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EC number: 231-634-8 | CAS number: 7664-39-3
- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
HF: OECD 471 - Negative
NaF (read-across analogue): OECD 473 - Inconclusive
NaF (read-across analogue): OECD 476 - Negative & Positive studies
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 1996
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Published study.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- yes
- Remarks:
- : no metabolic activation
- GLP compliance:
- not specified
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- HGPRT
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- Cells were cultured in MEM with the addition of 6% fetal calf serum, penicillin (200U/ml), streptomycin (100µg/ml), a mixture of non-essential amino acids, and 0.1g/l sodium pyruvate. Cells were maintained in a humidified 5% carbon dioxide atmosphere at 37oC in glass Petri dishes. Cells were checked under an electron microscope for the presence of mycoplasma: no contamination was noted.
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- without
- Test concentrations with justification for top dose:
- Various concentrations between 0 and 500µg/ml at neutral pH or pH4.
- Vehicle / solvent:
- Phosphate buffered saline (PBS).
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- PBS
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: N-Methyl-N-nitro-N-nitrosoguanidine (MNNG)
- Remarks:
- MNNG dissolved in DMSO and diluted in PBS.
- Details on test system and experimental conditions:
- Cytotoxicity assay: Cells were exposed to the substance dissolved in PBS adjusted to either pH4 or pH7 for 30 minutes (0, 200, 300, 400 and 500µg/ml). Controls were kept in PBS at pH4 or 7 for 30 minutes. The cells were aspirated, rinsed with MEM and number of cells counted at 24 hourly intervals. Normal pH media were used after exposures.
6-thioguanine-resistant (6-TG) mutations: 1x10exp6 cells were plated in 10cm Petri dishes and cultivated for 24 hours. The medium was removed and replaced with different concentrations (0, 100, 125, 150 and 200µg/ml) of NaF dissolved in PBS adjusted to pH 4 for 60 minutes, or with MNNG solution (1µg/ml) in PBS buffer at pH7 for 30 minutes. - Evaluation criteria:
- Cytoxocity: cells counted at 24 hour internals.
6-TG mutations: The number of -TG resistant mutations was determined using the 'respreading' method of Chasin (1973). The expression time was 8 days. - Statistics:
- One way ANOVA was used to evaluate gene mutation, and least-significance difference test.
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Cytotoxic effects higher at lower pH
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- At neutral pH, NaF concentrations or 300µg/ml or more inhibited cell growth for 24 hour following treatment; cells treated with 200, 300, 400 and 500 µg/ml proliferated similarly to controls after 24 hours. In cells treated with NaF at pH4, a larger toxic effect was observed. Inhibition of cell growth was observed during the first 24 hours after treatment even at the lowest concentration (200µg/ml). Growth was inhibited in cells treated with 300µg/ml beyond 24 hours since treatment, and cells treated with 400 and 500µg/ml did not survive. Incubation of cells at reduced pH reduced the level of spontaneous 6-TG mutations. No other significant differences were observed.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
other: Cytotoxic at low pH, not mutagenic.
Sodium fluoride was highly toxic, but not mutagenic. - Executive summary:
The mutagenic activity of sodium fluoride was tested in neutral and acidic conditions in the V79/HGPRT system. Sodium fluoride was found to be cytotoxic at neutral pH, and highly cytotoxic in acidic conditions. Sodium fluoride treatment did not result in any mutagenic activity, and incubatuin of cells at reduced pH reduced the level of spontaneous mutations. The authors suggest that an acid environment which supports formation of hydrogen fluoride increases toxic but not mutagenic potencies of sodium fluoride.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 1990
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: NTP study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- GLP compliance:
- not specified
- Type of assay:
- mammalian cell gene mutation assay
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 prepared from the livers of either Aroclor 1254-induced or noninduced Fischer 344 male rats.
- Test concentrations with justification for top dose:
- Two laboratories performed the test. The highest dose of the study compound did not exceed 5 mg/mL.
- Vehicle / solvent:
- No vehicle used
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Details on test system and experimental conditions:
- Mouse lymphoma L5178Y cells were maintained at 37 degC as suspension cultures in Fischer's medium supplemented with 2 mM l-glutamine, 110 µg/mL sodium pyruvate, 0.05% pluronic F68, antibiotics and heat activated horse serum. To reduce the number of spontaneously occurring trifluorothymidine (TFT) resistant cells, subcultures were exposed once to medium containing THMG (thymidine, hypoxanthine, methotrexate, glycine) for one day, to THG for one day and to normal medium for 3 to 5 days.
