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EC number: 208-167-3 | CAS number: 513-77-9
- 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
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- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
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- Endpoint summary
- Stability
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- 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 in bacteria (OECD 471): negative
in-vitro gene mutation in mammalian cells (OECD 476): negative
in-vitro chromosome aberration (OECD 473): negative
Link to relevant study records
- 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:
- other: Guideline conform study with GLP
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Species / strain / cell type:
- S. typhimurium TA 97
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- 100, 333, 1000, 3333, 10000 µg/plate, duplicate cultures
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: no data - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-Aminoanthracen (all strains in the presence of S9)
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- TA1535 and TA100
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- TA97 and TA1537
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 4-nitro-o-phenylenediamine (TA98)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: pre-incubation; barium chloride dihydrate was incubated with the Salmonella typhimurium tester strains
either in buffer or S9 mix for 20 minutes at 37°C. Top agar supplemented with I-histidine and d-biotin was added, and the contents of the tubes were mixed and poured onto the surfaces of minimal glucose agar plates.
DURATION
- Exposure duration: 20 minutes
- Expression time (cells in growth medium): 2 days at 37°C
NUMBER OF REPLICATIONS: Each trial consisted of triplicate plates of concurrent positive and negative controls, and of at least five doses of barium chloride dihydrate.
No further details are given. - Evaluation criteria:
- In this test, a positive response was defined as a reproducible, dose-related increase in histidin-independent (revertant) colonies in any one strain/activation combination. An equivocal response was defined as an increase in revertants that was not dose related, not reproducible, nor was of sufficient magnitude to support a determination of mutagenicity. A negative response was obtained when no increase in revertant colonies was observed following chemical treatment. There was no minimum percentage or fold increase required for a chemical to be judged positive or weakly positive.
- Statistics:
- Statistical analysis is not mandatory for the bacterial reverse mutation assay.
- Species / strain:
- S. typhimurium TA 97
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: slight toxicity at concentration from 333 to 10000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: slight toxicity at concentration from 333 to 10000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: slight toxicity at concentration from 333 to 10000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: slight toxicity at concentration from 333 to 10000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: slight toxicity at 10000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- no data
- Conclusions:
- No increased induction of revertant colonies at all concentrations in all strains with and without metabolic activation.
- Executive summary:
Read across from BaCl2*2H2O to BaCO3:
Based on the negative results in thein-vitrotest for the soluble barium compound (BaCl2*2H2O) and assuming that poorly soluble compounds are less bioavailable, read-across from BaCl2*H2O is performed.
- 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:
- other: Guideline conform study with GLP
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- no data available
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-mix
- Test concentrations with justification for top dose:
- Trial 1, without S9: 50, 160, 500 and 1600 µg/mL
Trial 1, with S9: 50, 160, 500, 1600 and 5000 µg/mL
Trial 2, without S9: 100, 250, 500, 1000, 1500 and 2000 µg/mL
Trial 2, with S9: 500, 1600, 3000, 4000 and 5000 µg/mL - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: no data - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium, duplicate cultures
DURATION (test without S9)
- Exposure duration: In the test without S9, cells were incubated with barium chloride dihydrate for 10 hours.
Spindel inhibitor: (cytogenetic assays): Colcemid was added and incubation continued for 2 hours.
Stain: (for cytogenetic assays): The cells were then harvested by mitotic shake-off, fixer, and stained with Giemsa.
DURATION (test with S9)
- Exposure duration: In the test with S9, cells were treated with barium chloride dihydrate and S9 for 2 hours, after which the treatment medium was removed and the cells were incubated for 10 to 11 hours in fresh medium.
Spindel inhibitor: (cytogenetic assays): Colcemid was added for the final 2 hours.
Stain: (for cytogenetic assays): The cells were then harvested by mitotic shake-off, fixer, and stained with Giemsa.
NUMBER OF CELLS EVALUATED: One hundred first-division metaphase cells were scored at each dose level. Classes of aberrations included simple (breaks and terminal deletions), complex (rearrangements and translocations), and other (pulverised cells, despiralised chromosomes, and cells containing 10 or more aberrations).
