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EC number: 256-458-9 | CAS number: 49744-28-7
- 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
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2017-11-17 to 2017-12-07
- Reliability:
- 1 (reliable without restriction)
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Deviations:
- no
- GLP compliance:
- yes
Test material
- Reference substance name:
- 1-[(4-methoxy-2-nitrophenyl)azo]-2-naphthol
- EC Number:
- 256-458-9
- EC Name:
- 1-[(4-methoxy-2-nitrophenyl)azo]-2-naphthol
- Cas Number:
- 49744-28-7
- Molecular formula:
- C17H13N3O4
- IUPAC Name:
- 1-[(4-methoxy-2-nitrophenyl)diazenyl]-2-naphthol
- Test material form:
- solid: particulate/powder
- Details on test material:
- Batch number
3. April 2010
CAS number
49744-28-7
Sum formula
C17H13N3O4
Molecular weight
323.31 g/mol
Purity of main component (certified)
98.50% (w/w) (comp. 1)
Water solubility
3.3 µg/L
Appearance
Red powder
pH value
9.1 (10% H2O)
Stability
Stable in water and light at room temperature
Expiry date
2020-04-03
Recommended storage
Room temperature
Constituent 1
Sampling and analysis
- Analytical monitoring:
- no
- Remarks:
- No analytical monitoring was carried out due to the low solubility of the test item .
Test solutions
- Vehicle:
- no
- Details on test solutions:
- Preparation of the saturated solution
The saturated solution (1 mg/L test item was weighed out) was prepared with demineralized water three days prior to the start of the exposure (0 h). The dispersion was stirred for 72 hours (1100 rpm, room temperature) with a magnetic stirrer. Undissolved particles were removed by membrane filtration (membrane filter 0.45 µm, RC, MACHEREY-NAGEL). The filter was saturated in order to avoid adsorption during the filtration. The first 25 mL filtrate were discarded. The filtration was interrupted for ca. 15 minutes to allow for adsorption and saturation of the filter material with dissolved test item. Thereafter, the filtration was continued. The next 25 mL were discarded. After the separation treatment the components of the dilution water were added to the saturated solution. The following filtrate, i.e. the saturated solution, was used in the test. During filtration the filter was always kept covered.
Test concentrations (nominal)
Five dilution levels out of the saturated solution were tested in a geometrical series with a dilution factor of 2.50: 2.56 - 6.40 - 16.0 - 40.0 - 100% of the saturated solution. The dilution levels are based on the results of a preliminary range finding test (non-GLP, Erlenmeyer flasks, with headspace in OECD TG 201 medium)
Control
Six replicates (without test item) were tested under the same test conditions as the test item replicates.
Test organisms
- 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 corresponding to 35 - 70 µE ∙ m-2 ∙ s-1 for 24 h per day.
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Post exposure observation period:
- No
Test conditions
- Hardness:
- Not specified
- Test temperature:
- min.: 22.0 max.: 23.0 mean value: 22.5
- pH:
- Nominal test item concentration pH-value
[% saturated solution] Start; 0 hours End; 72 hours
100 7.74 9.09
40.0 7.73 9.32
16.0 7.80 9.25
6.40 7.86 9.16
2.56 8.06 9.19
Control 8.17 9.16 - Dissolved oxygen:
- Not measured
- Salinity:
- Not measured, freshwater conditions
- 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 2 - 5 x 10^3 cells/mL, Actual: mean 6839 cells/mL
- Control end cells density: Mean 2253449 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 0.064a2EDTA 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: Approximately 4440 to 8880 lux, corresponding to 60 to 120
µE ∙ m-2 ∙ s-1, mean value: 6132 lux,
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
-Chlorophyll a-fluorescence
The cell density was measured daily via Chlorophyll a-fluorescence, excitation at 436 nm, emission at 685 nm. Dilution water was used as a background signal. No self-fluorescence was found in the preliminary range finding test at the saturated solution. - Reference substance (positive control):
- yes
- Remarks:
- (Potassium dichromate)
Results and discussion
Effect concentrationsopen allclose all
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 other: % saturated solution
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: growth rate und yield
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- > 100 other: % saturated solution
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: growth rate and yield
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 other: % saturated solution
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 other: % saturated solution
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- 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: The test media clear throughout the exposure period.
- 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 2017-10-10 to 2017-10-13. 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 0.535 0.745 ± 0.524
95 % confidence interval 0.523 – 0.547
Yield inhibition
EyC50 0.320 0.398 ± 0.291
95 % confidence interval 0.249 – 0.339
SD = Standard deviation - Reported statistics and error estimates:
- EC-values and statisticall analyses
EC10-, EC20- and EC50-values with confidence intervals of growth rate and yield inhibition after 72 hours were determined from the recorded data without any statistical calculations.
