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EC number: 215-520-5 | CAS number: 1328-25-2 This substance is identified in the Colour Index by Colour Index Constitution Number, C.I. 65230.
- 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 plants other than algae
Administrative data
- Endpoint:
- toxicity to aquatic plants other than algae
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From 2017-10-05 to 2017-10-17, with the definitive exposure phase from 2017-10-06 to 2017-10-13
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 017
- Report date:
- 2017
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 221 (Lemna sp. Growth Inhibition Test)
- Version / remarks:
- 2006
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Anthra[9,1,2-cde]benzo[rst]pentaphene-5,10-dione, amino-, reaction products with 1-amino-9,10-anthracenedione and tetrabromo-8,16-pyranthrenedione
- EC Number:
- 215-520-5
- EC Name:
- Anthra[9,1,2-cde]benzo[rst]pentaphene-5,10-dione, amino-, reaction products with 1-amino-9,10-anthracenedione and tetrabromo-8,16-pyranthrenedione
- Cas Number:
- 1328-25-2
- Molecular formula:
- C126H58N4O10
- IUPAC Name:
- anthra[9,1,2-cde]benzo[rst]pentaphene-5,10-dione, amino-, reaction products with 1-amino-9,10-anthracenedione and tetrabromo-8,16-pyranthrenedione
- Test material form:
- solid: particulate/powder
Constituent 1
Sampling and analysis
- Analytical monitoring:
- no
- Details on sampling:
- No analytical monitoring was carried out due to the low water solubility of the test item.
Test solutions
- Vehicle:
- no
- Details on test solutions:
- Preparation of the Test item solution
A test item solution of 100 mg test item/L was prepared once 24 ± 1 hour prior to the start of the exposure. An appropriate amount of the test item was weighed out. The test item was applied onto a glass slide. The glass slide with the test item was inserted into a glass bottle with an appropriate amount of dilution water. The test item solution was stirred for 24 ± 1 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 of the filtrate were discarded. The filtration was interrupted for 10 minutes to allow adsorption and saturation of the filter material with dissolved test item. Thereafter, the filtration was continued. The next 25 mL were discarded. The following filtrate, i.e. the test item solution, was used as a test item solution in the test. During filtration, the filter was always kept covered. The test item solution was checked via laser beam (Tyndall effect) for undissolved test item, which was negative. The concentrations are based on the results of a preliminary range finding test (see Annex I).
Test loading
All terms related to concentration level was given as loading level because partly dissolved compounds and mixtures cannot be related to concentrations. One limit loading level was tested.
Control
Six replicates (without test item) were tested under the same test conditions as the test vessels.
Test organisms
- Test organisms (species):
- Lemna gibba
- Details on test organisms:
- Test organism
Duckweed, Lemna gibba, Lemnaceae, Arales, Arecidae, Monocotyledonae
Young, rapidly growing plants without visible lesions or discolouration (chlorosis) were used for the test.
Reason for the selection of the test organism
According to the guideline, Lemna gibba is a suitable species because it is a representative of temperate areas commonly used for toxicity tests.
Origin
EUROFINS-GAB GMBH, Eutinger Str. 24, 75223 Niefern-Öschelbronn, Germany
Date of receipt
2008-02-26
Cultivation at test facility
The species is cultured in the test facility. Density is kept low to prevent conglomerates of plants on the surface. At least once per week, plants are transferred to freshly prepared growth medium. Growth media and culturing vessels are autoclaved before use to enable the breeding of axenic cultures.
Breeding vessels
Crystallisation dishes of glass, vol. 900 mL, filled with ca. 500 mL growth medium, covered with glass tops
Medium
20X-AAP-medium (Algal Assay Procedure medium),
pH-value 7.5 ± 0.1, see dilution water
Temperature 24 ± 2 °C
Light regime
Continuous fluorescent light, 1100 – 4440 lux
Acclimatization of the test system
The test system (the test organism) was held for 10 days under test conditions to acclimatize. These acclimatized plants were used in the test.
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 7 d
Test conditions
- Hardness:
- not measured
- Test temperature:
- see any other information on materials and methods
- pH:
- see any other information on materials and methods
- Dissolved oxygen:
- not measured
- Salinity:
- not measured
- Conductivity:
- not measured
- Details on test conditions:
- Test method
Static procedure
Test duration
7 days
Replicates
6 replicates per concentration level, 6 for the control
Test vessels/test volumes
Crystallisation dishes with a volume of 500 mL, covered with glass tops and filled with 200 mL test solution were used in the test. The test vessels were placed on a black non-reflective surface to avoid stray light.
