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EC number: 263-856-6 | CAS number: 63105-49-7
- 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
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- September 06th, 1995 to September 14th, 1995
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Remarks:
- The Prival modification was done to substitute the study from 1981
Cross-reference
- Reason / purpose for cross-reference:
- reference to other study
- Remarks:
- Standard plate incorporation test with Salmonella and e.coli tester strains with and without metabolic activation from induced rat liver
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 995
- Report date:
- 1995
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- OECD Guideline for testing of chemicals 471 Genetic Toxicology : Salmonella typhimurium, Reverse Mutation Assay, Adopted : May 26th, 1983
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OTS 798.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)
- Version / remarks:
- U.S. EPA: 798.5265 The Salmonella typhimurium reverse mutation assay Fed. Reg. 50, Subpart F, September 1985
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- EEC Directive 92/69, L 383 A, Annex B 14
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: M. J. Prival, V. D; Mitchell
- Version / remarks:
- M. J . Prival, V. D. Mitchell: Analysis of a method for testing azo dyes for mutagenicity in Salmonella typhimurium in the presence of flavine mononucleotide and hamster liver S-9. Mut. Res., 103- 106 (1982).
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Trisodium [5-acetamido-4-hydroxy-3-[[2-hydroxy-4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonato(5-)]cuprate(3-)
- EC Number:
- 263-856-6
- EC Name:
- Trisodium [5-acetamido-4-hydroxy-3-[[2-hydroxy-4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonato(5-)]cuprate(3-)
- Cas Number:
- 63105-49-7
- Molecular formula:
- C20H14CuN3O15S4.3Na C20H14CuN3Na3O15S4
- IUPAC Name:
- trisodium [5-acetamido-4-hydroxy-3-[[2-hydroxy-4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonato(5-)]cuprate(3-)
- Test material form:
- solid: particulate/powder
Constituent 1
Method
- Target gene:
- Histidine
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-mix fom uninduced hamster liver
- Test concentrations with justification for top dose:
- The first experiment was performed with all tester strains using three plates per dose to obtain information on mutagenicity and toxicity for calculating an appropriate dose range.
The test compound was tested at doses of 4 to 5000 microgram/plate and proved to be not toxic to the bacterial strains.
Therefore 5000 microgram/plate was chosen as the highest dose in the second experiment.
Test concentrations:
0, 4, 20, 100, 500, 2500 and 5000 μg/plate - Vehicle / solvent:
- On the day of the experiment the test compound was dissolved in aqua bidest. at appropriate concentrations.
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- congo red
- other: 2-Aminoanthracene (2-AA)
- Details on test system and experimental conditions:
- Preparation and storage of a liver homogenate fraction (S9)
Liver preparations were performed from the liver of non pre-treated Syrian hamsters.
The livers were removed from at least 5-6 male Syrian hamsters (7 -8 weeks old) at approx. 0 to 4 °C using cold sterile solutions and glassware, and were then pooled and washed in approx. 150 mM KCI (approximately 1 ml/g wet liver). The washed livers were cut into small pieces and homogenized in three volumes of KCI. The homogenate was centrifuged at approx. 9000 g for 10 minutes. The supernatant is the S9 fraction. This was divided into small portions, rapidly frozen and stored at approx. - 80 °C for not longer than six months.
Preparation of S9-mix
Sufficient S9 fraction was thawed immediately before each test at room temperature.
Three volumes of S9 fraction was mixed with 7 volumes of the S9 cofactor solution, which was kept on ice until used. This preparation is termed S9-mix. The concentrations of the different compounds in the S9-mix of the rat liver were:
8mM MgCI2
33mM KCI
20mM glucose-6-phosphate
2.8 units/ml glucose-6-phosphate dehydrogenase
4mM NADP+
2mM NADH
2mM FMN (Riboflavine-5'-phosphate-sodium-salt)
100mM phosphate buffer pH 7.4
According to the modification proposed by Prival using 30 minutes preincubation in the presence of 30 % (v/v) Syrian golden hamster S9-mix.
