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EC number: 291-367-8 | CAS number: 90387-90-9
- 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
- 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
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 29 October 2010 - 16 March 2011
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Study was generated according to internationally accepted guidelines.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 011
- Report date:
- 2011
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- GLP according to Chemikaliengesetz and Directive 2004/9/EC
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Graphite, acid-treated
- EC Number:
- 291-367-8
- EC Name:
- Graphite, acid-treated
- Cas Number:
- 90387-90-9
- Molecular formula:
- C(Carbon) + acid + oxidant Not available by EC, (CnX mHX)
- IUPAC Name:
- carbon
- Details on test material:
- - Name of test material (as cited in study report): graphite, acid treated
- Substance type: inorganic
- Physical state: soild
- Lot/batch No.: 1706
- Expiration date of the lot/batch: not applicable
- Storage condition of test material: at room temperature
- Colour: grey
- Density: 2.2 g/cm³
- Particle size range: 80% < 50 mesh
Constituent 1
Method
- Target gene:
- Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 102.
Species / strain
- Species / strain / cell type:
- S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537, TA 102
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- other: TA 98: his D 3052; rfa-; uvrB-; R-factor: frame shift mutations TA 100: his G 46; rfa-; uvrB-; R-factor: base-pair substitutions TA 1535: his G 46; rfa-; uvrB-: base-pair substitutions TA 1537: his C 3076; rfa-; uvrB-:
- Metabolic activation:
- with and without
- Metabolic activation system:
- 2-AA; 2-aminoanthracene
- Test concentrations with justification for top dose:
- The following concentrations of the test item extracts were prepared and used in the experiments:
10, 20, 40, 60, 80 and 100 %.
The 100 % extract concentration corresponds to a weight/volume-ratio of 0.2 g/mL 0.9 % NaCl resp. DMSO. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: 0.9% NaCl-solution (as polar extraction medium); DMSO (non-polar extraction medium).
- Weight/volume ratio: 0.2 g/mL according to ISO 10993-3, 2003 and ISO 10993-12, 2007
- Justification for choice of solvent/vehicle: stability of test substance in polar and non-polar extraction medium
Controlsopen allclose all
- Untreated negative controls:
- yes
- Remarks:
- extraction medium and A. dest
- Positive controls:
- yes
- Remarks:
- S. typhimurium: TA 100, TA 1535; without metabolic activation
- Positive control substance:
- sodium azide
- Remarks:
- Dissolved in: Aqua dest. Concentration: 10 µg /plate
- Positive controls:
- yes
- Remarks:
- S. typhimurium: TA 98, TA 1537; without metabolic activation
- Positive control substance:
- other: 4-NOPD; 4-nitro-o-phenylene-diamine
- Remarks:
- Dissolved in: DMSO Concentrations: 10 µg/plate for TA 98 40 µg/plate for TA 1537
- Positive controls:
- yes
- Remarks:
- S. typhimurium: TA 102; without metabolic activation
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- Dissolved in: Aqua dest. Concentration: 1 µL/plate
- Positive controls:
- yes
- Remarks:
- S. typhimurium: TA 98, TA 100, TA 1535, TA 1537 and TA 102; with metabolic activation
- Positive control substance:
- other: 2-AA; 2-aminoanthracene
- Remarks:
- Dissolved in: DMSO Concentrations: 2.5 µg/plate ; 10 µg/plate for TA 102
- Details on test system and experimental conditions:
- PREPARATION OF THE TEST ITEM:
The test item was extracted in a polar extraction medium [0.9% NaCl Lot No.: Delta Select / 251002T-1 (physiological solution)] and in a non-polar extraction medium (DMSO Lot No.: Applichem / 9U009953) for 72 (+/- 2) h at 37 (+/- 1) °C at a weight/volume ratio of 0.2 g/mL according to ISO 10993-3, 2003 and ISO 10993-12, 2007. The extraction procedure did not reveal any abnormalities in the extraction medium or the test item. After extraction with the non-polar extraction medium DMSO a greyish colouring of the extract was observed. After extraction the polar and non-polar extracts were centrifuged at room temperature for 5 minutes at 1000 g. The polar extract was further processed by sterile filtration (filter manufacturer: Whatman, batch: D102229, material: cellulose acetate, pore size: 0.2 μm).