Dilution water
20X-AAP-medium according to the guideline.
Composition of Dilution water
Component Concentration in stock solution [g/L] Concentration in prepared medium [mg/L]
NaNO3 26 510
MgCl2 6 H2O 12 240
CaCl2 2 H2O 4.4 90
MgSO4 7 H2O 15 290
K2HPO4 · 3 H2O 1.4 30
NaHCO3 15 300
H3BO3 0.19 3.7
MnCl2 4 H2O 0.42 8.3
FeCl3 6 H2O 0.16 3.2
Na2-EDTA · 2 H2O 0.30 6.0
ZnCl2 3.3 mg/L 66 µg/L
CoCl2 6 H2O 1.4 mg/L 29 µg/L
Na2MoO4 2 H2O 7.3 mg/L 145 µg/L
CuCl2 2 H2O 0.012 mg/L 0.24 µg/L
pH-value 7.5 ± 0.1
The pH of the test medium had to be 7.5 0.1 and was adjusted prior to testing with the addition of 1 N NaOH and HCl.
Application
Static with application of the test item at test start. At the start of the exposure, 3 uniform, healthy plants (colonies of 4 fronds each), were introduced into each test vessel containing the test media. The initial frond number per test vessel was 12. The initial numbers of colonies and fronds were the same in each test vessel.
Temperature (Target)
24 ± 2 °C
Light regime (Target)
Continuous, fluorescent light, 6500 to 10000 lux on the surface of the test medium (difference of light intensity at any measured incubation place < 15 % from the mean value)
Placement of the test vessels
A randomised placement of the test vessels was carried out.
Type and frequency of measurements
The numbers of plants and fronds were determined at the start and the end of the exposure. The number of fronds was determined every 2 - 3 days from each replicate of the control and the test concentrations. Every frond that visibly projected beyond the edge of a parent frond was counted as a separate frond. Fronds that lost their pigmentation were not counted.
Observations of frond size, appearance, indication of necrosis, chlorosis or gibbosity, colony break-up or loss of buoyancy, of root length and appearance, as well as of change in colour and destruction of roots, were made on every determination day and at the end of the exposure.
After 7 days, the determination of dry weight was carried out from 3 replicates per test concentration and 6 control replicates. Colonies from each test vessel were collected, rinsed with deionised water and then dried at 60 °C to a constant weight. Any root fragments were included. The dry weight was expressed to an accuracy of 0.1 mg.
The dry weight of the starting biomass was determined based on a sample of fronds (same number of fronds as in the test vessels) taken from the same batch used to inoculate the test vessels.
Physico-chemical Parameters
The pH-values were measured in the freshly prepared solutions before distribution into the replicates. The pH-values of the aged solution were measured from pooled replicates per concentration and control. The temperature of the medium in a surrogate vessel held under the same conditions in the growth room was recorded daily. The light intensity was measured prior to the start of the exposure at positions which had the same distance from the light source as the Lemna fronds. - Reference substance (positive control):
- yes
Results and discussion
Effect concentrationsopen allclose all
- Duration:
- 72 h
- Dose descriptor:
- other: NOEL
- Effect conc.:
- >= 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: frond number growth rate
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: frond number growth rate
- Duration:
- 72 h
- Dose descriptor:
- other: NOEL
- Effect conc.:
- >= 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Frond number yield
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: frond number yield
- Duration:
- 72 h
- Dose descriptor:
- other: NOEL
- Effect conc.:
- >= 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: dry weight growth rate
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: dry weight growth rate
- Duration:
- 72 h
- Dose descriptor:
- other: NOEL
- Effect conc.:
- >= 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: dry weight yield
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: dry weight yield
- Details on results:
- The environmental conditions (pH-value, room temperature, light intensity) were determined to be within the acceptable limits.
- Results with reference substance (positive control):
- The acute toxicity of 3,5-Dichlorophenol (SIGMA, batch number MKBZ0947V, purity 100.0 area %, CAS RN 591-35-5) to the monocotyledonous aquatic plant Lemna gibba was determined over a period of 7 days from 2017-10-06 to 2017-10-13 according to OECD Guideline 221. The plants used in the reference test were taken from the same laboratory culture as was used to determine the effects of Vat Black 9.