Bacteria
Bacteria were grown overnight in nutrient broth (25 g Oxoid Nutrient Broth No. 2 /liter) at approx. 37 °C. The amount of bacteria in the cell suspension was checked by nephelometry. Inoculation was performed with stock cultures which had been stored at approx. -80 °C. The compound was tested with the strains Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537. Identification of the different bacterial strains is performed periodically and all criteria for a valid assay were fulfilled as described.
Toxicity experiments and dose range finding
The first experiment was performed with all tester strains using three plates per dose to obtain information on mutagenicity and toxicity for calculating an appropriate dose range. A reduced rate of spontaneously occurring colonies and visible thinning of the bacterial lawn were used as toxicity indicators. Thinning of the bacterial lawn was evaluated microscopically.
In combination with the second experiment, toxicity testing was performed as follows:
0.1 ml of the different dilutions of the test compound were thoroughly mixed with 0.1 ml of 106 dilution of the overnight culture of TA 100 and plated with histidine and biotin rich top agar (3 plates per dose). The solvent control is compared with the number of colonies per plate in the presence of the test compound. Results are given as a ratio of these values (= surviving fraction).
Mutagenicity test
Two independent experiments were performed for each of the two protocols (Ames, Prival).
a) - with buffer and the strains TA 98, TA 100, TA 1535 and TA 1537 in a plate incorporation test without metabolic activation
Top agar was prepared for the Salmonella strains by mixing 100 ml agar (0.6% (w/v) agar, 0.5% (w/v) NaCI) with 10 ml of a 0.5 mM histidine-biotin solution.
The following ingredients were added (in the following order) to 2 ml of molten top agar at approx. 45 °C:
0.1 ml of an overnight nutrient broth culture of the bacterial tester strain
0.1 ml test compound solution
0.5 ml buffer
After mixing, the liquid was poured into a petri dish with minimal agar (1.5 % (w/v) agar, Vogel-Bonner E medium with 2 % (w/v) glucose). After incubation for approximately 48 hours at approx. 37 °C in the dark, colonies (his+ revertants) were counted.
b)- with 30% (v/v) Syrian golden hamster S9-mix and preincubation
0.1 ml test solution, 0.1 ml bacterial suspension and 0.5 ml S9-mix were incubated at approx. 30 °C for approx. 30 minutes. Subsequently, 2 ml of soft agar containing of 100 ml agar (0.6% (w/v) agar+ 0.5% (w/v) NaCI) and 10 ml amino-acid solution (minimal amino acid solution for the determination of mutants: 0.5 mM histidine + 0.5 mM biotin) was added. After mixing, the samples were poured on to the VogelBonner agar plates (minimal glucose agar plates) within approximately 30 seconds.
After incubation for 48 hours at 37 °C in the dark, colonies (his+ revertants) were counted. - Rationale for test conditions:
- In accordance with test guidelines.
- Evaluation criteria:
- A test compound is classified as mutagenic if it has either of the following effects:
a) a test compound produces at least a 2-fold increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control at complete bacterial background lawn
b) a test compound induces a dose-related increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control in at least two to three concentrations of the test compound at complete bacterial background lawn.
The test results must be reproducible.
Results and discussion
Test results
- Key result
- Species / strain:
- other: allstrains used
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Sterility checks and control plates
Sterility of S-9-mix and the test compound were indicated by the absence of contamination on the test material and S9-mix sterility check plates. Control plates (background control and positive controls) gave the expected number of colonies.
Solubility
The test compound did not precipitate on the plates up to the highest investigated dose of 5000 microgram/plate.
Toxicity
The test compound was tested at doses of 4 to 5000 microgram/plate and proved to be not toxic to the bacterial strains.
Therefore 5000 microgram/plate was chosen as the highest dose in the second experiment.
Mutagenicity
Experimental design
Remazol Brillantviolett 5R neu GT was tested for mutagenicity with Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537 in the absence and presence of a metabolic activation system. S9-mix from Syrian golden hamsters (30 % (v/v)) was used.
Mutation results
Ames-Test:
The test compound did not cause a significant increase in the number of revertant colonies with any of the tester strains in the absence of a metabolic activation system.
