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EC number: 237-149-8 | CAS number: 13669-76-6
- 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
![](https://echa.europa.eu./o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- From 2010-01-25 to 2010-03-01
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Well documented GLP study performed according to the UK Environmental Mutagen Society which is based on the OECD Guideline 476 and EU Method B.17. There was only a minor deviation reported: no analysis was performed on homogeneity, concentration or stability of the test material formulation, however this is considered not to affect the purpose or integrity of the study.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 010
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- other: United Kingdom Environmental Mutagen Society
- Deviations:
- yes
- Remarks:
- no analysis on homogeneity, concentration or stability of test material formulation, however this is considered not to affect the purpose or integrity of the study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- yes
- Remarks:
- no analysis on homogeneity, concentration or stability of test material formulation, however this is considered not to affect the purpose or integrity of the study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- yes
- Remarks:
- no analysis on homogeneity, concentration or stability of test material formulation, however this is considered not to affect the purpose or integrity of the study
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
Test material
- Reference substance name:
- Orthophosphoric acid
- EC Number:
- 231-633-2
- EC Name:
- Orthophosphoric acid
- Cas Number:
- 7664-38-2
- IUPAC Name:
- phosphoric acid
- Details on test material:
- - Name of test material: phosphoric acid 85%
- Substance type: inorganic
- Physical state: clear colourless liquid
- Analytical purity: 85.6%
- Purity test date: 2010-01-06
- Lot/batch No.: 209
- Storage condition of test material: at room temperature in the dark
Constituent 1
Method
Species / strain
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Cells were routinely cultured in RPMI 1640 medium with Glutamax-1 and HEPES buffer (20 mM) supplemented with penicillin (100 units/mL), streptomycin (100 µg/mL), sodium pyruvate (1 mM), amphotericin B (2.5 µg/mL) and 10% donor horse serum (giving R10 media) at 37°C with 5% CO2 in air.
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: no
- Periodically checked for karyotype stability: no
- Periodically "cleansed" against high spontaneous background: yes: before the stocks of cells were frozen they were cleansed of homozygous (TK -/-) mutants by culturing in THMG medium for 24 hours. This medium contained thymidine (9 µg/mL), hypoxanthine (15 µg/mL), methotrexate (0.3 µg/mL) and glycine (22.5 µg/mL). For the following 24 hours the cells were cultured in THG medium (i.e. THMG without methotrexate) before being returned to R10 medium.
- Metabolic activation:
- with and without
- Metabolic activation system:
- phenobarbitol/beta-naphthoflavone S9 from livers of male Sprague-Dawley rats
- Test concentrations with justification for top dose:
- 0, 61.25, 122.5, 245, 490, 735 and 980 µg/mL
- Vehicle / solvent:
- - Solvent(s) used: R0 medium
Controlsopen allclose all
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- in the absence of metabolic activation
Migrated to IUCLID6: Sigma batch 142314732109252 at 400 µg/mL and 150 µ/mL for the 4 - and 24-hour exposures respectively
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- in the presence of metabolic activation
Migrated to IUCLID6: Acros batch A0164185 at 2 µg/mL
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: not applicable
- Exposure duration: 4-hour exposure with and without metabolic activation; 24-hour exposure without metabolic activation
- Expression time (cells in growth medium): 2 days (dilution of 2 x 1E05 cells/mL)
- Selection time (if incubation with a selection agent): 10 to 14 days of incubation
- Fixation time (start of exposure up to fixation or harvest of cells): 3 hours (0.025 mL of MTT solution, 2.5 mg/mL in PBS)
SELECTION AGENT (mutation assays): selective medium containing 4 µg/mL 5-trifluorothymidine (TFT in 96-well microtitre plates
SPINDLE INHIBITOR (cytogenetic assays): not applicable
STAIN (for cytogenetic assays): not applicable
NUMBER OF REPLICATIONS: duplicates
NUMBER OF CELLS EVALUATED: At the end of the treatment period, for each experiment, the cells were washed twice using the R10 medium then resuspended in R20 medium at a cell density of 2 x 1E05 cells/mL. The cultures were incubated and subcultured every 24 hours for the expression period of two days, by counting and dilution to 2 x 1E05 cells/mL. On day 2 of the experiment, the cells were counted, diluted to 1E04 cells/mL and plated for mutant frequency (2000 cells/well) in selective medium containing 4 µg/mL 5-trifluorothymidine (TFT) in 96-well microtitre plates. Cells were also diluted to 10 cells/mL and plated (2 cells/well) for viability (% V) in non-selective medium. The daily cell counts were used to obtain a Relative Suspension Growth (%RSG) value that gives an indication of post treatment toxicity during the expression period as a comparison to the vehicle control, and when combined with the Viability (% V) data a Relative Total Growth (RTG) value.
