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EC number: 206-126-4 | CAS number: 302-72-7
- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
in vitro gene mutation study in bacteria, OECD 471 (Ames test), strains TA 1535, TA 1537, TA 92, TA 98, TA94 and TA 100 of S. typhimurium, max. dose 20 mg/plate, with metbolic activation (S9 -Mix), Result: negative
in vitro cytogenicity / chromosome aberration study in mammalian cells, OECD 473, chromosomal aberration test Chinese hamster lung fibroblast CHO cells, max. dose 1 mg/plate, without metabolic activation, Result: negative
Link to relevant study records
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- GLP compliance:
- not specified
- Type of assay:
- other: in vitro mammalian chromosome aberration test (migrated information)
- Species / strain / cell type:
- other: Chinese hamster fibroblast CHL
- Details on mammalian cell type (if applicable):
- CELLS USED
- Source of cells: The cell line was originally established from the lung of a newborn female at the Cancer Research Institute, Tokyo (Koyama, Utakoji & Ono, 1970)
- Methods for maintenance in cell culture if applicable: maintained by 4-day passages in Medium (see below)
- Modal number of chromosomes: 25
- Normal (negative control) cell cycle time: 15hr
MEDIA USED
- Type and identity of media including CO2 concentration if applicable: MEM (GIBCO) supplemented with 10 % calf serum
- Properly maintained: yes - Metabolic activation:
- without
- Test concentrations with justification for top dose:
- three different doses
max. 1.0 mg/ml - Vehicle / solvent:
- physiological saline
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- no
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: The cells were exposed to each sample at three different doses for 24 and 48 hr. In the present studies, no metabolic activation systems were applied. The maximum dose of each sample was selected by a preliminary test in which the dose needed for 50% cell-growth inhibition was estimated using a cell densitometer (Monocellater, Olympus Co., Ltd). Previous studies indicated that the osmotic pressure of the medium generally rose with sample concentrations of more than 10 mM, so that the maximum dose for some samples was limited to around this level, at which cytotoxic effects were not necessarily observed.
SPINDLE INHIBITOR (cytogenetic assays): Colcemid (final concn 0.2 /µg/ml)
STAIN (for cytogenetic assays): After air-drying, the slides were stained with Giemsa solution (1.5%, at pH 6.8; E. Merck) for 12-15 min.
METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: Chromosome preparations were made as follows. Colcemid (final concn 0.2 /µg/ml) was added to the culture 2 hr before cell harvesting. The cells were then trypsinized and suspended in a hypotonic KCl solution (0.075 M) for 13 min at room temperature. After centrifugation the cells were fixed with acetic acid-methanol (1:3, v/v) and spread on clean glass slides. After air-drying, the slides were stained with Giemsa solution (1.5%, at pH 6.8; E. Merck) for 12-15 min.
NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells): A hundred well-spread metaphases were observed under the microscope (x 600 with a no-cover objective lens).
DETERMINATION OF CYTOTOXICITY
The incidence of polyploid cells as well as of cells with structural chromosomal aberrations such as chromatid or chromosome gaps, breaks, exchanges, ring formations, fragmentations and others, was recorded on each culture plate. Untreated cells and solvent-treated cells served as negative controls, in which the incidence of aberrations was usually less than 3.0%. The results were considered to be negative if the incidence was less than 4.9 %, equivocal if it was between 5.0 and 9.9 %, and positive if it was more than 10. 0%.
- Evaluation criteria:
- The incidence of polyploid cells as well as of cells with structural chromosomal aberrations such as chromatid or chromosome gaps, breaks, exchanges, ring formations, fragmentations and others, was recorded on each culture plate. Untreated cells and solvent-treated cells served as negative controls, in which the incidence of aberrations was usually less than 3.0%. The maximum dose for positive results in the Chromosome aberration test represents the dose at which the maximum effect was obtained, while the maximum dose for negative results represents the highest non-cytotoxic dose used in the experimet.
A result was considered positive if the total incidence of cells with aberrations (including gaps) was 10.0 % or more, equivocal if the incidence was between 5.0 and 9.9 % and negative if the incidence was 4.9 % or less. - Statistics:
- not reported
- Key result
- Species / strain:
- other: Chinese hamster fibroblast cell line CHL
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not examined
- Additional information on results:
- The results of the chromosomal aberration tests:
The maximum dose of test substance used: 1.0 mg/ml
The solvent used: physiological saline
The incidence of polyploid cells at 48 hr: 1.0 %
The incidence of cells with structural aberrations at 48 hr after treatment: 1.0 %
The final result: D,L-Alanine does not induce chromosomal aberrations in CHO cells. - Conclusions:
- DL-Alanine was tested negative for chromosome aberration without metabolic activation.