No dose dependent effect was obtained.
Prival-Test:
In the presence of hamster liver S9-mix (30% (v/v)) using the preincubation method according to Prival the test compound did not show any relevant increases in the number of revertant colonies under the experimental conditions described.
Two independent experiments for each of the two protocols (Ames, Prival) were performed.
Any other information on results incl. tables
CONTROLS – TEST 01
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVERTANTS/ PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 100 |
+S9 |
SOLVENT CONTROL |
|
|
|
|
|
||
|
158.7 |
30.4 |
|
|
181 |
124 |
171 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
152.3 |
26.3 |
1.0 |
|
132 |
182 |
143 |
||
POSITIVE CONTROLS |
|
2-AMINOANTHRACENE |
|
||||||
0.5 |
2303.0 |
111.6 |
14.5 |
|
2194 |
2417 |
2298 |
||
TA 100 |
-S9 |
SOLVENT CONTROLS |
|
|
|
|
|
||
|
126.0 |
15.6 |
|
|
117 |
144 |
117 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
135.0 |
24.6 |
1.1 |
|
125 |
163 |
117 |
||
POSITIVE CONTROLS |
|
SODIUM-AZIDE |
|
||||||
1.0 |
697.3 |
41.2 |
5.5 |
|
711 |
730 |
651 |
CONTROLS – TEST 01
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVERTANTS/ PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 98 |
+S9 |
SOLVENT CONTROL |
|
|
|
|
|
||
|
32.3 |
3.5 |
|
|
29 |
36 |
32 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
34.3 |
10.0 |
1.1 |
|
44 |
24 |
35 |
||
POSITIVE CONTROLS |
|
CONGORED |
|
||||||
500.0 |
258.3 |
9.5 |
8.0 |
|
249 |
258 |
268 |
||
TA 98 |
-S9 |
SOLVENT CONTROLS |
|
|
|
|
|
||
|
24.7 |
1.5 |
|
|
25 |
23 |
26 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
24.3 |
4.0 |
4.0 |
|
25 |
20 |
28 |
||
POSITIVE CONTROLS |
|
2-NITROFLUORENE |
|
||||||
2.5 |
831.7 |
71.8 |
33.7 |
|
910 |
816 |
769 |
TEST 01
THE EXPERIMENT WITH METABOLIC ACTIVATION WAS PERFORMED WITH 30% SYRIAN GOLDEN HAMSTER LIVER S9-MIX AND PREINCUBATION
ALL STERILITY CONTROL PLATES WERE STERILE
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVTERANTS /PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 100 |
+S9 |
0. 4. 20. 100. 500. 2500. 5000. |
158.7 157.7 171.0 158.0 175.7 173.7 194.7 |
30.4 23.4 18.3 6.1 23.4 32.6 5.0 |
1.0 1.1 1.0 1.1 1.1 1.2 |
|
181 153 151 161 149 136 200 |
124 183 175 162 193 193 190 |
171 137 187 151 185 192 194 |
TA 100 |
-S9 |
0. 4. 20. 100. 500. 2500. 5000. |
126.0 126.7 120.7 122.7 131.0 130.3 135.0 |
15.6 18.8 12.7 19.4 19.9 6.7 15.1 |
1.0 1.0 1.0 1.0 1.0 1.1 |
|
117 105 107 113 142 132 118 |
144 136 132 145 143 123 147 |
117 139 123 110 108 136 140 |
TA 98 |
+S9 |
0. 4. 20. 100. 500. 2500. 5000. |
32.3 36.0 37.3 30.0 33.3 24.0 24.7 |
3.5 7.0 9.1 4.6 10.4 4.0 3.1 |
1.1 1.2 0.9 1.0 0.7 0.8 |
|
29 39 44 34 45 28 24 |
36 41 41 31 25 24 28 |
32 28 27 25 30 20 22 |
TA 98 |
-S9 |