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency, viability, relative suspension growth and relative total growth
OTHER EXAMINATIONS:
- Determination of polyploidy: no
- Determination of endoreplication: no
- Other: no - Evaluation criteria:
- The normal range for mutant frequency per survivor is 50-200 x 1E-06 for the TK +/- locus in L5178Y cells at Harlan Laboratories Ltd. Vehicle controls results should ideally be within this range, although minor errors in cell counting and dilution or exposure to the metabolic activation system may cause this to be slightly elevated. Experiments where the vehicle control values are markedly greater than 250 x 1E-06 mutant frequency per survivor are not normally acceptable and will be repeated. Positive control chemicals should induce at least three to five fold increases in mutant frequency greater than the corresponding vehicle control. For a test material to demonstrate a mutagenic response it must produce a statistically significant increase in the induced mutant frequency (IMF) over the concurrent vehicle mutant frequency value. The IMF must exceed 126 x 1E-06 for the microwell method based on the global background MF (Moore et al 2003). Therefore any test material dose level that has a mutation frequency value that is greater than the corresponding vehicle control by the GEF of 126 x 1E-06 will be considered positive. However, if a test material produces a modest increase in mutant frequency, which only marginally exceeds the GEF value and is not reproducible or part of a dose-related response, then it may be considered to have no toxicological significance. Conversely, when a test material induces modest reproducible increases in the mutation frequencies that do not exceed the GEF value then scientific judgement will be applied. If the reproducible responses are significantly dose-related and include increases in the absolute numbers of mutant colonies then they may be considered to be toxicological significant.
- Statistics:
- The experimental data were analysed using a dedicated computer program which follows the statistical guidelines recommended by the UKEMS. Dose levels that have survival values less than 10% are excluded from any statistical analysis, as any response they give would be considered to have no biological or toxicological relevance.
Results and discussion
Test resultsopen allclose all
- Species / strain:
- other: L5178Y TK+/- 3.7.2c mouse lymphoma cell line
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- 4-hour exposure with and without metabolic activation
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Very modest dose related toxicity, no evidence of any reductions in viability (%V), therefore no residual toxicity occurred
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- other: L5178Y TK+/- 3.7.2c mouse lymphoma cell line
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Remarks:
- 24-hour exposure without metabolic activation
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Modest dose related toxicity following exposure to the test material. There was no evidence of any reductions in viability (%V), therefore indicating that no residual toxicity had occurred.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no data on effects, however it was mentioned that there was no marked change in pH when the test material was dosed into media
- Effects of osmolality: no data on effects, however it was mentioned that the osmolality did not increase by more than 50 mOsm
- Precipitation: no precipitate of the test material was observed at any of the dose levels tested in the preliminary toxicity test (0, 3.83, 7.66, 15.31, 30.63, 61.25, 122.5, 245, 490, 980)
RANGE-FINDING/SCREENING STUDIES: In the three exposure groups (4-h exposure -S9, 4-hour exposure +S9 and 24-hour exposure -S9) there were no marked dose related reductions in the Relative Suspension Growth (%RSG) of treated cells with the test material when compared to the concurrent vehicle controls. However, a modest reduction was observed at the 10 mM limit dose in all three of the exposure groups with the greatest reduction observed in the 24-hour exposure group in the absence of metabolic activation. No precipitate of the test material was observed at any of the dose levels. In the subsequent mutagenicity test, the maximum dose level for all three of the exposure groups was the 10 mM limit dose.
COMPARISON WITH HISTORICAL CONTROL DATA: The mutation frequency is situated within the range of historical control values for the positive controls and solvent control.
ADDITIONAL INFORMATION ON CYTOTOXICITY: no data - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
Results for 4 -hour exposure (-S9):
%RSG for treatment: ranged from 110 to 75 (from dose 61.25 to 980 µg/mL)
%V for treatment: ranged from 84.40 to 90.38 (from dose 61.25 to 980 µg/mL)
RTG for treatment: ranged from 1.10 to 0.80 (from dose 61.25 to 980 µg/mL)
MF for treatment: ranged from 97.01 to 82.17 (from dose 61.25 to 980 µg/mL)
The values for the solvent control were 100 (%RSG), 84.40 (%V), 1.00 (RTG) and 84.42 (MF)
The values for the positive control EMS were 76 (%RSG), 88.81 (%V), 0.80 (RTG) and 688.67 (MF)
Results for 4 -hour exposure (+S9):
%RSG for treatment: ranged from 94 to 81 (from dose 61.25 to 980 µg/mL)
%V for treatment: ranged from 95.38 to 96.26 (from dose 61.25 to 980 µg/mL)
RTG for treatment: ranged from 1.07 to 0.92 (from dose 61.25 to 980 µg/mL)
MF for treatment: ranged from 107.17 to 86.65 (from dose 61.25 to 980 µg/mL)
The values for the solvent control were 100 (%RSG), 84.40 (%V), 1.00 (RTG) and 115.46 (MF)
The values for the positive control CP were 53 (%RSG), 39.58 (%V), 0.25 (RTG) and 1311.27 (MF)
Results for 24 -hour exposure (-S9):
%RSG for treatment: ranged from 104 to 53 (from dose 61.25 to 980 µg/mL)
%V for treatment: ranged from 113.09 to 97.17 (from dose 61.25 to 980 µg/mL)
RTG for treatment: ranged from 1.28 to 0.57 (from dose 61.25 to 980 µg/mL)
MF for treatment: ranged from 55.11 to 70.25 (from dose 61.25 to 980 µg/mL)
The values for the solvent control were 100 (%RSG), 91.99 (%V), 1.00 (RTG) and 79.09 (MF)
The values for the positive control EMS were 55 (%RSG), 85.83 (%V), 0.52 (RTG) and 692.36 (MF)
Applicant's summary and conclusion
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.
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