Incubation with the study chemical continued for 4 hours, at which time the medium plus chemical was removed. The cells were resuspended in fresh medium and incubated for an additional 2 days to allow expression of the mutant phenotype. Cell density was monitored so that log phase growth was maintained. After the 48 hour expression period, 3 x 10(+6) cells were plated in medium and soft agar supplemented with trifluorothymidine for selection of TFT-resistant cells and 600 cells were plated in nonselective medium and soft agar to determine cloning efficiency. Plates were incubated at 37 degC in 5% carbon dioxide for 10 to 12 days. - Evaluation criteria:
- All data was evaluated statistically for both trend and peak response. Both responses had to be significant (P<0.05) for a chemical to be considered positive and the absence of both trend and a peak response resulted in a negative call. Validation criteria are detailed in Myhr at al (1985). This assay is initially performed without S9; if a clearly positive response is not obtained, the experiment is repeated with induced S9.
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Two laboratories were used to test the substance. In the first laboratory, sodium fluoride was positive both with and without Aroclor 1254-induced male Fischer 344 rat liver S9; the effective doses, with and without S9, ranged from 300 to 600 µg/mL. In the second laboratory, sodium fluoride was tested without S9 only and test results were positive in the first trial at 62.5, 125 and 1,000 µg/mL and in the second trial at 800 and 900 µg/mL.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
positive
Sodium fluoride was found to be positive with and without metabolic activation at various concentrations. - Executive summary:
Two laboratories were used to test the substance. In the first laboratory, sodium fluoride was positive both with and without Aroclor 1254-induced male Fischer 344 rat liver S9; the effective doses, with and without S9, ranged from 300 to 600 µg/mL. In the second laboratory, sodium fluoride was tested without S9 only and test results were positive in the first trial at 62.5, 125 and 1,000 µg/mL and in the second trial at 800 and 900 µg/mL.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1990
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: NTP study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Principles of method if other than guideline:
- Assessment of Sister Chromatid Exchange (SCE) and chromosomal aberration.
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- Not relevant
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254-induced male Sprague-Dawley rat liver S9 and cofactor mix.
- Test concentrations with justification for top dose:
- In each trial, 5 different concentrations of sodium fluoride were tested in the range 1.6 to 1600 µg/mL.
- Vehicle / solvent:
- No vehicle used.
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Details on test system and experimental conditions:
- Sodium chloride was tested in cultured Chinese Hamster ovary (CHO) cells for induction of sister chromatid exchanges (SCE) and chromosomal aberrations (Abs) both in the presence and absence of Aroclor of Aroclor 1254 induced male Sprague-Dawley rat liver S9 and cofactor mix. Cultures were handled under gold lights to prevent photolysis of bromodeoxyuridine substituted DNA. Below gives the test system ad conditions used for each trial:
1. In the SCE test without S9, CHO cells were incubated for 26 hours with the study chemical in McCoy's 5A medium supplemented with 10% fetal bovine serum, l-glutamine (2 mM) and antibiotics Bromodeoxyuridine (BrdU) was added 2 hours after culture initiation. After 26 hours, the medium containing the chemical was removed and replaced with fresh medium plus BrdU and Colcemid and incubation was continued for 2 more hours. Cells were then harvestedby mitotic shake off, fixed and stained with Hoechst 33258 and Giemsa.
2. In the SCE test with S9, cells were incubated with the chemical, serum-free medium and S9 for 2 hours. The medium was then removed and replaced with medium containing BrdU and no test chemical and incubation continued for an additional 26 hours, with Colcemid present for the final 2 hours. Harvesting and staining was the same for cells treated without S9.
3. In the Abs test without S9, cells were incubated in McCoy's 5A medium with the study chemical for 8 hours; Colcemid was added and incubation continued for 2 hours. The cells were harvested by mitotic shake off, fixed and stained with Giemsa.
4. In the Abs test with S9, cells were treated with the study chemical and S9 for 2 hours, after which the treatment medium was removed and the cells incubated for 10 hours in fresh medium with Colcemid present for the final 2 hours. Cells were harvested in the same manner as for treatment without S9. - Evaluation criteria:
- Statistical analysis were conducted on both the slopes of the dose response curves and the individual dose points. An SCE frequency 20% above the concurrent solvent control value was chosen as a statiscally conservative positive response. If one dose point was positive, the chemical was termed 'weak positive'; if two or more dosesw ere positive, the chemical was judged 'positive.' Abs data was presented as percentage of cells with aberrations. As with SCE, both the dose-response curve and individual dose points were statistically analysed. A statistically significant (P<0.003) effect on the slope of the curve or on at least two dose points (P<0.05) was sufficient for a conclusion of positive for a test.