0. 4. 20. 100. 500. 2500. 5000. |
24.7 25.3 29.3 23.0 25.7 19.3 24.7 |
1.5 3.8 9.3 4.0 3.2 1.5 5.1 |
1.0 1.2 0.9 1.0 0.8 1.0 |
|
25 27 23 19 27 21 26 |
23 28 25 23 28 19 29 |
26 21 40 27 22 18 19 |
CONTROLS – TEST 01
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVERTANTS/ PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 1535 |
+S9 |
SOLVENT CONTROL |
|
|
|
|
|
||
|
14.7 |
4.5 |
|
|
19 |
15 |
10 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
10.0 |
2.0 |
0.7 |
|
8 |
12 |
10 |
||
POSITIVE CONTROLS |
|
2-AMINOANTHRACENE |
|
||||||
0.5 |
260.3 |
3.8 |
17.7 |
|
263 |
262 |
256 |
||
TA 1535 |
-S9 |
SOLVENT CONTROLS |
|
|
|
|
|
||
|
11.3 |
2.5 |
|
|
14 |
11 |
9 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
12.0 |
3.5 |
1.1 |
|
8 |
14 |
14 |
||
POSITIVE CONTROLS |
|
SODIUM-AZIDE |
|
||||||
1.0 |
380.3 |
27.8 |
33.7 |
|
360 |
412 |
369 |
CONTROLS – TEST 01
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVERTANTS/ PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 1537 |
+S9 |
SOLVENT CONTROL |
|
|
|
|
|
||
|
9.0 |
2.6 |
|
|
8 |
7 |
12 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
8.7 |
1.2 |
1.0 |
|
8 |
8 |
10 |
||
POSITIVE CONTROLS |
|
2-AMINOANTHRACENE |
|
||||||
2.5 |
230.0 |
6.6 |
25.6 |
|
223 |
236 |
231 |
||
TA 1537 |
-S9 |
SOLVENT CONTROLS |
|
|
|
|
|
||
|
7.7 |
1.5 |
|
|
9 |
6 |
8 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
6.3 |
0.6 |
0.8 |
|
6 |
6 |
7 |
||
POSITIVE CONTROLS |
|
9-AMINOACRIDINE |
|
||||||
50. |
130.0 |
5.6 |
16.9 |
|
124 |
135 |
131 |
TEST 01
THE EXPERIMENT WITH METABOLIC ACTIVATION WAS PERFORMED WITH 30% SYRIAN GOLDEN HAMSTER LIVER S9-MIX AND PREINCUBATION
ALL STERILITY CONTROL PLATES WERE STERILE
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVTERANTS /PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 1535 |
+S9 |
0. 4. 20. 100. 500. 2500. 5000. |
14.7 11.3 12.0 8.3 10.3 11.0 10.0 |
4.5 3.5 2.0 0.6 1.5 2.6 2.6 |
0.8 0.8 0.6 0.7 0.7 0.7 |
|
19 15 10 9 12 10 13 |
15 8 12 8 10 9 9 |
10 11 14 8 9 14 8 |
TA 1535 |
-S9 |
0. 4. 20. 100. 500. 2500. 5000. |
11.3 10.0 11.0 11.0 10.0 9.7 8.7 |
2.5 2.0 2.6 2.6 2.0 2.1 1.2 |
0.9 1.0 1.0 0.9 0.9 0.8 |
|
14 12 12 9 8 9 8 |
11 10 13 10 12 12 10 |
9 8 8 14 10 8 8 |
TA 1537 |
+S9 |
0. 4. 20. 100. 500. 2500. 5000. |
9.0 9.0 9.0 7.3 8.7 9.3 6.7 |
2.6 1.0 1.0 2.3 0.6 1.2 1.2 |
1.0 1.0 0.8 1.0 1.0 0.7 |
|
8 9 9 6 9 10 8 |
7 10 8 6 8 10 6 |
12 8 10 10 9 8 6 |
TA 1537 |
-S9 |
0. 4. 20. 100. 500. 2500. 5000. |
7.7 8.7 8.7 8.7 7.3 9.7 9.7 |
1.5 1.2 1.2 2.3 2.3 9.7 9.7 |
1.1 1.1 1.1 0.9 1.3 1.3 |
|
9 8 10 10 6 10 9 |
6 8 8 10 6 10 10 |
8 10 8 6 10 9 10 |
CONTROLS – TEST 02
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVERTANTS/ PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 100 |
+S9 |
SOLVENT CONTROL |
|
|
|
|
|
||
|
149.0 |
11.3 |
|
|
