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EC number: 610-202-6 | CAS number: 446299-80-5
- 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:
- From 08-08-2022 to 26-08-2022
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 022
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- N-(5-bromo-3-methylpyridin-2-yl)-N-methylbenzamide
- EC Number:
- 610-202-6
- Cas Number:
- 446299-80-5
- Molecular formula:
- C14H13BrN2O
- IUPAC Name:
- N-(5-bromo-3-methylpyridin-2-yl)-N-methylbenzamide
- Reference substance name:
- N-(3-bromo-5-methyl pyridine-2-yl)-N-methylbenzamide
- Molecular formula:
- C14H13BrN2O
- IUPAC Name:
- N-(3-bromo-5-methyl pyridine-2-yl)-N-methylbenzamide
- Reference substance name:
- Benzoic acid
- EC Number:
- 200-618-2
- EC Name:
- Benzoic acid
- Cas Number:
- 65-85-0
- Molecular formula:
- C7H6O2
- IUPAC Name:
- Benzoic acid
- Reference substance name:
- Unknown impurities
- IUPAC Name:
- Unknown impurities
- Reference substance name:
- Water
- EC Number:
- 231-791-2
- EC Name:
- Water
- Cas Number:
- 7732-18-5
- Molecular formula:
- H2O
- IUPAC Name:
- water
- Test material form:
- solid: particulate/powder
Constituent 1
impurity 1
impurity 2
impurity 3
impurity 4
Method
- Target gene:
- his D (S. typhimurium TA 98); his C (S. typhimurium TA 1537); his G (S. typhimurium TA 100 and TA1535); tryp E (E. coli WP2 uvrA pKM101)
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Species / strain / cell type:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Metabolic activation system:
- Preparation os the metabolic activation system:
- Obtention of S9: S9 fraction prepared from Sprague Dawley rat liver homogenate, provided by MOLTOXTM (POB Box 1189 - 157 Industrial Park Dr - Boone, NC 28607 - USA) (S9 mOLTOX-11-101.5-447 VALIDATED ON 11.10.2021 - EXPIRY DATE: 13.07.2023).
- Preparation of S9 mix 10%: 10% S9 fraction, 8 mM MgCL2-6H2O, 33 mM KCl, 5 mM Glucose-6-Phosphate Na2, 4 mM NADP Na2 and 0.1 M Phosphate buffer pH 7.4.
- preliminary cytotoxicity testing: Sterility test: test item and the corresponding dilutions were added to 2 mL of top agar at 45ºC, homogenized and poured on the agar (20 ml) onto a Petri plate in three plates. Plates were incubated for 48 - 72 hours at 37°C. - Test concentrations with justification for top dose:
- Initial test concentrations: 50, 150, 500, 1500 and 5000 µg/plate.
No toxicity was found for doses up to 5000 μg/plate. That's why the test item was tested at concentration of 5000 μg/plate (maximum one). - Vehicle / solvent:
- - Vehicle/solvent: Dimethyl sulfoxide (DMSO)
- Justification for choice of solvent/vehicle: the test item was found soluble in DMSO at the highest tested concentration.
- Details on test system and experimental conditions:
- PRELIMINARY CYTOTOXICITY TEST:
In a test tube 0.1 mL of the bacterial suspension (1-9 X 10^3 bacteria /mL) and 0.1 mL of the stock solution and dilutions were successively added to 2 mL of top agar at 45ºC, containing 10 % (v/v) of a solution of L-Histidine-D-Biotine (2.5 mM). After homogenization, the content of the tube was poured onto a Petri plate (90 mm in diameter) containing minimal agar (20 mL). 3 plates per concentration were incubated for 48-72 h at 37ºC, and the colonies counted. A negative control containing the blank alone was run in parallel.
In case of bacteriostatic activity is detected, the highest concentration to be retained is that exhibiting a bacteriostatic activity of 75% or less. The precipitate, if present, should not interfere with the scoring.
