Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 200-625-0 | CAS number: 66-27-3
- 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 mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Justification for type of information:
- Data is from peer reviewed publication
Data source
Reference
- Reference Type:
- publication
- Title:
- Different Types of Combination Effects for the Induction of Micronuclei in Mouse Lymphoma Cells by Binary Mixtures of the Genotoxic Agents MMS, MNU, and Genistein
- Author:
- Werner K. Lutz, Oliver Tiedge, Roman W. Lutz, and Helga Stopper
- Year:
- 2 005
- Bibliographic source:
- TOXICOLOGICAL SCIENCES 86(2), 318–323 (2005)
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- other: Refer below principle
- Principles of method if other than guideline:
- In vitro induction of micronuclei was tested using L5178Y mouse lymphoma cells by Methyl methanesulfonate
- GLP compliance:
- not specified
- Type of assay:
- mammalian cell gene mutation assay
Test material
- Reference substance name:
- Methyl methanesulphonate
- EC Number:
- 200-625-0
- EC Name:
- Methyl methanesulphonate
- Cas Number:
- 66-27-3
- Molecular formula:
- C2H6O3S
- IUPAC Name:
- methyl methanesulfonate
- Details on test material:
- - Name of test material: Methyl methanesulfonate (MMS)
- Molecular formula: C2H6O3S
- Molecular weight: 110.1324 g/mol
- Substance type: Organic
- Physical state: Liquid
- Purity: No data available
- Impurities (identity and concentrations): No data available
Constituent 1
- Specific details on test material used for the study:
- - Name of test material: Methyl methanesulfonate (MMS)
- Molecular formula: C2H6O3S
- Molecular weight: 110.1324 g/mole
- Substance type: Organic
- Physical state: Liquid
- Purity: No data available
- Impurities (identity and concentrations): No data available
Method
- Target gene:
- No data
Species / strain
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPMI 1640 cell culture medium supplemented with antibiotics,
0.25 mg L-glutamine/ml, 107 µg sodium pyruvate /ml, and 10% heat inactivated horse serum
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: No data available
- Periodically checked for karyotype stability: No data available
- Periodically "cleansed" against high spontaneous background: No data available - Additional strain / cell type characteristics:
- not specified
- Cytokinesis block (if used):
- No data
- Metabolic activation:
- not specified
- Metabolic activation system:
- No data
- Test concentrations with justification for top dose:
- 0, 100, 200, 300 µM
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The chemical is soluble in DMSO
Controls
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- not specified
- Positive control substance:
- not specified
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
- Cell density at seeding (if applicable): 200000 cells per mL
DURATION
- Preincubation period: No data available
- Exposure duration: 4 hrs
- Expression time (cells in growth medium): 20 hrs
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): 24 hrs
SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): Cytochalasin B
STAIN (for cytogenetic assays): Acridine orange
NUMBER OF REPLICATIONS: Duplicate
NUMBER OF CELLS EVALUATED: Two thousand cells (1000 per slide) were evaluated for each treatment. The score (MN) was the number of cells containing one or more micronuclei per 1000 binucleated cells (BN cells). The percentage of binucleated cells was evaluated as a marker of cell proliferation.
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available
OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available
OTHER: No data available - Rationale for test conditions:
- No data available
- Evaluation criteria:
- The percentage of binucleated cells was evaluated as a marker of cell proliferation.
- Statistics:
- The hypothesis of response addition (addition of net effects of the components) was tested by the sign test. Under the null hypothesis of response addition and the assumption of symmetrical errors, the observed number of cells with micronuclei is with equal probability greater or less than the number of cells with micronuclei calculated for response addition.
The hypothesis of dose addition was tested with a linear model, using the logarithm of the score MN to linearize the sublinear dose responses for the components in a parallel manner. For the test of deviation from dose addition,
Testing for interaction according to
Log10 (MN) = a +(bXA)+(c X B) +(d X A X B)+Ɛ (error term)
Letters A and B stand for the concentrations of the two chemicals. AX B is the interaction term. The coefficients a, b, c, and d were estimated by linear regression. The error was proportional to the score MN. For the statistical handling of the replicates, random effects for the parameters were added if this resulted in a model improvement (mixed effects model).
Note: The absence of an interaction (d ¼ 0) is equivalent to dose addition, as can be shown by replacing B for A by normalization with c/b (last line):
MN= 10 (a+b X A+c X B+c) = 10a X 10(bXA) X 10(c X B) X 10c
=10a X 10(bXA) X 10(b X c/b X B) X 10c
= 10a X (10b)A X 10(c/b X B) X 10c
=10a X (10b)(A+c/bXB) X 10c
Results and discussion
Test results
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- not specified
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Additional information on results:
- No data available
Applicant's summary and conclusion
- Conclusions:
- Methyl methanesulfonate (MMS) tested at a concentration of 0, 100, 200, 300 µM showed the induction of micronuclei in the mouse Lymphoma L5178Y cells and hence is positive for gene mutation in vitro.
- Executive summary:
Analysis for the induction of micronuclei in L5178Y mouse lymphoma cells by the methylating agent methyl methanesulfonate (MMS) was performed. The test chemical was used at dose levels of 0, 100, 200 and 300 µM. Pilot experiment was performed to determine the tange for the test and the concentration range used was defined by similar effect magnitude, in order not to exceed the response range of the assay that may be limited by cytotoxicity. Mouse lymphoma L5178Y cells, clone 3.7.2c were cultured in suspension in RPMI 1640 cell culture medium supplemented with antibiotics, 0.25 mg L-glutamine/ml, 107 µg sodium pyruvate/ml, and 10% heat inactivated horse serum. Methyl methanesulfonate (MMS) was dissolved in DMSO and added to L5187Y mouse lymphoma cells at a density of 2 X 105cells per ml. The cells were washed twice after 4 hrs chemical exposure and cytochalasin B was added. Cytochalasin B remained in the culture for the entire expression time of 20 h until the cells were harvested. Cytospin preparations on glass slides were prepared. After 2 h in methanol at -20C, the slides were incubated with acridine orange for 5 min, washed twice with Sorensen buffer for 5 min, and mounted for microscopy. Two thousand cells (1000 per slide) were evaluated for each treatment. The score (MN) was the number of cells containing one or more micronuclei per 1000 binucleated cells (BN cells). The percentage of binucleated cells was evaluated as a marker of cell proliferation. components. Two completely independent sets of experiments were run for each pairwise combination. In addition, controls were run in duplicate. A dose response relationship was observed for micronucleus induction in L5178Y mouse lymphoma cells. Methyl methanesulfonate (MMS) tested at a concentration of 0, 100, 200, 300 µM showed the induction of micronuclei in the mouse Lymphoma L5178Y cells and hence is positive for gene mutation in vitro.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.