EC50-Values of the Reference Item
based on the nominal concentrations [mg/L], (0-7 days)
Current Study Valid Range (average ± 3 x SD)
Growth rate inhibition (number of fronds)
ErC50 6.76 5.71 ± 2.95
95% confidence interval 4.62 – 7.87
Yield inhibition (number of fronds)
EyC50 5.80 4.65 ± 2.96
95% confidence interval 4.50 – 7.05
Growth rate inhibition (dry weight)
ErdwC50 6.36 5.61 ± 2.76
95% confidence interval 5.14 – 7.12
Yield inhibition (dry weight)
EydwC50 5.39 4.67 ± 2.37
95% confidence interval 4.86 – 6.07
SD = standard deviation
The observed responses to the reference item were within the valid range, confirming the normal sensitivity of the test system used in the study with the test item. - Reported statistics and error estimates:
- EL-values and statistical analyses
EL-values were estimated empirically based on the results of the single treatment level (limit test design).
NOEL and LOEL values
NOEL/LOEL was determined by calculation of statistical significance of inhibition of growth rates and yield in comparison to the control using: One Way Analysis of Variance (ANOVA) and DUNNETT’s test as a standard. A normality test and an equal variance test were done first. The SHAPIRO-WILK-Test was used to test for normally distributed populations. The SPEARMAN rank test was used for equal variance test. P-values for both Normality and equal variance test are 0.05. The -value (acceptable probability of incorrectly concluding that there is a difference) is =0.05.
Software
All data will be computer generated and rounded for presentation from the full derived data. Consequently, if calculated manually based on the given data, minor variations may occur from these figures.
Calculations and graphics will be carried out using the following software
- Excel, MICROSOFT CORPORATION
- SigmaPlot, SPSS INC.
- GraphPad Prism, GRAPHPAD SOFTWARE, INC.
Any other information on results incl. tables
Frond Numbers
Nominal loading level |
Repl. No. |
Frond numbers per study day |
|||
0 days* |
2 days |
5 days |
7 days |
||
100 |
1 |
12 |
35 |
52 |
76 |
2 |
12 |
34 |
55 |
75 |
|
3 |
12 |
34 |
50 |
77 |
|
4 |
12 |
34 |
59 |
80 |
|
5 |
12 |
31 |
54 |
84 |
|
6 |
12 |
31 |
56 |
89 |
|
Mean |
12 |
33 |
54 |
80 |
|
Control |
1 |
12 |
33 |
53 |
74 |
2 |
12 |
38 |
59 |
92 |
|
3 |
12 |
39 |
57 |
107 |
|
4 |
12 |
34 |
53 |
95 |
|
5 |
12 |
36 |
56 |
100 |
|
6 |
12 |
39 |
50 |
72 |
|
Mean |
12 |
37 |
55 |
90 |
* = 3 colonies with 4 fronds each per replicate were inoculated at start of the exposure
Repl. No. = replicate number
Growth Rate and Yield Inhibition based on Fronds after 7 d
Statistically significant differences of growth rates and yield compared to control values are marked (+) and non-significant differences are marked (-).
Nominal loading level |
Repl. No. |
Average growth rate |
Inhibition of average growth rate |
Yield |
Inhibition of yield |
Doubling time |
|||||||||||
100 |
1 |
|
0.264 |
8 |
|
64 |
18 |
2.63 |
|||||||||
2 |
|
0.262 |
8 |
|
63 |
19 |
2.65 |
||||||||||
3 |
|
0.266 |
7 |
|
65 |
17 |
2.61 |
||||||||||
4 |
|
0.271 |
5 |
|
68 |
13 |
2.56 |
||||||||||
5 |
|
0.278 |
3 |
|
72 |
8 |
2.49 |
||||||||||
6 |
|
0.286 |
0 |
|
77 |
1 |
2.42 |
||||||||||
Mean |
(-) |
0.271 |
5 |
(-) |
68 |
13 |
2.56 |
||||||||||
Control |
1 |
|
0.260 |
|
|
62 |
|
2.67 |
|||||||||
2 |
|
0.291 |
|
|
80 |
|
2.38 |
||||||||||
3 |
|
0.313 |
|
|
95 |
|
2.22 |
||||||||||
4 |
|
0.296 |
|
|
83 |
|
2.35 |
||||||||||
5 |
|
0.303 |
|
|
88 |
|
2.29 |
||||||||||
6 |
|
0.256 |
|
|
60 |
|
2.71 |
||||||||||
Mean |
|
0.286 |
|
|
78 |
|
2.43 |
Repl. No. = replicate number
Specific Growth Rate and Yield Inhibition of Dry Weight after 7 d
Statistically significant differences of specific growth rates and yield compared to control values are marked (+) and non-significant differences are marked (-).
