0
Day 21	1	22	20	20	*
	2	25	25	27	0
	3	24	24	23	23
	4	23	21	25	27
	5	26	24	18	0
	6	0	17	0	28
	7	29	0	29	15
	8	24	25	19	28
	9	23	27	22	0
	10	11	0	0	25
	# Replicates	10	10	10	9
	Mean	20.70	18.30	18.30	16.22
	Std.Dev	8.64	10.07	10.24	12.78
	CV%	41.7	55.0	56.0	78.8

*Male-daphnid.

Adult length of each replicate at day 21 [mm].

Replicate	Control	1 mg/L	10 mg/L	100 mg/L
1	4.16	4.71	4.49	*
2	4.35	4.14	4.15	4.62
3	4.47	4.9	4.46	4.4
4	4.35	4.55	4.61	4.67
5	4.27	4.65	4.46	4.56
6	4.37	4.52	4.51	4.43
7	4.51	4.39	4.22	4.92
8	4.24	4.74	3.99	4.81
9	4.25	4.1	4.47	4.27
10	4.42	4.23	4.13	4.09
# Replicates	10	10	10	9
Mean	4.34	4.49	4.35	4.53
Std.Dev	0.11	0.27	0.21	0.26
CV%	2.5	6.0	4.8	5.8

Validity criteria fulfilled:

yes

Remarks:

The mortality was 0.0% and mean offspring number was 64.6.

Conclusions:

The chronic (21d-NOELR) toxicity of VEOPOL 212045 (Linseed Standoil 45 poises) towards Daphnia magna with regard to mortality, adult growth and reproduction (time of first brood, number of offspring) is >= 100 mg/L for each of the aforementioned endpoints.

Executive summary:

The chronic toxicity of VEOPOL 212045 (Linseed Standoil 45 poises) towards Daphnia magna was investigated according to OECD guideline 211 under GLP. Daphnids were exposed to water accomodated fractions (WAFs) of 1, 10 and 100 mg/L in a semi-static setup for 21 days, while mortality, adult growth and reproduction (time of first brood, number of offspring) were observed. The chronic toxicity with regard to mortality, adult growth and reproduction was found to be >= 100 mg/L for each of the aforementioned endpoints. The presence of dissolved material in the different WAF preparations was confirmed prior to the study by GC/MS and NMR analysis as part of a non-GLP approach.

Description of key information

Read across from standolized linseed oil: 21d-NOErLR is 100 mg/l for adult mortality, adult growth and reproduction (time of first brood, number of offspring).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

The chronic (21d-NOELR) toxicity of VEOPOL 212045 (Linseed Standoil 45 poises) towards Daphnia magna with regard to mortality, adult growth and reproduction (time of first brood, number of offspring) is >= 100 mg/L for each of the aforementioned endpoints. This study is read across to this dossier.