136 |
156 |
155 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
156.7 |
6.0 |
1.1 |
|
151 |
156 |
163 |
||
POSITIVE CONTROLS |
|
2-AMINOANTHRACENE |
|
||||||
0.5 |
2286.3 |
88.3 |
15.3 |
|
2359 |
2188 |
2312 |
||
TA 100 |
-S9 |
SOLVENT CONTROLS |
|
|
|
|
|
||
|
110.3 |
6.7 |
|
|
107 |
118 |
106 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
114.0 |
12.1 |
1.0 |
|
128 |
107 |
107 |
||
POSITIVE CONTROLS |
|
SODIUM-AZIDE |
|
||||||
1.0 |
703.0 |
25.9 |
6.4 |
|
731 |
680 |
698 |
CONTROLS – TEST 02
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVERTANTS/ PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 1535 |
+S9 |
SOLVENT CONTROL |
|
|
|
|
|
||
|
10.3 |
0.6 |
|
|
10 |
11 |
10 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
11.7 |
2.1 |
1.1 |
|
14 |
10 |
11 |
||
POSITIVE CONTROLS |
|
2-AMINOANTHRACENE |
|
||||||
0.5 |
254.7 |
5.5 |
24.7 |
|
249 |
255 |
260 |
||
TA 1535 |
-S9 |
SOLVENT CONTROLS |
|
|
|
|
|
||
|
10.3 |
1.2 |
|
|
9 |
11 |
11 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
10.3 |
0.6 |
1.0 |
|
11 |
10 |
10 |
||
POSITIVE CONTROLS |
|
SODIUM-AZIDE |
|
||||||
1.0 |
344.7 |
30.2 |
33.5 |
|
322 |
379 |
333 |
CONTROLS – TEST 02
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVERTANTS/ PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 1537 |
+S9 |
SOLVENT CONTROL |
|
|
|
|
|
||
|
9.3 |
1.5 |
|
|
8 |
9 |
11 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
8.0 |
2.0 |
0.9 |
|
8 |
6 |
10 |
||
POSITIVE CONTROLS |
|
2-AMINOANTHRACENE |
|
||||||
2.5 |
227.3 |
7.5 |
24.4 |
|
236 |
223 |
223 |
||
TA15 37 |
-S9 |
SOLVENT CONTROLS |
|
|
|
|
|
||
|
7.7 |
1.5 |
|
|
6 |
8 |
9 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
8.0 |
0.0 |
1.0 |
|
8 |
8 |
8 |
||
POSITIVE CONTROLS |
|
9-AMINOACRIDINE |
|
||||||
50. |
127.7 |
9.0 |
16.6 |
|
123 |
122 |
138 |
CONTROLS – TEST 02
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVERTANTS/ PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 98 |
+S9 |
SOLVENT CONTROL |
|
|
|
|
|
||
|
35.0 |
6.9 |
|
|
39 |
27 |
39 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
34.0 |
11.1 |
1.0 |
|
44 |
22 |
36 |
||
POSITIVE CONTROLS |
|
COGORED |
|
||||||
500.0 |
259.3 |
38.7 |
7.4 |
|
215 |
277 |
286 |
||
TA 98 |
-S9 |
SOLVENT CONTROLS |
|
|
|
|
|
||
|
23.7 |
3.8 |
|
|
28 |
22 |
21 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
27.3 |
2.1 |
1.2 |
|
29 |
28 |
25 |
||
POSITIVE CONTROLS |
|
2-NITROFLUORENE |
|
||||||
2.5 |
1098.0 |
135.0 |
46.3 |
|
961 |
1102 |
1231 |
TEST 02
THE EXPERIMENT WITH METABOLIC ACTIVATION WAS PERFORMED WITH 30% SYRIAN GOLDEN HAMSTER LIVER S9-MIX AND PREINCUBATION
ALL STERILITY CONTROL PLATES WERE STERILE
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVTERANTS /PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 100 |
+S9 |