Nominal loading level |
Repl. No. |
Dry weight |
Specific dry weight growth rate |
Inhibition of specific dry weight growth rate |
Yield of dry weight |
Inhibition of yield dry weight |
||
100 |
1 |
13.6 |
|
0.281 |
11 |
|
11.7 |
25 |
2 |
14.6 |
|
0.291 |
8 |
|
12.7 |
19 |
|
3 |
16.8 |
|
0.311 |
2 |
|
14.9 |
5 |
|
4 |
17.6 |
|
0.318 |
0 |
|
15.7 |
0 |
|
5 |
15.0 |
|
0.295 |
7 |
|
13.1 |
17 |
|
6 |
17.6 |
|
0.318 |
0 |
|
15.7 |
0 |
|
Mean |
15.9 |
(-) |
0.303 |
5 |
(-) |
14.0 |
11 |
|
Control |
1 |
16.9 |
|
0.312 |
|
|
15.0 |
|
2 |
20.1 |
|
0.337 |
|
|
18.2 |
|
|
3 |
20.0 |
|
0.336 |
|
|
18.1 |
|
|
4 |
17.7 |
|
0.319 |
|
|
15.8 |
|
|
5 |
14.0 |
|
0.285 |
|
|
12.1 |
|
|
6 |
17.1 |
|
0.314 |
|
|
15.2 |
|
|
Mean |
17.6 |
|
0.317 |
|
|
15.7 |
|
The initial biomass dry weight was 1.9 mg per replicate.
Repl. No. = replicate number
Colony Number (Plants) on Days 0 and 7
Nominal loading level |
Replicate No. |
Colony number |
|||||
Day 0 |
Day 7 |
||||||
100 |
1 |
3 |
7 |
||||
2 |
3 |
7 |
|||||
3 |
3 |
7 |
|||||
4 |
3 |
7 |
|||||
5 |
3 |
8 |
|||||
6 |
3 |
8 |
|||||
Mean |
3 |
7 |
|||||
Control |
1 |
3 |
7 |
||||
2 |
3 |
7 |
|||||
3 |
3 |
8 |
|||||
4 |
3 |
8 |
|||||
5 |
3 |
8 |
|||||
6 |
3 |
7 |
|||||
Mean |
3 |
8 |
Further Observations on Days 3, 5 and 7
Nominal loading level |
Observations on day |
||||||
3 |
5 |
7 |
|||||
100 |
1 |
1 |
1 |
||||
Control |
1 |
1 |
1 |
Observations were made compared to the appearance of control colonies (plants) and test media
1 = no observedeffects
+ = slight effects
++ = medium effects
+++ = strong effects
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Conclusions:
- At the test item solution of the test item Vat Black 9 with a nominal concentration of 100 mg/L no effects were observed on the growth rate and yield (frond and dry weight) of Lemna gibba.
- Executive summary:
The effects of the test item Vat Black 9 on the growth of the monocotyledonous aquatic plant species Lemna gibba was determined according to the principles of OECD 221 and Council Regulation No. 761/2009 Method C.26 at the test facility from2017-10-05 to 2017-10-17, with the definitive exposure phase from 2017-10-06 to 2017-10-13.
Lemna gibba was exposed to the test item for 7 days under static conditions. Based on a preliminary test a limit concentration with a nominal loading of 100 mg/L was tested. Six replicates were investigated for the limit concentration and for the control. Frond numbers were assessed on days 0, 3, 5 and 7. Environmental parameters (light, pH and temperature) were within the acceptable limits. The validity criteria of the test guideline were fulfilled.
No analytical monitoring was carried out due to the low water solubility of the test item.
All effect values are given based on the nominal loading level.
NOEL-, LOEL-, EL-Values ofVat Black9after 7 Days of Exposure
(based on thenominal loading level[mg/L])
Frond number
Dry weight
Growth Rate Inhibition [mg/L]
NOEL
≥ 100
NOEL
≥ 100
LOEL
> 100
LOEL
> 100
ErL10
> 100
ErdwL10
> 100
ErL20
> 100
ErdwL20
> 100
ErL50
> 100
ErdwL50
> 100
Inhibition of Yield [mg/L]
NOEL
≥ 100
NOEL
≥ 100
LOEL
> 100
LOEL
> 100
EyL10
> 100
EydwL10
> 100
EyL20
> 100
EydwL20
> 100
EyL50
> 100
EydwL50
> 100
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