METHOD:
1. Without metabolic activation: Salmonella Typhimurium strains: 0.1 mL of the bacterial suspension containing 1-9 x10^9 bacteria/mL and 50µl of each dilution of the original solution and 0.5 mL of sterile phosphate buffer are successively added to 2 mL of overlay agar maintained super cooled at 45ºC containing 10% (v/v) of a L-Histidine-D-Biotine solution (0.5 mM).
For Escherichia coli strain containing 5% (v/v) of nutrient broth nº2 to which are added 5 μL of a L-Tryptophane solution at 2 mg/mL.
With metabolic activation, is similar to the described above, except that, 500 μL of S9-mix fraction is quickly added, before pouring the mixture onto the plates. After a 48-72 hour incubation period at 37ºC, revertant colonies are counted in each plate.
2. Pre-incubation: The test item solution with the test strain, and 500 μL of S9-mix fraction are preincubated with shaking for 30 min., at 37ºC prior to mixing with the overlay agar and pouring onto the minimal agar plate. After a 48-72 hour incubation period at 37ºC, revertant colonies are counted in each plate.
DATA PRESENTATION
After a 48-72-hour incubation period at 37ºC, revertant colonies were manually counted in each plate. The following ratio was calculated per plate: R = Number of revertant colonies in the presence of the test item / Number of revertant colonies in the absence of the test item. - Evaluation criteria:
- The following validity criteria were checked to validate each experiment:
- the bacteriostatic activity of the highest concentration tested shall be equal to or less than 75%,
- the spontaneous reversion rate of the absolute negative control shall comply with the historical values of the laboratory,
- in presence of the solvent the spontaneous reversion rate shall comply with the historial values of the absolute negative control of the laboratory
- the mean number of revertant colonies obtained for each strain and the corresponding positive control, with and/or without metabolic activation shall comply with the historical values of the laboratory.
- negative and positive values should not show significant difference with the historical values of the laboratory (± standard deviations).
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 98
- 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
- Key result
- Species / strain:
- S. typhimurium TA 100
- 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
- Key result
- Species / strain:
- S. typhimurium TA 1535
- 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
- Key result
- Species / strain:
- S. typhimurium TA 1537
- 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
- Key result
- Species / strain:
- E. coli WP2 uvr A
- 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
Any other information on results incl. tables
Table 3. Sterility control.
Serie |
Doses |
Colony number/plate | ||
Control n° 1 | 1 | 2 | 3 | |
Solution of
T-A10 BATCH: M22154C
(Identification code :PH-22/0484) | 5000 µg /plate | 0 | 0 | 0 |
1500 µg /plate | 0 | 0 | 0 | |
500 µg /plate | 0 | 0 | 0 | |
150 µg /plate | 0 | 0 | 0 | |
50 µg /plate | 0 | 0 | 0 | |
S9-mix | 500 µL/plate | 0 | 0 | 0 |
Control n° 2 | 1 | 2 | 3 | |
Solution of
T-A10 BATCH: M22154C
(Identification code :PH-22/0484) | 5000 µg /plate | 0 | 0 | 0 |
1500 µg /plate | 0 | 0 | 0 | |
500 µg /plate | 0 | 0 | 0 | |
150 µg /plate | 0 | 0 | 0 | |
50 µg /plate | 0 | 0 | 0 | |
S9-mix | 500 µL/plate | 0 | 0 | 0 |
Table 4. Bacteriostatic activity controls.