0. 4. 20. 100. 500. 2500. 5000. |
149.0 154.7 157.0 155.7 168.7 188.3 179.7 |
11.3 26.1 7.8 11.1 4.6 8.5 36.0 |
1.0 1.1 1.0 1.1 1.3 1.2 |
|
136 161 148 144 174 185 143 |
156 177 161 66 166 182 181 |
155 126 162 157 166 198 215 |
TA 100 |
-S9 |
0. 4. 20. 100. 500. 2500. 5000. |
110.3 116.7 116.7 126.3 117.7 136.3 139.7 |
6.7 15.1 11.8 16.3 9.5 23.0 6.7 |
1.1 1.1 1.1 1.1 1.2 1.3 |
|
107 106 103 123 118 135 143 |
118 110 123 144 127 160 144 |
106 134 124 112 108 114 132 |
TA 1535 |
+S9 |
0. 4. 20. 100. 500. 2500. 5000. |
10.3 11.0 12.0 12.0 12.3 11.7 11.3 |
0.6 2.6 0.0 2.0 2.9 0.6 2.5 |
1.1 1.2 1.2 1.2 1.1 1.1 |
|
10 10 12 12 9 12 14 |
11 14 12 10 14 11 9 |
10 9 12 14 14 12 11 |
TA 1535 |
-S9 |
0. 4. 20. 100. 500. 2500. 5000. |
10.3 8.3 12.7 13.0 10.0 12.3 10.7 |
1.2 0.6 2.3 1.7 1.7 4.5 2.9 |
0.8 1.2 1.3 1.0 1.2 1.0 |
|
9 9 10 14 9 12 9 |
11 8 14 11 12 8 14 |
11 8 14 14 9 17 9 |
TEST 02
THE EXPERIMENT WITH METABOLIC ACTIVATION WAS PERFORMED WITH 30% SYRIAN GOLDEN HAMSTER LIVER S9-MIX AND PREINCUBATION
ALL STERILITY CONTROL PLATES WERE STERILE
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVTERANTS /PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 1537 |
+S9 |
0. 4. 20. 100. 500. 2500. 5000. |
9.3 9.3 8.7 8.3 7.7 8.7 7.0 |
1.5 2.3 1.2 1.5 2.1 1.2 1.0 |
1.0 0.9 0.9 0.8 0.9 0.8 |
|
8 12 8 10 7 8 6 |
9 8 10 7 10 10 8 |
11 8 8 8 6 8 7 |
TA 1537 |
-S9 |
0. 4. 20. 100. 500. 2500. 5000. |
7.7 8.3 10.0 8.3 9.3 9.0 7.3 |
1.5 1.2 0.0 0.6 0.6 1.0 1.5 |
1.1 1.3 1.1 1.2 1.2 0.9 |
|
6 9 10 8 9 9 6 |
8 7 10 8 9 10 7 |
9 9 10 9 10 8 9 |
TA 98 |
+S9 |
0. 4. 20. 100. 500. 2500. 5000. |
35.0 30.7 29.0 29.7 29.7 28.0 23.0 |
6.9 2.5 2.6 6.5 8.6 6.2 4.4 |
0.9 0.8 0.8 0.8 0.8 0.7 |
|
39 28 30 30 39 23 21 |
27 33 26 23 22 35 20 |
39 31 31 36 28 26 28 |
TA 98 |
-S9 |
0. 4. 20. 100. 500. 2500. 5000. |
23.7 25.0 24.3 25.0 21.0 22.7 23.3 |
3.8 4.4 2.9 1.7 1.0 2.1 3.2 |
1.1 1.0 1.1 0.9 1.0 1.0 |
|
28 27 26 27 22 25 27 |
22 20 21 24 20 21 22 |
21 28 26 24 21 22 21 |
CONTROLS – TEST 03
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVERTANTS/ PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 100 D |
+S9 |
SOLVENT CONTROL |
|
|
|
|
|
||
|
201.3 |
27.5 |
|
|
201 |
229 |
174 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
176.3 |
24.1 |
0.9 |
|
156 |
170 |
203 |
||
POSITIVE CONTROLS |
|
NOT TESTED |
|
||||||
|
|
*** NO RESULTS FOR THIS STRAIN *** |
|
||||||
TA 100 D |
-S9 |
SOLVENT CONTROLS |
|
|
|
|
|
||
|
162.3 |
5.5 |
|
|
162 |
168 |
157 |
||
NEGATIVE CONTROLS |
|
|
|
|
|
||||
|
148.0 |
26.0 |
0.9 |
|
132 |
134 |
178 |
||
POSITIVE CONTROLS |
|
NOT TESTED |
|
||||||
|
|
*** NO RESULTS FOR THIS STRAIN *** |
|
TEST 03
THE EXPERIMENT WITH METABOLIC ACTIVATION WAS PERFORMED WITH 30% SYRIAN GOLDEN HAMSTER LIVER S9-MIX AND PREINCUBATION
STRAIN |
DOSE LEVELS (μg/plate) |