- Statistics:
- See above in evaluation criteria.
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- ambiguous
- Remarks:
- : see discussion below
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
ambiguous
The results are inconclusive. The two laboratories used to test the effects of sodium fluoride on CHO cells showed conflicting results; one reported a negative result and one reported a positive result for both induction of sister chromatid exchanges (SCE) and chromosomal aberrations (Abs). - Executive summary:
The results are inconclusive. The two laboratories used to test the effects of sodium fluoride on CHO cells showed conflicting results; one reported a negative result and one reported a positive result for both induction of sister chromatid exchanges (SCE) and chromosomal aberrations (Abs).
- 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:
- July - August 1987
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- no data concerning analytical investigations of the test substance were available, and the content given was based on a representative sample. This small deviation is not expected to have affected the results
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- No further information.
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- other: histidine-deficient
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix made from the livers of rats injected with Aroclor 1254 5 days prior to sacrifice
- Test concentrations with justification for top dose:
- Nominal test concentrations in the first experiment were 12500, 2500, 500, 100, 20 and 0 µg per plate. Nominal test concentrations in the second experiment were 1800, 900, 450, 225, 112.5, 56.3 and 0 µg per plate.
- Vehicle / solvent:
- The solvent for hydrogen fluoride was deionised water, and DMSO for the positive controls.
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- TA 1535
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: nitrofurantoin
- Remarks:
- TA 100
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 4-nitro-1,2-phenylene diamine
- Remarks:
- TA 1573 and TA 98
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- Control for the activation activity of S9
- Details on test system and experimental conditions:
- Immediately on receipt, the samples were inoculated on nutrient agar plates, to which ampicillin had been added in the case of TA 100 and TA 98. These plates were incubated at 37°C for appproximately 24 hours. With a sterile inoculation loop samples were taken from individual colonies and transferred to nutrient broth. Ampicillin was added to the broth in the case of TA 100 and TA 98. The samples were incubated overnight at 37°C. New samples of these cultures were inoculated on to nutrient agar plates, which had again been provided with ampicillin for TA 100 and TA 98. After an incubation period of approximately 24 hours at 37°C, new samples of individual colonies of these plates were transferred to normal nutrient broth, which was incubated overnight at 37°C, the volume was approx. 30 ml. After incubation a small sample was taken and used to check the genotype. At the same time, the remaining cultures were provided with DMSO to protect against freezing and immediately frozen at -80°C in 1 ml portions. Per test and strain one of the 1 ml portions was thawed, and quantities of 0.2 ml thawed culture were added to 10 ml nutrient broth, then incubated overnight at 37°C.
A test for crystal violet sensitivity and a test for UV sensitivity were made. Cultures not producing satisfactory data at this point were discarded.
Four plates per strain, substance and dose both with and without S9 mix were used. The same number of plates made up the negative control (i.e. solvent without test substance). The bacterial suspensions used were obtained from 17 hour cultures in nutrient broth which had been incubated at 37°C and 90 rpm. The count was made after the plates had been incubated for 48 hours at 37°C. - Evaluation criteria:
- Background growth on the plates was visually assessed. The mutant count per plate was compared to the negative control count. The titre was also determined. A reproducible and dose-related increase in mutant counts for at least one strain was considered positive, and about double the negative control count should be reached. In the case of no reproducible and dose-related increase in mutant counts in at least one strain, the result was evaluated as negative.
- Statistics:
- Not performed.
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- Negative with and without S9 activation.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative Negative with and without S9 activation.
Hydrogen fluoride was negative for mutagenic effects both with and without S9 activation. - Executive summary:
The genotoxicity of hydrogen fluoride 71/75% was determined in Salmonella typhimurium in a standard Ames test both with and without S9 activation. The Salmonella mutants tested were the histidine-auxotrophic strains TA 1535, TA 100, TA 1537 and TA 98. The substance was tested up to a concentration of 12500 µg per plate. Hydrogen fluoride 71/75% was toxic to the bacteria in a strain specific manner at all concentrations, so that the tested range could only be used to a limited extent up to 1800 µg per plate. Substance precipitation occurred at 450 µg per plate. There was no evidence of mutagenic activity at any concentration tested with and without metabolic activation.
Referenceopen allclose all
At neutral pH, NaF concentrations or 300µg/ml or more inhibited cell growth for 24 hour following treatment; cells treated with 200, 300, 400 and 500µg/ml proliferated similarly to controls after 24 hours. In cells treated with NaF at pH4, a larger toxic effect was observed. Inhibition of cell growth was observed during the first 24 hours after treatment even at the lowest concentration (200µg/ml). Growth was inhibited in cells treated with 300µg/ml beyond 24 hours since treatment, and cells treated with 400 and 500µg/ml did not survive. Incubation of cells at reduced pH reduced the level of spontaneous 6-TG mutations. No other significant differences were observed.