| Doses (/plate) | |||||||||||||||||||
0 (negative control) |
DMSO control |
50 µg |
150 µg |
500 µg |
1 500 µg |
5 000 µg | ||||||||||||||
| N1 |
| 900 |
|
| 864 |
|
| 869 |
|
| 831 |
|
| 867 |
| 860 |
| 184 |
|
Solution of | N2 |
| 885 |
|
| 900 |
|
| 896 |
|
| 854 |
|
| 890 |
| 894 |
| 264 |
|
T-A10 BATCH: M22154C | N3 |
| 880 |
|
| 844 |
|
| 854 |
|
| 876 |
|
| 899 |
| 781 |
| 254 |
|
| N | 888 | ± | 10 | 869 | ± | 28 | 873 | ± | 21 | 854 | ± | 23 | 885 | ± | 17 | 845±58 | 234 | ± | 44 |
| % |
| - |
|
| 98% |
|
| 98% |
|
| 96% |
|
| 100% |
| 95% |
| 26% |
|
| Doses (/plate) | |||||||||||||||||||
0 (negative control) |
DMSO |
50 µg |
150 µg |
500 µg |
1 500 µg |
5 000 µg | ||||||||||||||
| N1 |
| 684 |
|
| 701 |
|
| 702 |
|
| 698 |
|
| 701 |
| 645 |
| 210 |
|
Solution of | N2 |
| 676 |
|
| 689 |
|
| 698 |
|
| 685 |
|
| 626 |
| 621 |
| 168 |
|
T-A10 BATCH: M22154C | N3 |
| 699 |
|
| 677 |
|
| 652 |
|
| 674 |
|
| 691 |
| 632 |
| 174 |
|
| N | 686 | ± | 12 | 689 | ± | 12 | 684 | ± | 28 | 686 | ± | 12 | 676 | ± | 44 | 633±12 | 788 | ± | 37 |
| % |
| - |
|
| 100% |
|
| 100% |
|
| 100% |
|
| 98% |
| 92% |
| 27% |
|
Table 5. Result tables.
TA1535 Assay n°1 – without metabolic activation (-S9-mix) | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 7 | 13 | 9 | 9.67 | 3.06 | _ |
Positive control solvent | 5 µL | 11 | 19 | 12 | 14.00 | 4.36 | _ |
Positive control : Sodium azide | 5 µg in 5 µL |
462 |
513 |
577 |
517.33 |
57.62 |
36.95 |
Vehicle | 50µL | 11 | 12 | 15 | 12.67 | 2.08 | _ |
| 5000 µg | 12 | 13 | 11 | 9.00 | 1.00 | 0.95 |
Solution of | 1500 µg | 13 | 15 | 16 | 10.33 | 1.53 | 1.16 |
T-A10 BATCH: M22154C | 500 µg | 17 | 12 | 12 | 12.33 | 2.89 | 1.08 |
| 150 µg | 12 | 20 | 15 | 14.33 | 4.04 | 1.24 |
| 50 µg | 17 | 9 | 8 | 11.33 | 4.93 | 0.89 |
TA1535 Assay n°1 – with metabolic activation (10 % S9-mix) – without pre-incubation | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 9 | 12 | 14 | 11.67 | 2.52 | _ |
Positive control solvent | 20 µL | 11 | 13 | 8 | 10.67 | 2.52 | _ |
Positive control : 2-Anthramine | 2 µg in 20 µL |
164 |
125 |
112 |
133.67 |
27.06 |
12.53 |
Vehicle | 50µL | 7 | 10 | 10 | 9.00 | 1.73 | _ |
| 5000 µg | 14 | 8 | 11 | 11.00 | 3.00 | 1.22 |
Solution of | 1500 µg | 11 | 18 | 12 | 13.67 | 3.79 | 1.52 |
T-A10 BATCH: M22154C | 500 µg | 13 | 10 | 16 | 13.00 | 3.00 | 1.44 |
| 150 µg | 12 | 15 | 20 | 15.67 | 4.04 | 1.74 |
| 50 µg | 9 | 20 | 14 | 14.33 | 5.51 | 1.59 |
TA1535 Assay n°2 – without metabolic activation (-S9 mix) | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 15 | 11 | 12 | 12.67 | 2.08 | _ |
