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EC number: 612-975-5 | CAS number: 6225-08-7
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Toxicity to aquatic algae and cyanobacteria
Administrative data
Link to relevant study record(s)
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2016-12-12 to 2016-12-15
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Deviations:
- no
- GLP compliance:
- yes
- Analytical monitoring:
- yes
- Details on sampling:
- Determination of the test item
All concentration levels and the control were analytically verified via LC-MS/MS at the start (0 hours) and at the end of the exposure (72 hours) with algae. - Vehicle:
- no
- Details on test solutions:
- Stock solution
100 mg test item/L were freshly prepared with dilution water. The dispersion was agitated until the stock solution was clear.
Test concentrations
5 concentrations were tested in a geometrical series with a dilution factor of √10: 1.00 - 3.16 - 10.0 - 31.6 - 100 mg test item/L. The concentrations are based on the results of a preliminary range finding test (non-GLP). - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata HINDÁK
- Strain: SAG 61.81
- Source (laboratory, culture collection): SAG, Pflanzenphysiologisches Institut der Universität Göttingen, Nikolausberger Weg 18, D-37073 Göttingen
- Age of inoculum (at test initiation): 3 days
- Method of cultivation: Fresh stocks were prepared every month on Z-Agar. Light intensity amounted to 2567 – 5130 lux for 24 h per day. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Post exposure observation period:
- No
- Hardness:
- Not specified
- Test temperature:
- min.: 22.0 max.: 23.0 mean value: 22.5
- pH:
- Nominal test item concentration pH-value
[mg/L] Start; 0 hours End; 72 hours
100 7.46 8.12
31.6 7.51 8.32
10.0 7.58 9.49
3.16 7.68 9.64
1.00 7.79 9.51
Control 7.97 9.42 - Dissolved oxygen:
- Not measured
- Salinity:
- Not measured, freshwater conditions
- Nominal and measured concentrations:
- Nominal 1.00 - 3.16 - 10.0 - 31.6 - 100 mg/L
- Details on test conditions:
- TEST SYSTEM
- Test vessel: 250 mL Erlenmeyer flasks
- Type (delete if not applicable): open, cotton wool plugs
- Material, size, headspace, fill volume: sterile 250 mL Erlenmeyer flasks, test volume 100 mL
- Aeration: Test containers were placed on a rotary shaker and oscillated at appr. 70 rpm
- Type of flow-through (e.g. peristaltic or proportional diluter): None
- Renewal rate of test solution (frequency/flow rate): One application at test start
- Initial cells density: Nominal: approximately 5 x 10^3 - 10^4 cells/mL, Actual: mean 6305 cells/mL
- Control end cells density: Mean 2330531 cells/mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
GROWTH MEDIUM
- Standard medium used: yes
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Composition of Dilution Water
according to the guideline
Component Concentration [mg/L]
NH4Cl 15
MgCl2 x 6 H2O 12
CaCl2 x 2 H2O 18
MgSO4 x 7 H2O 15
KH2PO4 1.6
FeCl3 x 6 H2O 0.064
Na2EDTA x 2 H2O 0.1
H3BO3 0.185
MnCl2 x 4 H2O 0.415
ZnCl2 3 x 10^-3
Na2MoO4 x 2 H2O 7 x 10^-3
CoCl2 x 6 H2O 1.5 x 10^-3
CuCl2 x 2 H2O 1 x 10^-5
NaHCO3 50
pH 8.1 ± 0.2
OTHER TEST CONDITIONS
- Sterile test conditions: no
- Adjustment of pH: no
- Photoperiod: 24 h
- Light intensity and quality: 4440 - 8880 lux, mean value: 5035 lux,
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: [counting chamber; electronic particle counter; fluorimeter; spectrophotometer; colorimeter] fluorimeter, Cell density and self fluorescence was measured daily via Chlorophyll-a- fluorescence, excitation at 436 nm, emission at 685 nm. Dilution water was used as background signal. - Reference substance (positive control):
- yes
- Remarks:
- (Potassium dichromate)
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 3.16 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 16.3 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: CI: 14.6 - 18.2
- Duration:
- 72 h
- Dose descriptor:
- EC20
- Effect conc.:
- 20.1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: CI: 18.5 - 21.6