No further details are given. - Evaluation criteria:
- no data
- Statistics:
- CA data are presented as percentage of cells with aberrations. To arrive at a statistical call for a trial, analyses were conducted on both the dose response curve and individual dose points. For a single trial, a statistically significant (P<0.05) difference for one dose point and a significant trend (P<0.015) are considered weak evidence for a positive response; significant differences for two or more doses indicate the trial is positive. A positive trend test in the absence of a statistically significant increase at any one dose results in an equivocal call (Galloway et al., 1987).
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- no data
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- The in vitro chromosome aberration test in CHO cells was negative with and without metabolic activation.
- Executive summary:
Read across from BaCl2*2H2O to BaCO3:
Based on the negative results in the in-vitro test for the soluble barium compound (BaCl2*2H2O) and assuming that poorly soluble compounds are less bioavailable, read-across from BaCl2*H2O is performed.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 15 December 2009 till 11 February 2010
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: GLP guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- signed by The Department of Health of the Government of the United Kingdom (2010-06-23)
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- tk +/- locus
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- The master stock of L5178Y tk+/- mouse lymphoma cells originated from Dr Donald Clive, Burroughs Wellcome Co.
Cells supplied to Covance Laboratories Ltd. were stored as frozen stocks in liquid nitrogen. The cells were diluted in RPMI 10 and incubated in a humidified atmosphere of 5% (v/v) CO2 in air. When the cells were growing well, subcultures were established in an appropriate number of flasks.
- Type and identity of media: RPMI 1640 media supplemented with heat inactivated horse serum (0%, 10% or 20% (v/v)), 100 units/mL penicillin and 100 µg/mL streptomycin, 2.5 µg/mL Amphotericin B, 0.2 mg/mL pyruvic acid and 0.5 mg/mL pluronic (except fpr RPMI 20%).
- Properly maintained: yes/no
- Periodically checked for Mycoplasma contamination: yes; Each batch of frozen cells was checked that it was mycoplasma free.
- Periodically "cleansed" against high spontaneous background: yes; Each batch of frozen cells was checked for spontaneous mutant frequency. - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- Concentrations selected for Experiments I and II were based on the results of this cytotoxicity Range-Finder Experiment.
Range-Finder:
- with and without S9 mix, 3 hours treatment: 65.06, 130.1, 260.3, 520.5, 1041, 2082 µg/mL
- without S9 mix, 24 hours treatment: 8.133, 16.27, 32.53, 65.06, 130.1, 260.3, 520.5, 1041, 2082 µg/mL
Experiment I:
- with and without S9 mix, 3 hours treatment: 250, 500, 750, 1000, 1200, 1400, 1600, 1800, 1950, 2082 µg/mL
In Experiment I, cultures treated at 1000 and 1200 µg/mL in the absence of S9 mix gave 26% and 4% relative total growth, respectively, therefore Experiment II consisted of 3 hour treatments in the absence and presence of S9 mix and a 24 hour treatment in the absence of S9 mix in order to better define the toxicity profile.
Experiment II:
- without S9 mix, 3 hours treatment: 100, 200, 400, 600, 800, 900, 1000, 1100, 1200, 1400 µg/mL
- with S9 mix, 3 hours treatment: 200, 400, 600, 800, 900, 1000, 1100, 1200, 1300, 1400 µg/mL
- without S9 mix, 24 hours treatment: 100, 200, 400, 600, 800, 900, 1000, 1100, 1200, 1400 µg/mL - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: purified water
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- treatments with the vehicle purified water diluted 10 fold in the treatment medium
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- without metabolic activation Migrated to IUCLID6: Experiment I: 15.0 and 20.0 µg/mL and Experiment II: 5.0 and 7.5 µg/mL; dissolved in DMSO
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- Remarks:
- with metabolic activation Migrated to IUCLID6: 2.0 and 3.0 µg/mL; dissolved in DMSO
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
For Experiments I and II in the absence and presence of S9 mix least 10^7 cells in a volume of 17 mL tissue culture medium were used.
For Experiment II in the absence of S9 mix at least 4 x 10^6 cells in a volume of 18 mL RPMI 10 were used.