Positive control solvent | 5 µL | 7 | 12 | 8 | 9.00 | 2.65 | _ |
Positive control : Sodium azide | 5 µg in 5 µL |
807 |
960 |
855 |
874.00 |
78.25 |
97.11 |
Vehicle | 50µL | 6 | 9 | 10 | 8.33 | 2.08 | _ |
| 5000 µg | 7 | 17 | 8 | 10.67 | 5,51 | 1.28 |
Solution of | 1500 µg | 17 | 15 | 11 | 14.33 | 3.06 | 1.72 |
T-A10 BATCH: M22154C | 500 µg | 9 | 6 | 10 | 8.33 | 1,53 | 1.00 |
| 150 µg | 7 | 10 | 8 | 8.33 | 4,16 | 1.00 |
| 50 µg | 9 | 9 | 13 | 10.33 | 4,58 | 1.24 |
TA1535 Assay n°2 – with metabolic activation (10 % S9-mix) – with pre-incubation | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 9 | 11 | 8 | 9.33 | 1.53 | _ |
Positive control solvent | 10 µL | 11 | 10 | 7 | 9.33 | 2.08 | _ |
Positive control : 2-Anthramine | 1 µg in 10 µL |
108 |
83 |
73 |
88.00 |
18.03 |
9.43 |
Vehicle | 50µL | 11 | 6 | 12 | 9.67 | 3.21 | _ |
| 5000 µg | 9 | 13 | 11 | 11.00 | 2.00 | 1.14 |
Solution of | 1500 µg | 14 | 16 | 12 | 14.00 | 2.00 | 1.45 |
T-A10 BATCH: M22154C | 500 µg | 17 | 14 | 19 | 16.67 | 2.52 | 1.72 |
| 150 µg | 10 | 17 | 18 | 15.00 | 4.36 | 1.55 |
| 50 µg | 17 | 12 | 13 | 14.00 | 2.65 | 1.45 |
TA1537 Assay n°1 – without metabolic activation (-S9-mix) | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 6 | 7 | 6 | 6.33 | 0.58 | _ |
Positive control solvent | 20 µL | 10 | 4 | 7 | 7.00 | 3.00 | _ |
Positive control : 9-Aminoacridine | 50 µg |
645 |
793 |
845 |
761.00 |
103.77 |
108.71 |
Vehicle | 50µL | 11 | 5 | 5 | 7.00 | 3.46 | _ |
| 5000 µg | 6 | 3 | 6 | 5.00 | 1.73 | 0.71 |
Solution of | 1500 µg | 6 | 8 | 7 | 7.00 | 1.00 | 1.00 |
T-A10 BATCH: M22154C | 500 µg | 5 | 6 | 4 | 5.00 | 1.00 | 0.71 |
| 150 µg | 8 | 7 | 3 | 6.00 | 2.65 | 0.86 |
| 50 µg | 4 | 10 | 7 | 7.00 | 3.00 | 1.700 |
TA1537 Assay n°1 – with metabolic activation (10 % S9-mix) – without pre-incubation | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 12 | 7 | 16 | 11.67 | 4.51 | _ |
Positive control solvent | 20 µL | 10 | 10 | 11 | 10.33 | 0.58 | _ |
Positive control : 2-Anthramine | 50 µL |
38 |
40 |
39 |
39.00 |
1.00 |
3.77 |
Vehicle | 50µL | 10 | 8 | 16 | 11.33 | 4.16 | _ |
| 5000 µg | 7 | 5 | 8 | 6.67 | 1.53 | 0.59 |
Solution of | 1500 µg | 18 | 16 | 11 | 15.00 | 3.61 | 1.32 |
T-A10 BATCH: M22154C | 500 µg | 9 | 10 | 12 | 10.33 | 1.53 | 0.91 |
| 150 µg | 6 | 12 | 18 | 12.00 | 6.00 | 1.06 |
| 50 µg | 18 | 13 | 10 | 13.67 | 4.04 | 1.21 |
TA1537 Assay n°2 – without metabolic activation (-S9-mix) | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 3 | 13 | 5 | 7.00 | 5.29 | _ |
Positive control solvent | 20 µL | 4 | 10 | 7 | 7.00 | 3.00 | _ |