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 28.8 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: CI: 28.2 - 29.3
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 3.16 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Yield
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 10.2 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Remarks on result:
- other: CI: 9.29 - 11.7
- Duration:
- 72 h
- Dose descriptor:
- EC20
- Effect conc.:
- 11.9 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Remarks on result:
- other: CI: 10.9 - 14.3
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 15.6 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Yield
- Remarks on result:
- other: CI: 13.7 - 19.2
- Details on results:
- - Exponential growth in the control (for algal test): yes
- Observation of abnormalities (for algal test):
- Unusual cell shape: No
- Colour differences: No
- Flocculation: No
- Adherence to test vessels: No
- Aggregation of algal cells: No
- Other: No
- Effect concentrations exceeding solubility of substance in test medium: No - Results with reference substance (positive control):
- The toxicity of potassium dichromate (SIGMA ALDRICH, batch number MKBV0900V, purity 99.0 %, CAS RN 7778-50-9) to the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined over a period of 72 hours from 2016-10-11 to 2016-10-14 (with headspace). The reference item toxicity is in the valid range following test facility SOPS.
EC50-Values of the Reference Item
based on nominal concentrations mg/L, (0-72 hours)
Current Study Valid Range (average ± 3 x SD)
Growth Rate inhibition
ErC50 0.435 0.763 ± 0.514
95% confidence interval 0.414 – 0.458
Yield inhibition
EyC50 0.230 0.402 ± 0.293
95% confidence interval 0.179 – 0.322
SD = Standard deviation - Reported statistics and error estimates:
- EC-values and statistical analyses
EC10-, EC20- and EC,50-values with confidence intervals of growth rate and yield inhibition after 72 hours were calculated by sigmoidal dose-response regression.
NOEC, LOEC and statistical analyses
The NOEC and LOEC were determined by calculation of statistically significant differences of growth rates and yield.
The following statistical tests were conducted:
Monotonicity was tested by trend analysis by contrasts (significance level 0.05).
Shapiro-Wilk’s test on normal distribution was done with a significance level to 0.01.
Levene’s test on variance homogeneity was done with a significance level to 0.01.
William’s test on significant differences was done for growth rate with a significance level to 0.05.
Multiple sequentially-rejective welsh-t-test after Bonferroni-holm was done for yield with a significance level to 0.05.
Software
The data for the tables in this report were computer-generated and have been rounded for presentation from the fully derived data. Consequently, if calculated manually based on the given data minor deviations may occur from these figures.
Calculations were carried out using software
• Excel, MICROSOFT CORPORATION
•SigmaPlot, SPSS INC.
•ToxRat, TOXRAT SOLUTIONS GMBH
•GraphPad Prism, GRAPHPAD SOFTWARE, INC. - Validity criteria fulfilled:
- yes
- Conclusions:
- In this study, N,N-Dimethylnonanamide was found to inhibit the growth of the freshwater green alga Pseudokirchneriella subcapitata after 72 hours with the following effect values (based on the nominal test item concentrations): The NOEC (0-72 h) for inhibition of growth and inhibition of yield was 3.16 mg/L. The EC50-values with 95 % confidence intervals for inhibition of growth rate (ErC50) and yield (EyC50) after 72 hours were 28.8 (28.2 – 29.3) mg/L and 15.6 (13.7 – 19.2) mg/L, respectively. All effect values are based on the nominal test item concentrations of N,N-Dimethylnonanamide.
- Executive summary:
The toxicity of N,N-Dimethylnonanamide to the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined according to the principles of OECD 201 andCouncil Regulation (EC) No. 761/2009 Method C.3 from2016-12-12 to 2016-12-15 at the test facility. The aim of the study was the determination of NOEC, LOEC, EC10- , EC20- and EC50-values of growth rate and yield over a period of 72 hours.