NOEC, LOEC and statistically analyses
The NOEC and LOEC were determined by calculation of significant differences of growth rates and yield. As a standard, One Way Analysis of Variance (ANOVA) was used for NOEC/LOEC calculations. When running a One Way Analysis of Variance, a Normality test and an Equal Variance test were done first. P-values for both Normality and Equal Variance tests are 0.05. The -value (acceptable probability of incorrectly concluding that there is a difference) is =0.05. The Normality test failed for calculation of growth rate. Therefore, the data were transformed (Y=10Y).
Software
The data for the tables in this report were computer-generated and have been rounded for presentation from the full 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.
- GraphPad Prism, GRAPHPAD SOFTWARE, INC.
- ToxRat Professional, Version 3.2, TOXRAT SOLUTIONS GMBH
Any other information on results incl. tables
Cell Densities
Nominal test item concentration
[% saturated solution] |
Replicate |
Cell density [cells/mL] |
|||||||||
No. |
0 hours |
24 hours |
48 hours |
72 hours |
|||||||
100 |
1 |
6839 |
28927 |
339712 |
2287761 |
||||||
2 |
6839 |
28389 |
297449 |
2252298 |
|||||||
3 |
6839 |
34182 |
329694 |
2450740 |
|||||||
Mean |
6839 |
30499 |
322285 |
2330266 |
|||||||
40.0 |
1 |
6839 |
33293 |
353078 |
2529683 |
||||||
2 |
6839 |
35394 |
402668 |
2771546 |
|||||||
3 |
6839 |
32834 |
390145 |
2609095 |
|||||||
Mean |
6839 |
33840 |
381964 |
2636775 |
|||||||
16.0 |
1 |
6839 |
35160 |
421687 |
2508733 |
||||||
2 |
6839 |
32918 |
389911 |
2402812 |
|||||||
3 |
6839 |
33217 |
435369 |
2696933 |
|||||||
Mean |
6839 |
33765 |
415656 |
2536159 |
|||||||
6.40 |
1 |
6839 |
28117 |
344827 |
1819367 |
||||||
2 |
6839 |
31455 |
359914 |
2545015 |
|||||||
3 |
6839 |
29185 |
348385 |
2197933 |
|||||||
Mean |
6839 |
29586 |
351042 |
2187438 |
|||||||
2.56 |
1 |
6839 |
34235 |
355782 |
2271083 |
||||||
2 |
6839 |
29750 |
344488 |
2284660 |
|||||||
3 |
6839 |
27751 |
341304 |
2256336 |
|||||||
Mean |
6839 |
30579 |
347191 |
2270693 |
|||||||
Control |
1 |
6839 |
28881 |
252752 |
1931550 |
||||||
2 |
6839 |
39935 |
459901 |
2485266 |
|||||||
3 |
6839 |
33302 |
373923 |
1288708 |
|||||||
4 |
6839 |
31657 |
388705 |
1934710 |
|||||||
5 |
6839 |
33226 |
457349 |
2978824 |
|||||||
6 |
6839 |
33846 |
385276 |
2901636 |
|||||||
Mean |
6839 |
33475 |
386318 |
2253449 |
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 item concentration |
Replicate |
Growth rate |
Inhibition of growth rate |
Yield |
Inhibition of yield |
||||||||||
[% saturated solution] |
No. |
[d-1] |
[%] |
[cells/mL] |
[%] |
||||||||||
100 |
1 |
|
1.94 |
-1 |
|
2280922 |
-2 |
||||||||
2 |
|
1.93 |
-1 |
|
2245459 |
0 |
|||||||||
3 |
|
1.96 |
-2 |
|
2443901 |
-9 |
|||||||||
Mean |
(-) |
1.94 |
-1 |
(-) |
2323427 |
-3 |
|||||||||
40.0 |
1 |
|
1.97 |
-3 |
|
2522844 |
-12 |
||||||||
2 |
|
2.00 |
-4 |
|
2764707 |
-23 |
|||||||||
3 |
|
1.98 |
-3 |
|
2602256 |
-16 |
|||||||||
Mean |
(-) |
1.99 |
-3 |
(-) |
2629936 |
-17 |
|||||||||
16.0 |
1 |
|
1.97 |
-3 |
|
2501894 |
-11 |
||||||||
2 |
|
1.95 |
-2 |
|
2395973 |
-7 |
|||||||||
3 |
|
1.99 |
-4 |
|
2690094 |
-20 |
|||||||||
Mean |
(-) |
1.97 |
-3 |
(-) |
2529320 |
-13 |
|||||||||
6.40 |
1 |
|
1.86 |
3 |
|
1812528 |
19 |
||||||||
2 |
|
1.97 |