Positive control : 9-Aminoacridine | 50 µg
|
878 |
1293 |
666 |
945.67 |
318.93 |
135.10 |
Vehicle | 50µL | 6 | 5 | 8 | 6.33 | 1.53 | _ |
| 5000 µg | 1 | 1 | 2 | 1.33 | 0.58 | 0.21 |
Solution of | 1500 µg | 2 | 8 | 5 | 5.00 | 3.00 | 0.79 |
T-A10 BATCH: M22154C | 500 µg | 8 | 6 | 4 | 6.00 | 2.00 | 0.95 |
| 150 µg | 10 | 9 | 8 | 9.00 | 1.00 | 1.42 |
| 50 µg | 3 | 7 | 7 | 5.67 | 2.31 | 0.89 |
TA1537 Assay n°2 – with metabolic activation (10% S9-mix) – with pre-incubation | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 5 | 8 | 8 | 7.00 | 1.73 | _ |
Positive control solvent | 10 µL | 12 | 8 | 10 | 10.00 | 2.500 | _ |
Positive control : 2-Anthramine | 50 µL |
26 |
28 |
25 |
26.33 |
1.53 |
2.63 |
Vehicle | 50µL | 6 | 4 | 8 | 6.00 | 2.00 | _ |
| 5000 µg | 1 | 6 | 2 | 3.00 | 2.65 | 0.50 |
Solution of | 1500 µg | 4 | 8 | 10 | 7.33 | 3.06 | 1.22 |
T-A10 BATCH: M22154C | 500 µg | 8 | 12 | 10 | 10.00 | 2.00 | 1.67 |
| 150 µg | 4 | 10 | 11 | 8.33 | 3.79 | 1.39 |
| 50 µg | 8 | 12 | 9 | 9.67 | 2.08 | 1.61 |
TA98 Assay n°1 – without metabolic activation (-S9-mix) | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 23 | 23 | 18 | 21.33 | 2.89 | _ |
Positive control solvent | 20 µL | 15 | 19 | 20 | 18.00 | 2.65 | _ |
Positive control : 2-Nitrofluorene | 50 µL |
427 |
336 |
426 |
396.33 |
52.25 |
22.02 |
Vehicle | 50µL | 19 | 18 | 23 | 20.00 | 2.65 | _ |
| 5000 µg | 28 | 24 | 14 | 22.00 | 7.21 | 1.10 |
Solution of | 1500 µg | 18 | 21 | 18 | 19.00 | 1.73 | 0.95 |
T-A10 BATCH: M22154C | 500 µg | 26 | 27 | 26 | 26.33 | 0.58 | 1.32 |
| 150 µg | 18 | 15 | 29 | 20.67 | 7.37 | 1.03 |
| 50 µg | 12 | 18 | 20 | 16.67 | 4.16 | 0.83 |
TA98 Assay n°1 – with metabolic activation (10 % S9-mix) – without pre-incubation | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 26 | 25 | 25 | 25.33 | 0.58 | _ |
Positive control solvent | 20 µL | 31 | 25 | 23 | 26.33 | 4.16 | _ |
Positive control : 2-Anthramine | 50 µL |
407 |
321 |
293 |
340.33 |
59.41 |
12.92 |
Vehicle | 50µL | 21 | 36 | 24 | 27.00 | 7.94 | _ |
| 5000 µg | 24 | 27 | 14 | 21.67 | 6.81 | 0.80 |
Solution of | 1500 µg | 33 | 31 | 34 | 32.67 | 1.53 | 1.21 |
T-A10 BATCH: M22154C | 500 µg | 34 | 26 | 27 | 29.00 | 4.36 | 1.07 |
| 150 µg | 31 | 34 | 38 | 34.33 | 3.51 | 1.27 |
| 50 µg | 30 | 36 | 26 | 30.67 | 5.03 | 1.14 |
TA98 Assay n°2 – without metabolic activation (-S9-mix) | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 15 | 23 | 14 | 17.33 | 4.93 | _ |
Positive control solvent | 20 µL | 19 | 16 | 16 | 17.00 | 1.73 | _ |
Positive control : 2-Nitrofluorene | 2 µL |
423 |
423 |
441 |
429.00 |
10.39 |
17.04 |
Vehicle | 50µL | 14 | 17 | 12 | 14.33 | 2.52 | _ |
| 5000 µg | 22 | 6 | 13 | 13.67 | 8.02 | 0.95 |
Solution of | 1500 µg | 16 | 24 | 17 | 19.00 | 4.36 | 1.33 |
T-A10 BATCH: M22154C | 500 µg | 15 | 18 | 16 | 16.33 | 1.53 | 1.14 |
| 150 µg | 17 | 18 | 18 | 17.67 | 0.58 | 1.23 |
| 50 µg | 16 | 24 | 21 | 20.33 | 4.04 | 1.42 |
TA98 Assay n°2 – with metabolic activation (10 % S9-mix) – with pre-incubation | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 31 | 29 | 130 | 30.00 | 1.00 | _ |
Positive control solvent | 10 µL | 24 | 31 | 27 | 27.33 | 3.51 | _ |
Positive control : 2-Anthramine | 1 µg
|
215 |
218 |
301 |
244.67 |
48.81 |
8.95 |
Vehicle | 50µL | 29 | 23 | 22 | 24.67 | 3.79 | _ |
| 5000 µg | 16 | 27 | 15 | 19.33 | 6.66 | 0.78 |
Solution of | 1500 µg | 33 | 26 | 28 | 29.00 | 3.61 | 1.18 |
T-A10 BATCH: M22154C | 500 µg | 35 | 28 | 21 | 28.00 | 7.00 | 1.14 |
| 150 µg | 31 | 34 | 32 | 32.33 | 1.53 | 1.31 |
| 50 µg | 127 | 27 | 32 | 28.67 | 2.89 | 1.16 |
TA100 Assay n°1 – without metabolic activation (-S9-mix) | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 68 | 60 | 70 | 66.00 | 5.29 | _ |
Positive control solvent | 20 µL | 69 | 57 | 74 | 66.67 | 8.74 | _ |
Positive control : Sodium azide | 20 µg |
1137 |
996 |
1137 |
1069.00 |
81.41 |
16.35 |
Vehicle | 50µL | 69 | 69 | 81 | 73.00 | 6.93 | _ |
| 5000 µg | 43 | 39 | 33 | 38.33 | 5.03 | 0.53 |
Solution of | 1500 µg | 82 | 76 | 85 | 81.00 | 4.58 | 1.11 |
T-A10 BATCH: M22154C | 500 µg | 66 | 77 | 56 | 66.33 | 10.50 | 0.91 |
| 150 µg | 55 | 62 | 71 | 62.67 | 8.02 | 0.86 |
| 50 µg | 71 | 85 | 636 | 73.00 | 11.14 | 1.00 |
TA100 Assay n°1 – with metabolic activation (10 % S9-mix) – without pre-incubation | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 54 | 54 | 62 | 56.67 | 4.62 | _ |
Positive control solvent | 20 µL | 68 | 69 | 65 | 67.33 | 2.08 | _ |
Positive control : 2-Anthramine | 2 µg |
527 |
557 |
531 |
538.33 |
16.29 |
8.00 |
Vehicle | 50µL | 54 | 57 | 66 | 59.00 | 6.24 | _ |
| 5000 µg | 35 | 45 | 33 | 37.67 | 6.43 | 0.64 |
Solution of | 1500 µg | 82 | 67 | 53 | 67.33 | 14.50 | 1.14 |
T-A10 BATCH: M22154C | 500 µg | 74 | 82 | 67 | 74.33 | 7.51 | 1.26 |
| 150 µg | 99 | 69 | 66 | 78.00 | 18.25 | 1.32 |
| 50 µg | 72 | 59 | 82 | 71.00 | 11.53 | 1.20 |
TA100 Assay n°2 – without metabolic activation (-S9-mix) | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 68 | 87 | 66 | 73.67 | 11.59 | _ |
Positive control solvent | 20 µL | 68 | 84 | 57 | 69.67 | 13.58 | _ |
Positive control : Sodium azide | 20 µg
|
1188 |
1124 |
1115 |
1142.33 |
39.80 |
16.40 |
Vehicle | 50µL | 80 | 69 | 46 | 65.00 | 17.35 | _ |
| 5000 µg | 49 | 32 | 29 | 36.67 | 10.79 | 0.56 |
Solution of | 1500 µg | 56 | 60 | 62 | 59.33 | 3.06 | 0.91 |
T-A10 BATCH: M22154C | 500 µg | 61 | 52 | 62 | 58.33 | 5.51 | 0.90 |
| 150 µg | 69 | 72 | 60 | 67.00 | 6.24 | 1.03 |
| 50 µg | 59 | 70 | 57 | 62.00 | 7.00 | 0.95 |
TA100 Assay n°2 – with metabolic activation (10 % S9-mix) – with pre-incubation | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 57 | 66 | 60 | 61.00 | 4.58 | _ |
Positive control solvent | 10 µL | 67 | 71 | 65 | 67.67 | 3.06 | _ |
Positive control : 2-Anthramine | 1 µg |
509 |
498 |
456 |
487.67 |
27.97 |
7.21 |
Vehicle | 50µL | 69 | 61 | 72 | 67.33 | 5.96 | _ |
| 5000 µg | 32 | 47 | 49 | 42.67 | 9.29 | 0.63 |
Solution of | 1500 µg | 74 | 61 | 71 | 68.67 | 6.81 | 1.02 |
T-A10 BATCH: M22154C | 500 µg | 74 | 82 | 71 | 75.67 | 5.69 | 1.12 |
| 150 µg | 69 | 59 | 82 | 70.00 | 11.53 | 1.04 |
| 50 µg | 82 | 79 | 71 | 77.33 | 5.69 | 1.15 |
E. COLI Assay n°1 – without metabolic activation (-S9-mix) | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 50 | 49 | 56 | 51.67 | 3.79 | _ |
Positive control solvent | 10 µL | 61 | 49 | 55 | 55.00 | 6.00 | _ |
Positive control : MMS | 100 µg
|
524 |
531 |
601 |
552.00 |
42.58 |
10.04 |
Vehicle | 50µL | 47 | 66 | 69 | 60.67 | 11.93 | _ |
| 5000 µg | 42 | 38 | 35 | 38.33 | 3.51 | 0.63 |
Solution of | 1500 µg | 55 | 62 | 63 | 60.00 | 4.36 | 0.99 |
T-A10 BATCH: M22154C | 500 µg | 51 | 49 | 48 | 49.33 | 1.53 | 0.81 |
| 150 µg | 49 | 56 | 55 | 53.33 | 3.79 | 0.88 |
| 50 µg | 51 | 49 | 52 | 50.67 | 1.53 | 0.84 |
E. COLI Assay n°1 – with metabolic activation (10 % S9-mix) – without pre-incubation | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 70 | 80 | 82 | 77.33 | 6.43 | _ |
Positive control solvent | 50 µL | 83 | 83 | 90 | 85.33 | 4.04 | _ |
Positive control : 2-Anthramine | 50 µg |
1034 |
1088 |
1003 |
1041.67 |
43.02 |
12.21 |
Vehicle | 50µL | 85 | 79 | 91 | 85.00 | 6.00 | _ |
| 5000 µg | 59 | 58 | 56 | 57.67 | 1.53 | 0.68 |
Solution of | 1500 µg | 79 | 84 | 82 | 81.67 | 2.52 | 0.96 |
T-A10 BATCH: M22154C | 500 µg | 81 | 80 | 90 | 83.67 | 5.51 | 0.98 |
| 150 µg | 89 | 87 | 79 | 85.00 | 5.29 | 1.00 |
| 50 µg | 83 | 85 | 81 | 83.00 | 2.00 | 0.98 |
E. COLI Assay n°2 – without metabolic activation (-S9-mix) | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 47 | 45 | 43 | 45.00 | 2.00 | _ |
Positive control solvent | 10 µL | 54 | 63 | 43 | 53.33 | 10.02 | _ |
Positive control : MMS | 100 µg
|
445 |
535 |
374 |
451.33 |
80.69 |
8.46 |
Vehicle | 50µL | 51 | 48 | 65 | 54.67 | 9.07 | _ |