MEAN |
STANDARD DEVIATION |
RATIO: TEST/ CONTROL |
BACTERIAL LAWN |
NO REVTERANTS /PLATE |
|||
PLATE 1 |
PLATE 2 |
PLATE 3 |
|||||||
TA 100 D |
+S9 |
0. 4. 20. 100. 500. 2500. 5000. |
201.3 179.3 185.3 154.7 178.3 197.3 195.0 |
27.5 6.0 23.2 19.7 14.6 21.2 4.4 |
0.9 0.9 0.8 0.9 1.0 1.0 |
|
201 180 210 167 163 178 198 |
229 173 182 132 192 194 197 |
174 185 164 165 180 220 190 |
TA 100 D |
-S9 |
0. 4. 20. 100. 500. 2500. 5000. |
162.3 135.3 127.7 137.3 132.3 135.0 126.7 |
5.5 30.8 23.3 18.6 22.7 30.6 19.5 |
0.8 0.8 0.8 0.8 0.8 0.8 |
|
162 125 101 122 108 122 118 |
168 111 138 132 136 113 113 |
157 170 144 158 153 170 149 |
Applicant's summary and conclusion
- Conclusions:
- The results lead to the conclusion that Remazol Brillantviolett 5R neu GT is not mutagenic in the absence of a metabolic activation system using the standard Ames Test procedure. Also in the presence of hamster liver S9-mix (30 % (v/v)) and preincubation the test compound did not induce a significant increase in the number of revertant colonies. The substance did not cause any cytotoxic effects.
- Executive summary:
Remazol Brillantviolett 5R neu GT was tested for mutagenicity with the strains TA 100, TA 1535, TA 1537 and TA 98 of Salmonella typhimurium.
The mutagenicity studies were conducted in the standard plate test (Ames Test) and in a modified preincubation test (Prival Test). The studies were performed in the absence and in the presence of a metabolising system derived from hamster liver homogenate. The test compound was dissolved in aqua bidest. and a dose range of 6 different doses from 4 microgram/plate to 5000 microgram/plate was used.
Control plates without mutagen showed that the number of spontaneous revertant colonies was similar to that described in the literature. All the positive control compounds gave the expected increase in the number of revertant colonies.
Toxicity: The test compound proved to be not toxic to the bacterial strains.
5000 microgram/plate was chosen as top dose level for the mutagenicity study.
a) Ames Test:
Mutagenicity: In the absence of the metabolic activation system Remazol Brillantviolett 5R neu GT did not show a dose dependent increase in the number of revertants in any of the bacterial strains.
b) Prival Test:
In the presence of hamster liver S9-mix (30 % (v/v)) using the preincubation method according to Prival Remazol Brillantviolett 5R neu GT did not induce a significant increase in the number of revertant colonies with any of the tester strains.
Summarising, it can be stated that Remazol Brillantviolett 5R neu GT is not mutagenic in the standard plate test (Ames Test) and in the preincubation method according to Prival.
The test substance is not classified according to CLP criteria.
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