-3 |
|
2538176 |
-13 |
|||||||||
3 |
|
1.92 |
0 |
|
2191094 |
2 |
|||||||||
Mean |
(-) |
1.92 |
0 |
(-) |
2180599 |
3 |
|||||||||
2.56 |
1 |
|
1.94 |
-1 |
|
2264244 |
-1 |
||||||||
2 |
|
1.94 |
-1 |
|
2277821 |
-1 |
|||||||||
3 |
|
1.93 |
-1 |
|
2249497 |
0 |
|||||||||
Mean |
(-) |
1.94 |
-1 |
(-) |
2263854 |
-1 |
|||||||||
Control |
1 |
|
1.88 |
|
|
1924711 |
|
||||||||
2 |
|
1.97 |
|
|
2478427 |
|
|||||||||
3 |
|
1.75 |
|
|
1281869 |
|
|||||||||
4 |
|
1.88 |
|
|
1927871 |
|
|||||||||
5 |
|
2.03 |
|
|
2971985 |
|
|||||||||
6 |
|
2.02 |
|
|
2894797 |
|
|||||||||
Mean |
|
1.92 |
|
|
2246610 |
|
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 |
1.44 |
2.17 |
2.03 |
1.88 |
0.388 |
20.6 |
25.4 |
2 |
1.77 |
2.44 |
1.69 |
1.97 |
0.416 |
21.2 |
||
3 |
1.58 |
2.42 |
1.24 |
1.75 |
0.607 |
34.8 |
||
4 |
1.53 |
2.51 |
1.61 |
1.88 |
0.544 |
28.9 |
||
5 |
1.58 |
2.62 |
1.87 |
2.03 |
0.537 |
26.5 |
||
6 |
1.60 |
2.43 |
2.02 |
2.02 |
0.417 |
20.7 |
||
|
|
|
Mean |
1.92 |
|
|
||
|
|
|
SD ± |
0.11 |
|
|||
|
|
|
CV [%] |
5.50 |
|
SD = Standard deviation CV = Coefficient of variation
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Conclusions:
- In this study, 1-((4-Methoxy-2-nitrophenyl)azo)-2-naphthol was found not to inhibit the growth of the freshwater green alga Pseudokirchneriella subcapitata after 72 hours with the following effect values (based on nominal test item concentrations): The EC50-values for both inhibition of growth rate (ErC50) and yield (EyC50) after 72 hours were > 100 % saturated solution, respectively. The NOEC-values for both inhibition of growth rate and yield after 72 hours were 100 % saturated solution, respectively. The saturated solution was prepared with a nominal loading of 1 mg/L.
- Executive summary:
The toxicity of 1-((4-Methoxy-2-nitrophenyl)azo)-2-naphthol (batch number:3.April 2010 g) to the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined according to the principles of OECD 201 and Council Regulation (EC) No. 266/2016/Method C.3 from 2017-11-17 to 2017-12-07 at the test facility. The aim of the study was to assess the effects on growth rate and yield over a period of 72 hours.
The study was conducted under static conditions with an initial cell density of 6839 cells/mL. A saturated solution with a nominal loading of 1.00 mg test item/L was tested. Five concentrations were tested in a geometrical series with a dilution factor of 2.50 (nominal):2.56 - 6.40 - 16.0 - 40.0 - 100 % saturated solution. Three replicates were tested for each test item concentration and six replicates for the control. The environmental conditions were within the acceptable limits.
No analytical monitoring was carried out due to the low solubility of the test item.
All effect values were based on the nominal test item of1-((4-Methoxy-2-nitrophenyl)azo)-2-naphthol. The results are summarized in the following table:
NOEC, LOEC, EC - values of 1-((4-Methoxy-2-nitrophenyl)azo)-2-naphthol (0-72hours)
based on the nominal test item concentration [% saturated solution]
Nominal test item concentration
[% saturated solution]Inhibition of Growth Rate
NOEC
100
LOEC
> 100
ErC10
> 100
ErC20
> 100
ErC50
> 100
Inhibition of Yield
NOEC
100
LOEC
> 100
EyC10
> 100
EyC20
> 100
EyC50
> 100
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