- Executive summary:
In a chromosomal aberration test Chinese hamster lung fibroblast CHO cells were exposed to DL-Alanine in physiological saline in at least three different concentrations up to 1 mg/plate for 24 and 48 hours without metabolic activation. The results were considered to be negative since the incidence of polyploid cells after 48 hr is 1.0% and the incidence of cells with structural aberrations at 24 or 48 hr after treatment is 1.0% as well.
The study was performed in accordance with OECD guideline 473 with minor deviations. Therefore, the results can be considered to be reliable.
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- The recommended 5th bacterial strain (E. coli WP2 uvrA, or E. coli WP2 uvrA (pKM101), or S. typhimurium TA102) is not used in this study
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his-
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Remarks:
- S.typh. strain TA94 and TA92 also used
- Metabolic activation:
- with
- Metabolic activation system:
- S-9
- Test concentrations with justification for top dose:
- at least five different concentrations, up to 20.0 mg/plate
- Vehicle / solvent:
- Phosphate buffer
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Details on test system and experimental conditions:
- Cells cultured overnight were pre-incubated with both the test sample and the S-9 mix for 20 min at 37 °C before plating. Duplicate plates were used for each of six different concentrations of the sample. The number of revertant (his+) colonies was scored after incubation at 37 °C for 2 days. The result was considered positive if the number of colonies found was twice the number in the control (exposed to the appropriate solvent or untreated). If no reasonable dose response was detected, additional experiments using different doses or induced mutation frequency assays (Yoshikawa, Nakadate, Watabe et al. 1980) were performed.
- Evaluation criteria:
- The result was considered positive if the number of his+ colonies found was twice the number in the control (exposed to the appropriate solvent or untreated). The maximum dose for positive results in the Ames test represents the dose at which the maximum effect was obtained, while the maximum dose for negative results represents the highest non-cytotoxic dose used in the experimet.
A negative result indicates that no significant increases in the numbers of revertant colonies were detected in any S.typhimurium strains at the maximum dose. - Statistics:
- not reported
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not examined
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not examined
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not examined
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not examined
- Key result
- Species / strain:
- S. typhimurium, other: TA92 and TA94
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not examined
- Conclusions:
- DL-Alanine was negative with metabolic activation in all trains tested.
- Executive summary:
In a reverse gene mutation test in bacteria (Ames test), strains TA 1535, TA 1537, TA 92, TA 98, TA94 and TA 100 of S. typhimurium were exposed to DL-Alanine in phosphate buffer in at least six different concentrations up to 20 mg/plate in the presence of mammalian metabolic activation (rat S9, preincubation). There was no evidence of induced mutant colonies over background.
The study was performed in accordance to OECD guideline 471 with minor deviations. Therefore, the results can be considered to be reliable.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
in vitro gene mutation study in bacteria
In a key reverse gene mutation test (Ishidate, 1984: S.typh) in bacteria (Ames test), strains TA 1535, TA 1537, TA 92, TA 98, TA94 and TA 100 of S. typhimurium were exposed to DL-Alanine in phosphate buffer in at least six different concentrations up to 20 mg/plate in the presence of mammalian metabolic activation (rat S9, preincubation). There was no evidence of induced mutant colonies over background. The study was performed in accordance to OECD guideline 471 with minor deviations. Therefore, the results can be considered to be reliable.
in vitro cytogenicity / chromosome aberration study in mammalian cells
In a key chromosomal aberration test (Ishidate, 1984) Chinese hamster lung fibroblast CHO cells were exposed to DL-Alanine in physiological saline in at least three different concentrations up to 1 mg/plate for 24 and 48 hours without metabolic activation. The results were considered to be negative since the incidence of polyploid cells after 48 hr was 1.0% and the incidence of cells with structural aberrations at 24 or 48 hr after treatment was 1.0% as well.
The study was performed in accordance to OECD guideline 473 with minor deviations. Therefore, the results can be considered to be reliable.
Other studies with structurally related amino acids were also negative. Moreover EFSA (2008) concluded that genotoxicity of amino acids, including DL-alanine is not likely.
Justification for classification or non-classification
The substance DL-Alanine was tested negative in a reliable reverse gene mutation test in bacteria (Ames test) with the strains TA 1535, TA 1537, TA 92, TA 98, TA94 and TA 100 of S. typhimurium with metabolic activation. A reliable chromosomal aberration test in Chinese hamster lung fibroblast CHO cells with the test substance was also negative. Therefore no classification as mutagenic according to Regulation (EC) No 1272/2008 (CLP Regulation) is warranted.
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