DURATION
- Exposure duration: 3 hours (with and without S9 mix; Experiment I and II) and 24 hours (without S9 mix; Experiment II), at 37±1°C
After incubation, cultures were centrifuged, washed and resuspended fresh RPMI 10 medium. Cells were transferred to tissue culture flasks for growth throughout the expression period.
- Expression time (cells in growth medium): Cultures were maintained in flasks for a period of 2 days during which the tk-/- mutation would be expressed. Some cultures were selected for determination of viability and TFT resistance (see table below).
- Fixation time (start of exposure up to fixation or harvest of cells): 12 to 14 days; At the end of the expression period, the cells were placed into each well of four 96 well microtitre plates (384 wells at 2 x 10³ cells/well). Plates were incubated at 37±1°C with 5% v/v CO2 in air until scoreable and wells containing clones were identified.
SELECTION AGENT (mutation assays): TFT (5-trifluorothymidine)
NUMBER OF REPLICATIONS: duplicate cultures (single cultures only used for positive control treatments)
NUMBER OF CELLS EVALUATED: Number of cells per well is 2000 cells per well on average on all mutant plates.
EVALUATION: Mutant frequency (MF) was calculated as follows:
MF = -ln P(o) for mutant plates/Number of cells per well x (viability/100)
DETERMINATION OF CYTOTOXICITY
- Method: viability, cloning efficiency, relative total growth:
In the absence of S9 mix, 3 and 24 hours treatment incubation periods were used, in the presence of S9 mix a 3 hour treatment incubation was used.
Following treatment, cells were centrifuged, washed with tissue culture medium and resuspended in RPMI 10. Cell densities were determined. All cultures were incubated at 37±1°C for 1 day, recounted and where possible diluted to 2 x 10^5 cells/mL. Cultures were incubated for a further day, counted and adjusted to 8 cells/mL and, for each concentration, 0.2 mL was plated into each well of a 96 well microtitre plate (incubation at 37±1°C with 5% CO2 in air for 7 to 11 days) for determination of viability (plating efficiency). Wells containing viable clones were identified by eye. Viability is the measure of the cells' ability to clone i.e. Cloning efficiency (CE).
Relative Total Growth (RTG) is the measure of cytotoxicity relative to the control, that takes into account all cell growth, cell loss during the treatment period and the 2 day expression period (RSG) and the cells' ability to clone 2 days after treatment (viability). - Evaluation criteria:
- For valid data, the test article was considered to be mutagenic in this assay if:
1. The mutation frequency (MF) of any test concentration exceeded the sum of the mean control mutant frequency plus global evaluation factor (GEF)
2. The linear trend test was positive.
The test article was considered as positive in this assay if both of the above criteria were met.
The test article was considered as negative in this assay if neither of the above criteria were met.
Results which only partially satisfied the assessment criteria described above were considered on a case-by-case basis. - Statistics:
- The significance of increases in mutant frequencies (total wells with clones), by comparison with concurrent controls and the global evaluation factor (GEF), was assessed according to the recommendations of the Mouse Lymphoma Workgroup, Aberdeen, 2003. The control mutant frequency was compared with each test article treatment and the data were checked for a linear trend in mutant frequency with treatment concentration using weighted regression. The test for linear trend is one-tailed, therefore negative trend was not considered significant.
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- The mutant frequency of the concentrations plated were all less than the sum of the mean control mutant frequency plus the Global Evaluation Factor (GEF, 126 mutants per 10^6 viable cells), indicating a negative result. (For details see sttached document)
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No marked changes in pH were observed in the 3 and 24 hours Range Finder Experiments at the highest concentration tested.
- Effects of osmolality: No marked changes in osmolality were observed in the 3 and 24 hours Range Finder Experiments at the highest concentration tested.
- Water solubility: Preliminary solubility data indicated that Barium chloride dihydrate was soluble in water for irrigation (purified water) at concentrations up to at least 23.85 mg/mL (anhydrous). The solubility limit in culture medium was below 2385 µg/mL (anhydrous), as indicated by the appearance of slight precipitate (haziness) at this concentration approximately 26 hours after test article addition. A maximum concentration of 2082 µg/mL (anhydrous) was selected for the cytotoxicity Range Finder Experiment in order that treatments were performed up to 10 mM.
