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EC number: 242-354-0 | CAS number: 18472-51-0
- 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
Link to relevant study records
- 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:
- 2001-12-12 to 2002-03-26
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix (Aroclor 1254-induced rat liver)
- Test concentrations with justification for top dose:
- Plate incorporation test / Preincubation test: 0, 0.1, 0.316, 1, 3.16 and 10 ug/plate (concentrations are referred to the active ingredient).
The top concentration was chosen based on a preliminary test in TA 100 with 10 concentrations ranging from 0.316 - 5000 ug/plate, where pronounced to complete cytotoxicity was observed at >= 10 µg/plate. - Vehicle / solvent:
- The test substance was dissolved in water and added to the incubation assay.
- Untreated negative controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: without metabolic activation: sodium azide (TA 1535, TA 100); 2-nitrofluorene (TA 98); 9-aminoacridine (TA 1537); methylmethanesulfonate (TA 102); with metabolic activation: cyclophosphamide (TA 100, TA 1535); 2-anthraceneamide (TA 98, TA 102, TA 1537)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: plate incorporation and preincubation
Preincubation period: Plate incorporation test: none; Preincubation test: 60 min at 37 °C
Plates: 3 per concentration; 2 independent experiments - Evaluation criteria:
- Number of revertants is significantly increased compared to solvent controls to at least 2-fold (TA 98, TA 100, TA 102) and 3-fold (TA 1535, TA 1537) in both independent experiments
- Statistics:
- U-test according to Mann & Whitney
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- plate incorporation test: without S9 mix: at 10 ug/plate; with S9 mix: no cytotoxicity; preincubation test: without S9 mix: all strains at >= 3.16 ug/plate; with S9 mix: at 10 ug/plate
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- plate incorporation test: without S9 mix: at 10 ug/plate; with S9 mix: no cytotoxicity; preincubation test: without S9 mix: >= 3.16 ug/plate
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- plate incorporation test: without S9 mix: at 10 ug/plate; with S9 mix: no cytotoxicity; preincubation test: without S9 mix: >= 3.16 ug/plate; with S9 mix: at 10 ug/plate
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- plate incorporation test: without S9 mix: at 10 ug/plate; with S9 mix: no cytotoxicity; preincubation test: without S9 mix: >= 3.16 ug/plate; with S9 mix: at 10 ug/plate
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 102
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- plate incorporation test: without S9 mix: at >= 3.16 ug/plate; with S9 mix: no cytotoxicity; preincubation test: without S9 mix: at >= 3.16 ug/plate; with S9 mix: at 10 ug/plate
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results: negative
No evidence for mutagenic activity in the Salmonella typhimurium assay. - Executive summary:
Mutagenicity study (reverse mutation assay) in Salmonella typhimurium according to OECD guideline 471 with plate incorporation test and preincubation test each with and without metabolic activation by S9 mix from Arochlor 1254-induced rat liver. The concentration of the test substance was based on the observations of a preliminary test with strain TA 100 in which pronounced to complete cytotoxicity was observed at >= 10 µg/plate. Under the conditions of the assay where positive controls exerted the expected mutagenic effects, chlorhexidine digluconate caused no mutagenic effect in the Salmonella typhimurium strains TA 98, TA 100, TA 102, TA 1535 and TA 1537 neither in the plate incorporation nor in the preincubation test each carried out with and without metabolic activation.
- 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:
- 2005
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Remarks:
- No explicit positive control was run with the test. However, several of the different test substances gave clearly positive results in the absence and presence of an exogenous metabolic activation indicating the validity of the assay under the performed conditions. The number of metaphases scored was only 100. However, there was a clear lack of a cytogenetic effect of chlorhexidine digluconate although the cells were continuously treated for a time equivalent of 1.5 normal cell cycle lengths. Therefore, the study is considered as comparable with the corresponding OECD guideline and the results of the study are considered relevant for the assessment of in vitro cytogenicity of chlorhexidine digluconate.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- see above
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- other: Syrian hamster embryo cells (SHE)
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix (rat, liver, induced with sodium phenobarbital and benzoflavone)
- Test concentrations with justification for top dose:
- 0, 0.3, 1, 3 and 10 µM
- Vehicle / solvent:
- The final test concentrations were obtained by first diluting with deionized water to 10 mg/mL and further with culture medium to the desired concentrations.
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- other: No explicit positive control was run with the test. However, several of the different test substances gave clearly positive results indicating the validity of the assay under the performed conditions
- Details on test system and experimental conditions:
- Tertiary cultures of SHE cells were treated with the test chemical for 24 h, i.e. 1.5 times the doubling time of the cells (16 h). In the presence of exogenous metabolic activation system, cells were treated with the test substance for 3 h followed by 18 h incubation in fresh medium. Cultures were treated 3 h before harvest with colcemide. Cytotoxicity was determined as colony-forming efficiency (relative to control) relative after 24 h of treatment with test substance and subsequent incubation for 7 days for colony formation.
- Evaluation criteria:
- For determination of chromosome aberrations, 100 metaphases were scored per experimental group. Achromatic lesions greater than the width of the chromatid were scored as gaps unless there was displacement of the broken piece of chromatid. If there was displacement, they were scored as breaks.
- Species / strain:
- other: Syrian hamster embryo cells (SHE)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 10 µM (-S9 mix)
- Vehicle controls validity:
- valid
- Conclusions:
- Interpretation of results: negative
No induction of chromosomal aberrations in a cytogenicity assay with SHE cells in the presence or absence of metabolic activation. - Executive summary:
Cytogenicity study in mammalian cells (Syrian hamster embryo cells, SHE) comparable to OECD guideline 473 with and without metabolic activation by S9 mix from phenobarbital/benzoflavone-induced rat liver. The test substance was applied up to the cytotoxic concentration range. Under the conditions of the assay where a number of other, simultaneously tested substances gave a positive response, chlorhexidine digluconate tested up to cytotoxic concentration of 10 µM did not induce chromosomal aberrations in a cytogenicity assay with SHE cells in the presence or absence of metabolic activation.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2001-12-12 to 2002-06-19
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- 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)
- Type of assay:
- mammalian cell gene mutation assay
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix (Aroclor 1254-induced rat liver)
- Test concentrations with justification for top dose:
- 1st trial: 0, 0.1, 0.5, 1, 5 and 10 ug/ml; 2nd trial: 1st trial: 0, 0.5, 1, 5, 10 and 20 µg/mL.
The top concentrations were chosen based on a preliminary cytotoxicity experiment, where pronounced cytotoxicity was observed at >= 10 µg/mL (+/- S9 mix). - Vehicle / solvent:
- The final test concentrations were obtained by dilution with culture medium which was then added to the cultures.
- Untreated negative controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: -S9 mix: ethylmethanesulphonate; +S9 mix: 9,10-dimethylbenzanthracene
- Details on test system and experimental conditions:
- Duration of exposure: 24 hrs without S9 mix and 4 hrs with S9 mix
- