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EC number: 258-296-4 | CAS number: 53018-24-9
- 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
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- 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
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- Endpoint summary
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- Environmental data
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- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Under the conditions of this study, the test material was not mutagenic to the bacterial strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100 in the presence or absence of metabolic activation.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 19 October 1985 to 04 November 1985
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- The purpose of this mutagenicity test was to investigate the potential of the test material to induce reverse mutations in histidine dependent strains of Salmonella typhimurium. This was done by incubating the bacteria with the test material on selective media (histidine deficient) which inhibits the growth of histidine dependent bacteria, but permits the growth of histidine independent revertants. Since some chemicals are themselves nonmutagenic but are metabolised to mutagenic metabolites the test was conducted both with and without liver preparation (S-9 mix) which has the function of metabolising the test material. A preliminary toxicity test was required in order to establish dose levels - the test material was tested up to the maximum concentration permitted by toxicity up to a maximum of 5000 µg/plate for non-toxic materials. At least five dose levels were tested together with positive and negative controls.
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- Histidine dependent strains of Salmonella typhimurium.
- Species / strain / cell type:
- S. typhimurium, other: TA1537, TA1535, TA100, TA1538 & TA98
- Details on mammalian cell type (if applicable):
- - The strains carry the uvrB mutation and are therefore repair deficient. This results in an increased sensitivity in detecting mutagens.
- The strains also carry the deep rough mutation (rfa) which causes a partial loss in the polysaccharide coat of the bacteria and therefore allows penetration of large molecules.
- Strains TA98 and TA100 contain the R factor plasmid pKM101 and are more sensitive than the parent strains TA1538 and TA1535 respectively which do not contain the plasmid. The plasmid containing strains are Ampicillin resistant.
- The strains are routinely checked with diagnostic mutagens for strain specificity, for UV sensitivity, for sensitivity to Crystal Violet, and for Ampicillin resistance if appropriate (Ames et al, 1975). - Metabolic activation:
- with and without
- Metabolic activation system:
- S-9 mix
- Test concentrations with justification for top dose:
- - Dose range-finding test: 5000, 500, 50 and 5 μg/plate.
- Main test: 5000, 1500, 500, 150 and 50 μg/plate.
The top dose level was chosen after an evaluation of the results of the preliminary toxicity test. It was either the lowest dose showing toxicity in the preliminary test or the next dose down on a half log10 scale, according to the judgement of the Experimental Supervisor. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- other: 2-aminoanthracene
- Details on test system and experimental conditions:
- PRELIMINARY TEST
The preliminary toxicity test followed the same test method as the main test (see below), with the exception that four concentrations of test material and duplicate plates were used. The concentrations for the toxicity test were 5000, 500, 50 and 5 μg/plate.
MAIN TEST
- Sub-cultures of bacteria were grown in nutrient broth at 37°C for 16 hrs to provide approximately 2 x 10^9 bacteria per mL. Two mL aliquots of molten soft agar containing 0.5 mM of histidine HCl, and 0.5mM biotin were dispensed into glass tubes and maintained at 45°C. To triplicate plates 0.5 mL of S-9 mix (with metabolism) or 0.5mL of 0.1 M phosphate buffer, pH 7.4 (without metabolism), 0.1 mL of bacterial suspension (2 x 10^9 bacteria/mL) and 0.1 mL of test material in DMSO or other appropriate solvent at each of five concentrations were added.
- The contents of the tubes were thoroughly mixed and poured onto petri dishes containing 30 mL of minimal agar. The plates were incubated at 37°C for 72 hours.
- The number of colonies on each plate was counted and the mean number of revertant colonies per treatment group was calculated.
- The sterility of the S-9 mix and test solutions was confirmed by the incubation of aliquots of the liquids on nutrient agar.
- For the pre-incubation method, 0.5 mL of S-9 mix or 0.5 mL of 0.1 M phosphate buffer, pH 7.4, was incubated for 30 minutes at 37°C with 0.1 mL of bacterial culture and 0.01 mL of test material solution. After incubation, two mL of melted (45°C) soft agar containing 0.5 mM histidine HCl and 0.5 mM biotin was added, mixed, and overlaid onto minimal agar plates. The plates were incubated and colonies counted as above. - Evaluation criteria:
- For each strain the mean number of revertants at each treatment group both with and without metabolism was compared with those of the negative control groups. The test material was considered to be a positive mutagen if a dose related increase in revertants, up to at least a doubling of the control values, was obtained in two independent tests.
- Key result
- Species / strain:
- S. typhimurium, other: TA1535, TA1537, TA1538, TA98 & TA100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- - The test material was toxic towards the tester strains.
- The test material did not induce significant number of revertant colonies with any of the five strains either in the presence or absence of S-9 mix.
- It was concluded that no evidence of mutagenic potential of the test material was obtained in the bacterial test system at the concentrations used. However, toxicity was observed at 1500 μg/plate towards all strains without activation. In assays with activation, the toxicity was observed at 1500 µg/plate towards TA 100 and at 5000 µg/plate towards all strains. - Conclusions:
- Under the conditions of this study, the test material was not mutagenic to the bacterial strains in the presence or absence of metabolic activation.
- Executive summary:
The genetic toxicity of the test material was investigated in a bacterial reverse mutation assay with Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100.
The test material was formulated in DMSO and tested on the bacteria at dose levels of: 5000, 1500, 500, 150 and 50 μg/plate.
The dose levels were selected following a preliminary dose range-finding test. DMSO was used as the solvent control and appropriate positive controls were included to show the suitability of the test system. The test material was examined in the presence and absence of metabolic activation in the form of S-9 mix.
Under the condition of the study the test material was toxic towards all of the tester strains. The test material did not induce significant number of revertant colonies with any of the five strains either in the presence or absence of S-9 mix. It was concluded that no evidence of mutagenic potential of the test material was obtained in the bacterial test system at the concentrations used. However, toxicity was observed at 1500 μg/plate towards all strains without activation. In assays with activation, the toxicity was observed at 1500 µg/plate towards TA 100 and at 5000 µg/plate towards all strains.
The test material was therefore considered not mutagenic to the bacterial strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100 in the presence or absence of metabolic activation.
Reference
Table 1: Summary of First Main Experiment
± S9 Mix |
Concentration (µg/plate) |
Mean number of colonies/plate |
||||
TA100 |
TA1535 |
TA1538 |
TA98 |
TA1537 |
||
- |
Solvent 50 150 500 1500 5000 |
208 205 202 176 - - |
47 42 44 44 - - |
17 17 16 16 - - |
49 50 50 41 - - |
17 19 17 17 - - |
+ |
Solvent 50 150 500 1500 5000 |
154 148 150 134 - - |
27 27 24 23 19 - |
22 23 23 22 18 - |
32 32 29 30 25 - |
19 19 18 18 12 - |
Positive Controls |
||||||
- |
Name |
SA |
SA |
2NF |
2NF |
9AA |
Concentration (µg/plate) |
5 |
5 |
10 |
10 |
20 |
|
Mean no. colonies/plate |
>1000 |
>1000 |
>1000 |
>1000 |
123 |
|
+ |
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
Concentration (µg/plate) |
2 |
2 |
2 |
2 |
2 |
|
Mean no. colonies/plate |
825 |
96 |
390 |
489 |
134 |
9AA = 9-aminoacridine
2AA = 2-aminoanthracene
SA = Sodium azide
2NF = 2-Nitrofluorene
Table 2: Summary of Repeat Main Experiment
± S9 Mix |
Concentration (µg/plate) |
Mean number of colonies/plate |
||||
TA100 |
TA1535 |
TA1538 |
TA98 |
TA1537 |
||
- |
Solvent 50 150 500 1500 5000 |
177 177 168 156 - - |
49 47 49 41 - - |
20 18 23 19 - - |
56 62 64 60 - - |
18 24 20 17 - - |
+ |
Solvent 50 150 500 1500 5000 |
145 146 147 134 - - |
28 27 22 20 14 - |
28 25 29 23 19 - |
34 32 34 31 23 - |
18 16 19 18 13 - |
Positive Controls |
||||||
- |
Name |
SA |
SA |
2NF |
2NF |
9AA |
Concentration (µg/plate) |
5 |
5 |
10 |
10 |
20 |
|
Mean no. colonies/plate |
>1000 |
>1000 |
>1000 |
>1000 |
132 |
|
+ |
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
Concentration (µg/plate) |
2 |
2 |
2 |
2 |
2 |
|
Mean no. colonies/plate |
755 |
92 |
430 |
444 |
106 |
9AA = 9-aminoacridine
2AA = 2-aminoanthracene
SA = sodium azide
2NF = 2 -nitrofluorene
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
The genetic toxicity of the test material was investigated in a bacterial reverse mutation assay with Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).
The test material was formulated in DMSO and tested on the bacteria at dose levels of: 5000, 1500, 500, 150 and 50 μg/plate.
The dose levels were selected following a preliminary dose range-finding test. DMSO was used as the solvent control and appropriate positive controls were included to show the suitability of the test system. The test material was examined in the presence and absence of metabolic activation in the form of S-9 mix.
Under the condition of the study the test material was toxic towards all of the tester strains. The test material did not induce significant number of revertant colonies with any of the five strains either in the presence or absence of S-9 mix. It was concluded that no evidence of mutagenic potential of the test material was obtained in the bacterial test system at the concentrations used. However, toxicity was observed at 1500 μg/plate towards all strains without activation. In assays with activation, the toxicity was observed at 1500 µg/plate towards TA 100 and at 5000 µg/plate towards all strains.
The test material was therefore considered not mutagenic to the bacterial strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100 in the presence or absence of metabolic activation.
Justification for classification or non-classification
In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance does not require classification with respect to genetic toxicity.
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