The study was conducted under static conditions with an initial cell density of 6305 cells/mL. A stock solution of 100 mg/L was freshly prepared with dilution water. Five concentrations were tested in a geometrical series with a dilution factor of √10:1.00 - 3.16 - 10.0 - 31.6 - 100 mg/L.The environmental conditions were within the acceptable limits.
The test media were visually clear throughout the test period. The concentrations of thetest item N,N-Dimethylnonanamide in all test concentrations and the control were analytically verified by LC-MS/MS at the start and the end of exposure.
The measured concentrations of N,N-Dimethylnonanamidein the fresh media (0 h) were in the range of 98 to 109% of the nominal values at all tested concentration levels. The measured concentrations in the old media (72 h) were in the range of 88 to 105% of the nominal values of all concentrations.Therefore, all effect values given are based on the nominal test item concentrations of N,N-Dimethylnonanamide.
NOEC, LOEC, EC-values and 95% Confidence Intervals ofN,N-Dimethylnonanamide(0 - 72 hours)
based on nominal test item concentrations [mg/L]
Inhibition of Growth Rate
NOEC
3.16
LOEC
10.0
ErC10
16.3 (14.6 – 18.2)
ErC20
20.1 (18.5 – 21.6)
ErC50
28.8 (28.2 – 29.3)
Inhibition of Yield
NOEC
3.16
LOEC
10.0
EyC10
10.2 (9.29 – 11.7)
EyC20
11.9 (10.9 – 14.3)
EyC50
15.6 (13.7 – 19.2)
Reference
Cell Densities
Nominal test itemconcentration |
Replicate |
Cell density [cells/mL] |
|||
[mg/L] |
No. |
0 hours |
24 hours |
48 hours |
72 hours |
100 |
1 |
6305 |
< LOQcell density |
< LOQcell density |
< LOQcell density |
2 |
6305 |
< LOQcell density |
< LOQcell density |
< LOQcell density |
|
3 |
6305 |
< LOQcell density |
< LOQcell density |
< LOQcell density |
|
Mean |
6305 |
n.a. |
n.a. |
n.a. |
|
31.6 |
1 |
6305 |
12250 |
24293 |
71745 |
2 |
6305 |
10522 |
18618 |
65761 |
|
3 |
6305 |
14525 |
25821 |
78618 |
|
Mean |
6305 |
12432 |
22911 |
72041 |
|
10.0 |
1 |
6305 |
35222 |
324638 |
2124603 |
2 |
6305 |
45092 |
320804 |
2158769 |
|
3 |
6305 |
38064 |
385325 |
2090324 |
|
Mean |
6305 |
39459 |
343589 |
2124565 |
|
3.16 |
1 |
6305 |
46901 |
430531 |
2238829 |
2 |
6305 |
60808 |
549538 |
2364444 |
|
3 |
6305 |
42662 |
447625 |
2396467 |
|
Mean |
6305 |
50124 |
475898 |
2333247 |
|
1.00 |
1 |
6305 |
48037 |
493439 |
2498966 |
2 |
6305 |
54200 |
449542 |
2435257 |
|
3 |
6305 |
42174 |
458698 |
2183295 |
|
Mean |
6305 |
48137 |
467226 |
2372506 |
|
Control |
1 |
6305 |
38960 |
395541 |
2381527 |
2 |
6305 |
42774 |
342589 |
2266850 |
|
3 |
6305 |
46431 |
451989 |
2452340 |
|
4 |
6305 |
45551 |
399251 |
2296731 |
|
5 |
6305 |
50862 |
413741 |
2331010 |
|
6 |
6305 |
51730 |
484982 |
2292559 |
|
Mean |
6305 |
46051 |
414682 |
2336836 |
< LOQcell density= below the limit of quantification (2736 cells/mL) n.a. = data not determinable
Evaluation after 72 hours
Statistically
significant differences of growth rates and yield compared to
control
values are marked (+), not significant differences are marked (-).