TEST SYSTEM
BACTERIA:
Five strains of S. typhimurium with the following characteristics were used:
TA 98:
his D 3052; rfa-; uvrB-; R-factor: frame shift mutations
TA 100:
his G 46; rfa-; uvrB-; R-factor: base-pair substitutions
TA 1535:
his G 46; rfa-; uvrB-: base-pair substitutions
TA 1537:
his C 3076; rfa-; uvrB-: frame shift mutations
TA 102:
his G 428 (pAQ1); rfa-; R-factor: base-pair substitutions
PREPARATION OF BACTERIA:
Samples of each tester strain were grown by culturing for 12 h at 38.5 °C in nutrient broth to the late exponential or early stationary phase of growth (approx. 109 cells/mL). The nutrient medium consists per litre:
8 g Nutrient Broth
5 g NaCl
A solution of ampicillin (125 µL, 10 mg/mL) (TA 98, TA 100 and TA 102) was added in order to retain the phenotypic characteristics of the strain.
AGAR PLATES:
The Vogel-Bonner Medium E agar plates with 2 % glucose used in the Ames Test were prepared by BSL BIOSERVICE GmbH or provided by an appropriate supplier. Quality controls were performed.
Vogel-Bonner-salts contain per litre:
10 g MgSO4 x 7 H2O
100 g citric acid
175 g NaNH4HPO4 x 4 H2O
500 g K2HPO4
Sterilisation was performed at 121 °C in an autoclave.
Vogel-Bonner Medium E agar plates contain per litre:
15 g Agar Agar
20 mL Vogel-Bonner salts
50 mL glucose-solvent (40%)
Sterilisation was performed at 121 °C in an autoclave.
OVERLAY AGAR:
The overlay agar contains per litre:
7.0 g Agar Agar
6.0 g NaCl
10.5 mg L-histidine x HCl x H2O
12.2 mg biotin
Sterilisation was performed at 121 °C in an autoclave.
S9 HOMOGENATE:
The S9 liver microsomal fraction was prepared at BSL BIOSERVICE GmbH. Male Wistar rats were induced with phenobarbital (80 mg/kg bw) and β-naphthoflavone (100 mg/kg bw) for three consecutive days by oral route.
The following quality control determinations are performed:
a) Biological activity in:
- the Salmonella typhimurium assay using 2-aminoanthracene
- the mouse lymphoma assay using benzo[a]pyrene
- the chromosome aberration assay using cyclophosphamide.
b) Sterility Test:
A stock of the supernatant containing the microsomes was frozen in aliquots of 2 and 4.5 mL and stored at <= -75 °C.
The protein concentration in the S9 preparation (Lot: 230910) was 36 mg/mL. The S9 mix preparation was performed according to Ames et al.
PREPARATION OF S9 MIX:
The S9 mix preparation was performed according to Ames et al. 100 mM of ice cold sodium-ortho-phosphate-buffer, pH 7.4, was added to the following pre-weighed sterilised reagents to give final concentrations in the S9 mix of:
8 mM MgCl2
33 mM KCl
5 mM glucose-6-phosphate
4 mM NADP
This solution was mixed with the liver 9000 x g supernatant fluid in the following proportion:
co-factor solution 9.5 parts
liver preparation 0.5 parts
During the experiment the S9 mix was stored on ice.
