Reactionmass of dodecan-2-yl prop-2-enoate and dodecan-3-yl prop-2-enoate and dodecan-4-yl prop-2-enoate

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05th-26th September, 2019

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 3M FMRD, CPDC lot 6
- Expiration date of the lot/batch: 27 Sept, 2021
- Purity: 99.1%:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: In the sample chamber of this facility, stored at room temperature (15-25 C).
- Stability under test conditions: Unstable

Analytical monitoring:

yes

Details on sampling:

- Concentrations: blank control, solvent control, 0.06 mg/L, 0.12 mg/L, 0.25 mg/L, 0.5 mg/L, and 1.0 mg/L
- Sampling method: A sample of ~50 mL was taken from each test solution and control solution. The sample was taken from the leftover test solution for fresh samples and from one replicate for old samples. 10 mL of the sample was extracted before analysis. The remaining sample was stored in a refrigerator for possible re-analysis

Vehicle:

yes

Remarks:

Acetone

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 0.5001 g of the test item was dissolved in 50 mL acetone to get the stock concentration of 1000 mg/L. The stock solution was diluted with acetone to get solvent solutions of 0.6, 1.2, 2.5, and 5.0 mg/m/L. 0.10 mL of solution was added to 1 L of test water to get 0.06, 0.12, 0.25, 0.5 and 1.0 mg/L concentrations. The solutions were stirred with magnetic stirrer (200 rpm) for 1 hour. A solvent control solution was also prepared by adding 0.10 mL acetone into 1L test water and stirred for 1 hour (200 rpm). Tests were preformed with daily renewal of all concentrations.
- Controls: Blank control and solvent control
- Concentration of vehicle in test medium: 100 uL/L

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain/clone: Daphnia magna (Straus)
- Source: Applied Chemistry Department Pesticide Environmental Evaluation Lab of the China Agricultural University
- Age of parental stock: <24 hours old
- Culture conditions: Adult daphnids were cultured in tap water treated with Aquapro Pure Water Systems. During the period preceding the test, water temperatures were 18 - 22°C, photoperiod of 16h light, 8h dark. Daphnids in the cultures were fed daily a suspension of concentrated alga suspension. Neonates were not the first brood.
- Health: The test Daphnia magna was derived from a healthy stock (i.e. showing no signs of stress such as high mortality, presence of males and ephippia, delay in the production of the first brood,
discolored animals, etc.)
- Food type: Living algal cells of Chlorella sp and Desmodesmus subspicatus
- Amount: 0.1 ~ 0.2 mg C/Daphnia/day
- Frequency: Once daily

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

190-200 mg/L CaCO3

Test temperature:

18.9-21.3 C

pH:

8.09-8.42

Dissolved oxygen:

7.25-8.49

Nominal and measured concentrations:

Nominal concentrations: 0.06, 0.12, 0.25, 0.5 and 1.0 mg/L
Measured concentrations: 0.012, 0.017, 0.018, 0.046, 0.061 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 200 or 250-mL glass beakers
- Type: closed
- Fill volume: 100 mL per vessel
- Aeration: Water was aerated for more than 24 hours before use to attain dissolved oxygen saturation of at least 80% ASV
-Renewal rate: 24 hours
- No. of organisms per vessel: One
- No. of vessels per concentration: Ten
- No. of vessels per control: Ten
- No. of vessels per vehicle control: Ten

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water treated with Aquapro Pure Water System
- Total organic carbon:<2 mg/L, measured every three months.
- Alkalinity: 140 - 250 mg CaCO3/L
- Particulate matter: <5 mg/L suspended solids
- Metals:All less than detection limits
- Pesticides:all less than detection limits
- Culture medium different from test medium: No

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16h light, 8h dark
- Light intensity: 1000 ~ 1500 lux for cool white light


EFFECT PARAMETERS MEASURED: Effects measured during the definitive test include total number of broods per parent, mortality of parents daphnids, body length of parent daphnids, time to first brood, and total living offspring daphnid per parent. These parameters were measured daily. In addition, number of aborted eggs or dead offspring were recorded daily.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2-fold
- Range finding study: yes
- Test concentrations: Nominal concentrations: 1.0, 3.0 and 10 mg/L
- Results used to determine the conditions for the definitive study: In the 10 mg/L test group, all daphnids were dead after 24 hours. Actual concentration at this level was about 3 mg/L.The daphnids in other concentrations were normal after 6 days. The maximum concentration for the definitive test was 1 mg/L based on this result.

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.06 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.012 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

- Reproduction data summary in Table 2
-Time until first brood summary in Table 3, NOEC was 0.06 mg/L (nominal)
-Body Length summary in Table 4
-Broods per parent summary in Table 5
- NOEC for reproduction, parent body length, time until first brood, and total brood per parents, 0.06 mg/L (nominal), 0.012 mg/L (measured)
- NOEC for parent mortality, 0.25 mg/L (nominal), 0.018 mg/L (measured)
- Behaviorall abnormalities: None
- Other: During the test, there were no dead offspring, aborted eggs, or male offspring.
- Mortality of control: No mortality in control Daphnids (0%), within guideline criteria
- Effect concentrations exceeding solubility of substance in test medium: No

Reported statistics and error estimates:

Statistical analyses were performed on survival of first-generation daphnids, the number of live young and the length and dry weight of the surviving first-generation daphnids. The F-test was used to analyze homogeneity of variance for live offspring, first brood time, total # of broods, and parental body length. Treatment groups with significant survival effects were excluded from the analyses of reproduction and growth. Reproduction and growth (length) data were evaluated to determine significance via a one-side Student´s T-test using a 5% significance level in an EXCEL document. If the P-value in the F-test was <0.05, and unequal-variance T-test was done.