Nominal test itemconcentration |
Replicate |
Growth rate |
Inhibition of growth rate |
Yield |
Inhibition of yield |
||
[mg/L] |
No. |
[d-1] |
[%] |
[cells/mL] |
[%] |
||
100 |
1 |
|
n.a. |
100 |
|
n.a. |
100 |
2 |
|
n.a. |
100 |
|
n.a. |
100 |
|
3 |
|
n.a. |
100 |
|
n.a. |
100 |
|
Mean |
(+) |
n.a. |
100 |
(+) |
n.a. |
100 |
|
31.6 |
1 |
|
0.811 |
59 |
|
65440 |
97 |
2 |
|
0.782 |
60 |
|
59456 |
97 |
|
3 |
|
0.841 |
57 |
|
72313 |
97 |
|
Mean |
(+) |
0.811 |
59 |
(+) |
65736 |
97 |
|
10.0 |
1 |
|
1.94 |
2 |
|
2118298 |
9 |
2 |
|
1.95 |
1 |
|
2152464 |
8 |
|
3 |
|
1.94 |
2 |
|
2084019 |
11 |
|
Mean |
(+) |
1.94 |
2 |
(+) |
2118260 |
9 |
|
3.16 |
1 |
|
1.96 |
1 |
|
2232524 |
4 |
2 |
|
1.98 |
0 |
|
2358139 |
-1 |
|
3 |
|
1.98 |
0 |
|
2390162 |
-3 |
|
Mean |
(-) |
1.97 |
0 |
(-) |
2326942 |
0 |
|
1.00 |
1 |
|
1.99 |
-1 |
|
2492661 |
-7 |
2 |
|
1.99 |
-1 |
|
2428952 |
-4 |
|
3 |
|
1.95 |
1 |
|
2176990 |
7 |
|
Mean |
(-) |
1.98 |
0 |
(-) |
2366201 |
-2 |
|
Control |
1 |
|
1.98 |
|
|
2375222 |
|
2 |
|
1.96 |
|
|
2260545 |
|
|
3 |
|
1.99 |
|
|
2446035 |
|
|
4 |
|
1.97 |
|
|
2290426 |
|
|
5 |
|
1.97 |
|
|
2324705 |
|
|
6 |
|
1.97 |
|
|
2286254 |
|
|
Mean |
|
1.97 |
|
|
2330531 |
|
negative inhibition = increase of growth
n.a. = not available
Section-by-Section and Average Specific Growth Ratesof the Control Group (0 – 72 hours)
|
Replicate No. |
Specific Growth Rate [d-1] |
Mean (0 - 72 hours) |
SD ± |
CV |
Mean CV [%] |
||
section-by-section |
||||||||
0 - 24 hours |
24 - 48 hours |
48 - 72 hours |
||||||
Control |
1 |
1.82 |
2.32 |
1.80 |
1.98 |
0.294 |
14.9 |
12.5 |
2 |
1.92 |
2.08 |
1.89 |
1.96 |
0.104 |
5.3 |
||
3 |
2.00 |
2.28 |
1.69 |
1.99 |
0.292 |
14.7 |
||
4 |
1.98 |
2.17 |
1.75 |
1.97 |
0.211 |
10.7 |
||
5 |
2.09 |
2.10 |
1.73 |
1.97 |
0.210 |
10.6 |
||
6 |
2.11 |
2.24 |
1.55 |
1.97 |
0.363 |
18.5 |
||
|
|
|
Mean |
1.97 |
|
|
||
|
|
|
SD ± |
0.01 |
|
|||
|
|
|
CV [%] |
0.49 |
|
SD = Standard deviation CV = Coefficient of variation
Description of key information
In this study, N,N-Dimethylnonanamide was found to inhibit the growth of the freshwater green alga Pseudokirchneriella subcapitata after 72 hours with the following effect values (based on the nominal test item concentrations): The NOEC (0-72 h) for inhibition of growth and inhibition of yield was 3.16 mg/L. The EC50-values with 95 % confidence intervals for inhibition of growth rate (ErC50) and yield (EyC50) after 72 hours were 28.8 (28.2 – 29.3) mg/L and 15.6 (13.7 – 19.2) mg/L, respectively. All effect values are based on the nominal test item concentrations of N,N-Dimethylnonanamide.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 28.8 mg/L
- EC10 or NOEC for freshwater algae:
- 16.3 mg/L
Additional information
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Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.