The table below shows the induction of trifluorothymidine resistance in mouse L5178Y cells by sodium fluoride;
Compound |
Concentration (µg/mL) |
Cloning Efficiency (%) |
Relative Total Growth (%) |
Tft Resistant Cells |
Mutant fraction |
-S9 Trial 1 Distilled water |
|
75.8 ±3.8 |
99.8 ±4.8 |
67.0 ±5.9 |
29.5 ±1.9 |
Sodium fluoride |
200 300 400 500 600 800 |
85.5 ±18.5 85.3 ±4.7 78.7 ±0.9 75.0 ±3.8 79.5 ±3.5 Lethal |
81.5 ±8.5 72.0 ±5.2 41.0 ±4.0 16.7 ±2.7 10.0 ±1.0 |
80.5 ±13.5 133.3 ±22.3 107.7 ±4.9 125.0 ±16.7 196.0 ±3.0 |
32.0 ±2.0 52.7 ±9.4 45.7 ±1.9 55.3 ±4.8 83.0 ±5.0 |
Methyl methanesulfonate |
5 |
66.7 ±2.4 |
70.7 ±5.2 |
489.0 ±43.7 |
244.3 ±13.2 |
-S9 Trial 2 Distilled water |
|
105.7 ±9.1 |
100.0 ±9.1 |
77.0 ±2.5 |
24.3 ±1.9 |
Sodium fluoride |
50 100 200 300 400 500 600 |
91.0 ±7.0 92.0 ±4.0 88.0 ±4.6 89.3 ±10.2 104.0 ±12.0 94.3 ±7.6 Lethal |
86.3 ±2.0 71.5 ±8.5 57.0 ±2.3 49.3 ±3.7 40.0 ±0.0 17.3 ±3.0 |
59.0 ±9.3 75.5 ±3.5 72.7 ±11.1 58.3 ±8.7 112.0 ±25.0 119.0 ±28.4 |
22.0 ±3.1 27.0 ±0 27.3 ±3.0 22.0 ±2.7 35.5 ±4.5 41.3 ±8.5 |
Methyl methanesulfonate |
5 |
66.7 ±2.4 |
70.7 ±5.2 |
489.0 ±43.7 |
140 ±36.5 |
+S9 Trial 3 Distilled water |
|
107.5 ±1.3 |
100.0 ±6.8 |
83.8 ±6.9 |
25.8 ±2.1 |
Sodium fluoride |
100 200 300 400 500 600 |
85 99.7 ±7.8 94.5 ±5.5 106.7 ±7.0 72.3 ±9.2 77.5 ±13.5 |
75 70.7 ±4.9 52.0 ±9.0 41.3 ±2.3 13.3 ±3.8 7.5 ±0.5 |
66 59.7 ±9.9 110.5 ±5.5 121 ±14.1 177.7 ±45.1 206.5 ±24.5 |
26 20.7 ±4.4 39.5 ±4.5 39.0 ±7.6 81.0 ±19.3 94.0 ±27.0 |
Methyl methanesulfonate |
2.5 |
61.3 ±5.2 |
28.0 ±2.1 |
615.0 ±43.3 |
334.7 ±11.9 |
+S9 Trial 3 Distilled water |
|
82.3 ±3.0 |
100.0 ±9.3 |
81.5 ±8.7 |
33.0 ±2.4 |
Sodium fluoride |
50 100 200 300 400 500 600 |
78.0 ±2.9 79.3 ±6.0 85.0 ±13.1 76.3 ±0.3 77.3 ±3.8 74.3 ±3.4 Lethal |
100.0 ±7.8 86.7 ±21.1 83.3 ±6.4 49.7 ±3.3 29.0 ±1.5 21.0 ±3.1 |
63.7 ±4.2 88.7 ±14.0 98.0 ±18.6 119.0 ±11.8 144.7 ±10.3 167.7 ±27.9 |
27.0 ±1.2 37.7 ±6.8 38.0 ±2.0 51.7 ±5.2 62.3 ±2.3 75.7 ±14.3 |
Methyl methanesulfonate |
2.5 |
47.7 ±2.7 |
21.0 ±2.7 |
731.7 ±22.9 |
573.7 ±34.6 |
The table below shows the results obtained for the induction of trifluorothymidine resistance in mouse L5178Y/TK cells by sodium fluoride;
Compound |
Concentration (µg/mL) |
Cloning Efficiency (%) |
Relative Total Growth (%) |
Tft Resistant Cells |
Mutant fraction |
-S9 Trial 1 Fischer’s medium without serum |
|
58.8 ±3.0 |
100.0 ±10.3 |
101 ±14.0 |
58.0 ±9.7 |
Sodium fluoride |
62.5 125 250 500 1000 |
52.5 ±5.5 52.5 ±8.5 59.0 ±4.0 70.0 ±15.0 40.0 ±4.0 |
88.5 ±12.5 78.0 ±4.0 70.0 ±9.0 36.0 ±1.0 8.0 ±3.0 |
162.0 ±0.0 144.0 ±24.0 130.0 ±9.0 161.5 ±18.5 155.5 ±13.5 |
104.5 ±11.5 97.5 ±31.5 73.5 ±0.5 82.5 ±26.5 134.0 ±25.0 |
Methyl methanesulfonate |
15 |
16.5 ±1.5 |
13.0 ±2.0 |
172.0 ±3.0 |
342.5 ±24.5 |
-S9 Trial 2 Fischer's medium without serum |
|
90.5 ±5.72 |
100.0 ±5.3 |
138.3 ±12.8 |
51.0 ±4.1 |
Sodium fluoride |
500 600 700 800 900 1000 |
82.0 ±3.0 87.0 ±10.0 90.5 ±3.5 78.5 ±0.5 78.0 ±2.0 Lethal |
33.0 ±0.0 28.0 ±3.0 25.0 ±0.0 20.0 ±1.0 13.0 ±1.0 |
145.0 ±20.0 148.5 ±0.5 177.0 ±25.0 215.5 ±22.5 445.0 ±22.0 |
58.5 ±5.5 58.0 ±7.0 66.0 ±12.0 91.5 ±9.5 195.5 ±3.5 |