EXPOSURE CONCENTRATIONS:
The test item was tested with the following extract concentrations in the experiments:
10, 20, 40, 60, 80 and 100% - Evaluation criteria:
- CRITERIA OF VALIDITY:
A test is considered acceptable if for each strain:
- the bacteria demonstrate their typical responses to ampicillin (TA 98, TA 100, TA 102)
- the control plates with and without S9 mix are within the following ranges (mean values of the spontaneous reversion frequency are within the historical control data range):
-S9 +S9
-------------------------
TA 98: 18 - 46 18 - 57
TA 100: 77 - 163 78 - 165
TA 1535: 5 - 29 5- 27
TA 1537: 5 - 30 5 - 36
TA 102: 164 - 390 163 - 472
- corresponding background growth on negative control, extract medium control and test plates is observed.
- the positive controls show a distinct enhancement of revertant rates over the control plate.
EVALUATION OF MUTAGENICITY:
The Mutation Factor is calculated by dividing the mean value of the revertant counts through the mean values of the extract medium control (the exact and not the rounded values are used for calculation).
A test item is considered as mutagenic if:
- a clear and dose-related increase in the number of revertants occurs and/or
- a biologically relevant positive response for at least one of the dose groups occurs
in at least one tester strain with or without metabolic activation.
A biologically relevant increase is described as follows:
- if in tester strains TA 98, TA 100 and TA 102 the number of reversions is at least twice as high
- if in tester strains TA 1535 and TA 1537 the number of reversions is at least three times higher
than the reversion rate of the extract medium control.
A test item producing neither a dose related increase in the number of revertants nor a reproducible biologically relevant positive response at any of the dose groups is considered to be non-mutagenic in this system. - Statistics:
- According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary.
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 102
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
Any other information on results incl. tables
Extracts (polar and non-polar) of graphite, acid treated were investigated for their potential to induce gene mutations according to the pre-incubation method using Salmonella typhimurium tester strains TA 98, TA 100, TA 1535, TA 1537 and TA 102.
Several concentrations of the test item extracts were used. Each assay was conducted with and without metabolic activation. The concentrations, including the controls, were tested in triplicate. The following concentrations of the test item extracts were prepared and used in the experiments:
10, 20, 40, 60, 80 and 100%
The 100% extract concentration corresponds to a weight/volume-ratio of 0.2 g/mL 0.9% NaCl resp. DMSO.
No toxic effects of the test item extracts were observed in any of the five tester strains used up to the highest extract concentrations evaluated (with and without metabolic activation) in experiment I and II. The reduction in the number of revertants down to a mutation factor of 0.4 found in tester strain TA 1535 at an extract concentration of 40% (with metabolic activation) and down to a mutation factor of 0.5 found in tester strain TA 1537 at an extract concentration of 60% (with metabolic activation), both found in experiment II, was regarded as not biologically relevant due to lack of a dose-response relationship.
No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed following treatment with extracts (polar and non-polar) of graphite, acid treated at any concentration level, neither in the presence nor absence of metabolic activation in both experiments.
The reference mutagens induced a distinct increase of revertant colonies indicating the validity of the experiments.
A more comprehensive overview of the test results can be seen from the document attached below.
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative
In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item extracts did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used.
Therefore, extracts (polar and non-polar) of graphite, acid treated are considered to be non-mutagenic in this bacterial reverse mutation assay. - Executive summary:
In order to investigate the potential of extracts (polar and non-polar) of graphite, acid treated for their ability to induce gene mutations the pre-incubation test was performed with theS almonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 102.
The test item extracts were tested at several concentrations. The assays were conducted with and without metabolic activation. The concentrations, including the controls, were tested in triplicate. The following concentrations of the test item extracts were prepared and used in the experiments:
10, 20, 40, 60, 80 and 100%
The 100% extract concentration corresponds to a weight/volume-ratio of 0.2 g/mL 0.9 % NaCl resp. DMSO.
No toxic effects of the test item extracts were noted in any of the five tester strains used up to the highest extract concentrations evaluated (with and without metabolic activation) in experiment I and II.
No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed following treatment with extracts (polar and non-polar) of graphite, acid treated at any concentration level, neither in the presence nor absence of metabolic activation in experiment I and II.
The reference mutagens induced a distinct increase of revertant colonies indicating the validity of the experiments.
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