RANGE-FINDING/SCREENING STUDIES: In the cytotoxicity Range-Finder Experiment, 3 hour treatment, 6 concentrations were tested, in the absence and presence of S 9, ranging from 65.06 to 2082 µg/mL (equivalent to 10 mM anhydrous Barium chloride dihydrate at the highest concentration tested). The highest concentration, 2082 µg/mL, gave 18% and 31% relative total growth (RTG) in the absence and presence of S9 mix, respectively.
In the cytotoxicity Range-Finder Experiment, 24 hour treatment, 9 concentrations were tested in the absence of S9 mix, ranging from 8.133 to 2082 µg/mL. The highest concentration to provide >10% RTG was 1041 µg/mL, which gave 24% RTG.
COMPARISON WITH HISTORICAL CONTROL DATA: no data
ADDITIONAL INFORMATION ON CYTOTOXICITY: no further data - Conclusions:
- Interpretation of results (migrated information):
negative
It is concluded that Barium chloride dihydrate did not induce mutation at the tk locus of L5178Y mouse lymphoma cells when tested under the conditions employed in this study. These conditions included treatments up to toxic and/or precipitating concentrations in two independent experiments in the absence and presence of a rat liver metabolic activation system (S9 mix). - Executive summary:
Read across from BaCl2*2H2O to BaCO3:
Based on the negative results in the in-vitro tests for the soluble barium compound (BaCl2*2H2O) and assuming that poorly soluble compounds are less bioavailable, read-across from BaCl2*H2O is performed.
Conclusion:
Barium chloride dihydrate was assayed for its ability to induce mutation at the tk locus (5‑trifluorothymidine [TFT] resistance) in mouse lymphoma cells.
Experiment I was performed using a 3 -hour treatment incubation and Experiment II was performed using 3- and 24 -hour treatment incubations.
In the cytotoxicity Range-Finder Experiment, concentrations were tested ranging from 65.06 to 2082 µg/mL (3 -hour treatment; -/+S9 mix) and from 8.133 to 2082 µg/mL (24 -hour treatment; -S9 mix).
Concentrations selected for Experiments I and II were based on the results of this cytotoxicity Range-Finder Experiment.
Two Experiments were performed with selected concentrations:
- In Experiment I concentrations, ranging from 250 to 2082 µg/mL,were tested in the absence and presence of S9 mix.
- In Experiment II (3-hour treatment) concentrations, ranging from 100 to 1400 µg/mL in the absence of S9 mix and from 200 to 1400 µg/mL in the presence of S9 mix, were tested. Additionally cells were tested in a 24 -hour treatment with concentrations ranging from 100 to 1400 µg/mL in the absence of S9 mix.
Negative (vehicle) and positive control treatments were included in each Mutation Experiment. Mutant frequencies in negative control cultures fell within acceptable ranges, and clear increases in mutation were induced by the positive control chemicals Methyl methane sulphonate (without S‑9) and Benzo[a]pyrene (with S‑9). Therefore the study was accepted as valid.
In Experiments I and II, the mutant frequency of the concentrations plated were all less than the sum of the mean control mutant frequency plus the Global Evaluation Factor, indicating a negative result. Statistically significant linear trends were observed in Experiment II in the presence of S9 mix (3-hour treatment) and in the absence of S9 mix (24-hour treatment). However, in the absence of any marked increases in mutant frequency under either treatment condition, these observations were not considered biologically relevant.
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
Read-across BaCl2*2H2O to BaCO3:
Based on the negative results in the in-vitro tests for soluble barium compounds (BaCl2*2H2O) and assuming that poorly soluble compounds are less bioavailable, read-across from BaCl2*H2O is performed.
Justification for classification or non-classification
None of the in vitro genotoxicity studies rated as reliable showed any effect in bacterial reverse mutation assays, in mammalian cell gene mutation tests (TK assay) or in mammalian cell chromosome aberration tests, thus the classification criteria according to regulation (EC) 1272/2008 as germ cell mutagen are not met.
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