| 5000 µg | 38 | 35 | 32 | 35.00 | 3.00 | 0.64 |
Solution of | 1500 µg | 45 | 56 | 60 | 53.67 | 7.77 | 0.98 |
T-A10 BATCH: M22154C | 500 µg | 54 | 62 | 61 | 59.00 | 4.36 | 1.08 |
| 150 µg | 59 | 46 | 45 | 50.00 | 7.81 | 0.91 |
| 50 µg | 43 | 63 | 44 | 50.00 | 11.27 | 0.91 |
E. COLI Assay n°2 – with metabolic activation (10 % S9-mix) – with pre-incubation | |||||||
Serie |
Dose/Plate | Plate |
Mean |
Standard deviation |
R | ||
n° 1 | n° 2 | n° 3 | |||||
Negative control | 100 µL | 70 | 69 | 76 | 71.67 | 6,24 | _ |
Positive control solvent | 12,5 µL | 80 | 100 | 63 | 81.00 | 12,06 | _ |
Positive control : Dimethylbenzanthracene | 12.5 µg
|
1118 |
894 |
772 |
4928.00 |
27,57 |
11.16 |
Vehicle | 50µL | 81 | 72 | 86 | 79.67 | 7,02 | _ |
| 5000 µg | 45 | 44 | 39 | 42.67 | 3,06 | 0.54 |
Solution of | 1500 µg | 75 | 83 | 82 | 80.00 | 6,03 | 1.00 |
T-A10 BATCH: l19319E | 500 µg | 81 | 77 | 76 | 78.00 | 3,61 | 0.98 |
| 150 µg | 80 | 76 | 72 | 76.00 | 9,64 | 0.95 |
| 50 µg | 80 | 83 | 68 | 77.00 | 13,75 | 0.97 |
Applicant's summary and conclusion
- Conclusions:
- The test item did not induce any mutagenic change in the bacterial reverse mutation test in any of the strains tested with and without metabolic activation up to 5000 μg/plate.
- Executive summary:
A bacterial reverse mutation test was conducted on the test substance according to OECD guideline 471 under GLP conditions. Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 and Escherichia coli WP2 uvrA were exposed to concentrations of the test substance ranging from 50 to 5000 μg/plate in DMSO, with and without metabolic activation, based on preliminary solubility and cytotoxicity tests. The metabolic activation system (S9 fraction) was prepared from Sprague Dawley rat liver homogenate. Two independent assays were performed in all strains: an initial mutation test (plate incorporation method) and a confirmatory mutation test (plate incorporation method without S9
and pre-incubation method with S9). Untreated, solvent controls and strain specific positive controls were included in the assays and the values obtained were within ranges of the historical control values of the laboratory in all strains. All validity criteria were fulfilled. The test item did not induce any significant increase in the number of revertants in any of the strains tested, with and without metabolic activation, up to 5000 μg/plate. Based on these results, the test item can be considered as not mutagenic according to the OECD TG 471.
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