Methyl methanesulfonate |
1552.0 ±4.0 |
52.0 ±4.0 |
35.0 ±1.0 |
77.0 ±14.0 |
49.0 ±5.0 |
Conflicting results were found in the two laboratories testing sodium fluoride on CHO cells for any cytogenetic effects. SCEs were induced in one laboratory at doses of 66.7 and 75 µg/mL without S9 and at doses of 1200 µg/mL and higher with S9. In two of the five cases, the positive results were seen following delayed harvest to allow cells, whose division time was inhibited by the higher doses of sodium fluoride to progress to the second metaphase division to the point where the cells could be scored. The laboratory reporting negative SCE results did not employ extended harvest times and was able to test up to only 50 µg/mL sodium fluoride without S9 and 500 µg/mL with S9. In the tests for the induction of Abs, positive results were reported in one laboratory at doses of 400 µg/mL sodium fluoride and greater without S9. The second laboratory reported negative results without S9, but the highest dose tested was 200 µg/mL. Neither laboratory showed a reproducible increase in chromosomal aberration in the presence of S9.
All tested concentrations were toxic to bacteria (strain specific) and could therefore only partly be used for evaluation up to and including 1800 µg per plate. All tests were negative in the first experiment. In the second experiment TA 98 produced a positive result, however this result was not dose dependent or repeatable and therefore deemed to be a random occurrence. The substance started to precipitate at 450 µg per plate. The positive controls increased the mutant counts to over double those of the negative controls.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
HF: Mammalian cells (cytogenicity) study (pre-OECD): Positive clasogenicity in bone marrow, negative dominant lethal (germ cells) (Veroshilin et al, 1975)
HF: Mammalian cells (mutation) studies (pre-OECD): Positive (Gerdes, 1971 and Mohamed, 1971).
NaF (read-across analogue): Mammalian cells (cytogenicity): Negative cytogenicity in bone marrow (Zeiger et al, 1994).
Link to relevant study records
- Endpoint:
- in vivo mammalian germ cell study: cytogenicity / chromosome aberration
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1994
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Published study
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- The study was performed in the mouse and investigated the incidence of chromosomal aberrations in bone marrow cells and the incidence of micronuclei in erythrocytes.
- GLP compliance:
- not specified
- Type of assay:
- other: combined chromosomal aberration and micronucleus assay
- Species:
- mouse
- Strain:
- not specified
- Sex:
- not specified
- Details on test animals or test system and environmental conditions:
- No further information given.
- Route of administration:
- oral: drinking water
- Vehicle:
- The test substance was administered in the water.
- Details on exposure:
- The cytogenetic effects of sodium fluoride (NaF) were measured in mice following administration in the drinking water for 6 weeks.
- Duration of treatment / exposure:
- Sodium fluoride was administered in the drinking water for 6 weeks.
- Frequency of treatment:
- Sodium fluoride, administered in the drinking water was available ad libitum for 6 weeks.
- Post exposure period:
- No post exposure period.
- Remarks:
- Doses / Concentrations:
200, 400 mg/L
Basis:
nominal in water - Control animals:
- yes, concurrent no treatment
- Positive control(s):
- A concurrent postive control was used.
- Tissues and cell types examined:
- Micronuclei were measured in peripheral blood erythrocytes following 1 and 6 weeks of administration. Bone marrow cell preparations were examined for the presence of chromosome aberrations following 6 weeks of treatment.
- Details of tissue and slide preparation:
- Micronuclei were measured in peripheral blood erythrocytes following 1 and 6 weeks of NaF administration. Bone marrow cell preparations were examined for the presence of chromosome aberrations following 6 weeks of treatment; metaphase and anaphase cells were examined. Anaphase cells were scored in three independent laboratories, two of which also scored metaphase cells from the same slides.
- Evaluation criteria:
- No evaluation criteria.
- Statistics:
- Not relevant
- Sex:
- male/female
- Genotoxicity:
- negative
- Remarks:
- No evidence of micronuclei in erythrocytes or cheomosomal aberrations in bone marrow cells.
- Toxicity:
- yes
- Remarks:
- Mortality at 400 mg/L; bodywieght effects at 200 and 400 mg/L
- Vehicle controls validity:
- valid
- Negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- Bone fluoride levels were determined and showed a dose-related incorporation of fluoride. No increases in micronuclei were seen in peripheral erythrocytes at either time point, and no increases in chromosome aberrations were seen in bone marrow cells when metaphase or anaphase cells were examined.
- Conclusions:
- Interpretation of results (migrated information): negative
Sodium fluoride did not cause any cytogenetic effects in mice under the conditions of this study - Executive summary:
The potential for sodium fluoride to cause chromosomal effects was investigated in a drinking-water study in mice. Mice were exposed for 1 or 6 weeks to sdoium fluoride. No evidence of micronuclei formation was seen in peripheral blood erythrocytes afer 1 or 6 weeks; no evidence of chromosomal aberration was seen in bone marrow cells after exposure for 6 weeks. Marked toxicity (including mortality) was seen at the highest dose level in this study.
- Endpoint:
- in vivo mammalian germ cell study: gene mutation
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 1971
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- other: Published non-guideline study.
- Principles of method if other than guideline:
- Drosophila were exposed to hydrofluoric acid vapour to determine the level of induced recessive lethals.
- GLP compliance:
- not specified
- Type of assay:
- other: Drosophila recessive lethal test
- Species:
- Drosophila melanogaster
- Strain:
- not specified
- Sex:
- male
- Details on test animals or test system and environmental conditions:
- 3 day old Drosophila melanogaster of the +b/dp+ genotype. Flies were allowed to acclimatise to the population cages for 15 minutes before the start of the experiment.
- Route of administration:
- inhalation: vapour
- Vehicle:
- Hydrofluoric acid was administered in a 2.5% aqueous solution. Controls were exposed to double-distilled water.
- Details on exposure:
- The males were exposed to hydrofluoric acid vapour in fumigation cages. Details of this method are reported in another paper that is unavailable. Exposures were conducted at room temperature. The first group of flies were removed after 6 hours exposure, the second group after 9 hours exposure, and the third group after 12 hours exposure. After each period the exposed males and concurrent controls were mated singly with virgin females of the tester stock. Controls were exposed to double-distilled water only.
- Duration of treatment / exposure:
- 6 hours, 9 hours and 12 hours.
- Frequency of treatment:
- Single exposure.
- Post exposure period:
- Two generations.
- Remarks:
- Doses / Concentrations:
2.5%
Basis:
nominal conc.
2.5% acid - No. of animals per sex per dose:
- No information.
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- Not examined.
- Tissues and cell types examined:
- Not applicable.
- Details of tissue and slide preparation:
- Not applicable.
- Evaluation criteria:
- Relative viabilities of homozygous chromosome 2, based on an expected value of 33.3% +b or +dp because twice as many Pm b or Pm dp are expected as homozygous for chromosome 2.
- Statistics:
- T-tests.
- Sex:
- male/female
- Genotoxicity:
- positive
- Toxicity:
- not examined
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- not examined
- Additional information on results:
- Hydrofluoric acid exposure reduced the viability of homozygotes for all 3 exposure periods.