Table 2. Total Juveniles produced per Parent during Test.

Nominal concentrations	Mean ± SD	P- value of F test	P-value of T-Test	CV (%)	Inhibition Rate   (%)
Control	266 ± 26	-	-	9.8%	-
Solvent control	263 ± 28	0.79	0.86	10.6%	1.1%
0.06	248 ± 22	0.47	0.095	8.9%	6.8%
0.12	147 ± 22***	0.50	3.2E-09	15.0%	44.7%
0.25	35 ± 13***	0.03	5.6E-12 ¹	37.1%	86.8%
0.5	7 ± 8***	0.001	3.8E-12 ¹	114.3%	97.4%
1.0	-	-	-	-	-

“***”P<0.001

¹Unequal-variance T-test due non-homogeneity of variance in F-test (P<0.05).

 

Table 3. First Brood Time of Each Parent (day)

Nominal concentrations	Mean ± SD (days)	P- value of F test	P-value of T-Test
Control	7.6 ± 0.5	-	-
Solvent control	7.7 ± 0.5	0.85	0.66
0.06	7.9 ± 0.3	0.22	0.07
0.12	9.2 ± 0.6***	0.43	3.8E-06
0.25	9.3 ± 1.6***	0.001 ¹	0.005
0.5	10.3 ± 2.9*	0.00002 ¹	0.034
1.0	N/A	N/A	N/A

Notes: First brood time means day first brood was observed; N/A means not applicable.

“*” p<0.05. “**” p<0.01, “***” p<0.001

¹Unequal-variance T-test due non-homogeneity of variance in F-test (P<0.05).

 

Table 4. Body Length (mm) of Parent

Nominal concentrations	Mean ± SD	P- value of F test	P-value of T-Test
Control	3.44 ± 0.14	-	-
Solvent control	3.35 ± 0.12	0.59	0.14
0.06	3.29 ± 0.08	0.27	0.12
0.12	2.86 ± 0.14***	0.60	6.4E-08
0.25	2.15 ± 0.10***	0.59	1.4E-15
0.5	1.96 ± 0.10***	0.99	7.8E-10
1.0	N/A	N/A	N/A

N/A means not applicable.

“***” P<0.001

 

Table 5. Number of Broods per Parent

Nominal concentrations	Mean ± SD	P- value of F test	P-value of T-Test
Control	6.3 ± 0.5	-	-
Solvent control	6.0 ± 0.0	N/A	N/A
0.06	6.1 ± 0.3	0.22	0.14
0.12	5.3 ± 0.5***	1.0	1.0E-04
0.25	3.8 ± 1.1***	0.02	1.6E-05 ¹
0.5	1.0 ± 1.1***	0.03	1.6E-09 ¹
1.0	N/A	N/A	N/A

N/A means not applicable.

“***” p<0.001

1, Unequal-variance T-test due non-homogeneity of variance in F-test (P<0.05).

 

Validity criteria fulfilled:

yes

Remarks:

Mortality of the parent animals in the controls did not exceed 20% at the end of the test and the mean number of living offspring from the parents in the controls was ≥ 60.

Conclusions:

21 day NOEC = 0.06 mg/L (OECD 211) in Daphnia magna (nominal) for reproduction.
21 day NOEC = 0.012 mg/L (OECD 211) in Daphnia magna (measured) for reproduction.

Executive summary:

The 21 day NOEC to Daphnia magna was determined in a semi-static definitive test according to OECD 211 guidelines. Nominal concentrations of 0.06, 0.12, 0.25, 0.5 and 1 mg/L, a solvent control, and a blank control were run using 1 animals per vessel (10 replicates per concentration) and acetone as a dispersant at 100 ug/L. Measured geometric mean concentrations of the substance during the test were : 0.012, 0.017, 0.018, 0.046, 0.061 mg/L. A NOEC of 0.06 mg/L (nominal concentration) and NOEC of 0.012 mg/L (measured) was determined.

Study was conducted under international guidelines and was compliant with GLP criteria. Test substance concentrations were not maintained at >=80% of the nominal throughout the test. However, the test was conducted with a semi static method and is therefore considered reliable with restrictions. The result is considered suitable for use in Risk Assessment, Classification & Labeling, and PBT analysis.

Description of key information

21 day NOEC = 0.06 mg/L (OECD 211) in Daphnia magna (nominal) for reproduction.

21 day NOEC = 0.012 mg/L (OECD 211) in Daphnia magna (measured) for reproduction.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.06 mg/L

Additional information

The 21 day NOEC to Daphnia magna was determined in a semi-static definitive test according to OECD 211 guidelines. Nominal concentrations of 0.06, 0.12, 0.25, 0.5 and 1 mg/L, a solvent control, and a blank control were run using 1 animals per vessel (10 replicates per concentration) and acetone as a dispersant at 100 ug/L. Measured geometric mean concentrations of the substance during the test were : 0.012, 0.017, 0.018, 0.046, 0.061 mg/L. A NOEC of 0.06 mg/L (nominal concentration) and NOEC of 0.012 mg/L (measured) was determined.

Study was conducted under international guidelines and was compliant with GLP criteria. Test substance concentrations were not maintained at >=80% of the nominal throughout the test. However, the test was conducted with a semi static method and is therefore considered reliable with restrictions. The result is considered suitable for use in Risk Assessment, Classification & Labeling, and PBT analysis.