- Conclusions:
- Interpretation of results (migrated information): ambiguous
Exposure to hydrofluoric acid vapour reduced the viability of homozygotes, however this test does not comply with guidelines and therefore it is difficult to reach conclusions regarding the results. - Executive summary:
Male Drosophila melanogaster of the genotype +b/dp+ were exposed to 2.5% hydrofluoric acid vapour (or double-distilled water) for 6, 9 or 12 hours. They were then crossed individually to virgin females of the genotype Cy/Pm dp b. Data from the second generation showed that hydrogen fluoride reduced the viability of homozygotes for the second chromosome, and this effect tended to increase with increasing exposure time. The authors concluded that the results were indicative of a cumulative effect of hydrofluoric acid exposure and that exposure caused gene mutations that affect viability. However it should be noted that the study is not guideline compliant, therefore it is difficult to reach conclusions regarding the genotoxiciy of hydrogen fluoride in this instance.
- Endpoint:
- in vivo mammalian germ cell study: gene mutation
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 1971
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- other: Published study, poorly reported.
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- Exposure to 3 concentrations of hydrogen fluoride vapour to determine sex-linked recessive lethal mutations and sterility levels in Drosophila melanogaster.
- GLP compliance:
- no
- Remarks:
- : older published study, pre-dates GLP
- Type of assay:
- Drosophila SLRL assay
- Species:
- Drosophila melanogaster
- Strain:
- other: Oregon-r
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- Oregon-r stocks of Drosophila melanogaster adults (8-12 hours old). Populations were maintained at stable densities by removing in sequence the food vials. Virgin "Basc" females were used to find sex-linked lethals.
- Route of administration:
- inhalation: vapour
- Vehicle:
- No vehicle was used.
- Details on exposure:
- Drosophila were exposed to hydrogen fluoride vapour in a fumigation chamber.
- Duration of treatment / exposure:
- 24 hours for the sex-linked recessive lethal test. Sterility testing was conducted after 0, 3 and 6 weeks exposure.
- Frequency of treatment:
- Continuous during exposure period.
- Post exposure period:
- Matings were carried out over three 3-day brood periods. Egg samples were collected at 0, 3 and 6 weeks from initiation of fumigation.
- Remarks:
- Doses / Concentrations:
0, 1.3, 2.9 and 4.3 ppm
Basis:
nominal conc.
vapour - No. of animals per sex per dose:
- No information.
- Control animals:
- yes, concurrent no treatment
- Positive control(s):
- Not examined.
- Tissues and cell types examined:
- Not applicable.
- Details of tissue and slide preparation:
- Not applicable.
- Evaluation criteria:
- Sex-linked recessive lethals: The F1 generation from the treated males and Basc females were allowed to intermate and the females then sorted singly into vials. The absence of wild type males in the F2 generation was scored as a lethal.
Sterility: Each treated male and female was mated individually to a tester and these crosses were scored after 5 days for the presence of larvae. If no larvae were present the treated parent was transferred to a new vial and remated to a tester. After an additional 5 day period the flies in the vials without larvae were scored as sterile. - Statistics:
- Lethal frequencies: Pearson and Hartley test for significance of observed differences between two Poisson variables.
Sterility: expressed as the percentage failure of adult Drosophila to produce viable larvae, ANOVA was used to determine significance of differences. - Sex:
- male/female
- Genotoxicity:
- positive
- Remarks:
- 2.9 ppm and above
- Toxicity:
- not examined
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Additional information on results:
- No additional information.
- Conclusions:
- Interpretation of results (migrated information): ambiguous
Sub-lethal exposures to hydrogen fluoride vapour resulted in genetic aberrancies in Drosophila melanogaster, however the study was poorly reported therefore the significance of the results cannot be evaluated. - Executive summary:
Drosophila melanogaster were exposed to hydrogen fluoride vapour to investigate levels of mutagenicity caused by the substance. Significant increases in mutation frequency were observed at 2.9 and 4.3 ppm concentrations (24 hour exposure). The percentage sterility was increased for both males and females, sterility increased with time exposed (0, 3 and 6 weeks) and with concentration. The author concluded that sex-linked recessive lethals indicated an effect upon a specific chromosome, whilst the increase in sterility levels resulted from a broad effect upon the genome as a whole. However it must be noted that the study was poorly reported and therefore the significance of these results cannot be evaluated.
- Endpoint:
- in vivo mammalian germ cell study: cytogenicity / chromosome aberration
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 1975
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- other: English abstract of a Russian study; poor description of methodology and results
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- Chromosomal aberration in rat bone marrow cells following inhalation of HF; dominant lethal assay in mice following HF inhalation
- GLP compliance:
- no
- Remarks:
- : older publsihed study, pre-dates GLP
- Type of assay:
- chromosome aberration assay
- Species:
- other: Rat and mouse
- Route of administration:
- inhalation
- Remarks:
- Doses / Concentrations:
1 mg/m3
Basis:
nominal conc. - Tissues and cell types examined:
- Bone marrow (rat); dominant lethal effect (mouse)
- Genotoxicity:
- positive
- Remarks:
- : rat bone marrow clastogenicity
- Genotoxicity:
- negative
- Remarks:
- : mouse dominant lethal
- Conclusions:
- Interpretation of results (migrated information): other: unreliable
The authors report clastogenicity in the bone marrow cells of rats exposed to HF by inhalation, but no dominant lethal effect in mice. However the study is not considered to be reliable. - Executive summary:
Rats and mice were exposed by inhalation to a nominal concentration of 1.0 mg/m3 HF. Bone marrow chromosomal aberrations were investigated in rats; a dominant lethal effect was investigated in mice.
The authors report clastogenicity in the bone marrow cells of rats exposed to HF by inhalation, but no dominant lethal effect in mice. However the study is not considered to be reliable due to poor description of methods and results.
Referenceopen allclose all
Bone fluoride levels were determined and showed a dose-related incorporation of fluoride. No increases in micronuclei were seen in peripheral erythrocytes at either time point, and no increases in chromosome aberrations were seen in bone marrow cells when metaphase or anaphase cells were examined. A concurrent positive control, cyclophosphamide, produced significant increases in peripheral blood cell micronuclei and in chromosome aberrations in bone marrow cells in metaphase. No increases in aberrations were seen in the same cyclophosphamide-treated mice when anaphase cells were examined.
The average viability for the tested 203 chromosomes was 26.26±1.24%. T-tests indicated that the mean difference of 0.71% between 6 and 9 hour exposure times was highly significant, and the same was true between the 9 and 12 hour exposure durations.
The lethal frequency results of the three 3 -day broods were pooled. 1.3ppm hydrogen fluoride caused a non-significant increase in mutation frequency (0.015%) compared to controls. Significant increases in mutation frequency were observed at 2.9 and 4.3ppm concentrations. The percentage sterility was increased for both males and females, sterility increased with time and with concentration.
Prolonged inhalation of this compound increased the frequency of cells with chromosome abnormalities in the bone marrow of albino rats; the effect was greater in older animals. No dominant-lethal effect was observed in mice under the same conditions.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Genetic toxicity in vitro
No evidence of mutagenicity was seen in a guideline-compliant GLP Ames test (Herbold, 1987). No evidence of mutagenicity was seen with sodium fluoride in an Ames test (NTP, 1990). No evidence of mutagenicity was seen in a mammalian cell mutation assay (V79/HPRT) with sodium fluoride. This study was performed only in the absence of metabolic activation, however this deviation is not considered to be critical as the test substance is not metabolised. A positive result with sodium fluoride is reported in a mouse lynmphoma assay (NTP, 1990). Sister chromatid exchange and chromosomal aberrations are reported in an additonal NTP study.
Genetic toxicity in vivo
Gerdes (1971) reports a marginally (but not statistically significant) positive response in a study in Drosophila melanogaster; positive effects in Drosophila are also reported by Mohamed et al (1971). The significance of these results is unclear; the EU RAR for HF considers the findings of these two Drosophila studies to be inconclusive. Zeiger et al (1994) report no evidence of clastogenicity, even at dose levels causing severe toxicity, in a well-conducted mouse study performed with sodium fluoride in which chromosomal aberrations and micronucleus formation was assessed. In contrast, a poorly reported inhalation exposure study performed with HF (Voroshilin et al, 1975) reports clastogenicity in the bone marrow of exposed rats but no dominant lethal effect in exposed mice.
Conclusion
The EU RAR concludes that, while the dataset on the genotoxicity of HF is limited, studies with sodium fluoride are also informative as for both substances target tissues will exposed to fluoride (either free or bound to organic molecules). The EU RAR therefore reviews the available data for NaF and HF and concludes that fluoride does not interact directly with DNA and is not genotoxic when administered via an appropriate route (i.e. by oral or inhalation exposure).
Short description of key information:
Studies with HF and NaF are available.
Endpoint Conclusion: No adverse effect observed (negative)
Justification for classification or non-classification
No classification is proposed. The available data indicate that fluoride does not interact directly with DNA and is not genotoxic when administered via an appropriate route (i.e. by oral or inhalation exposure).
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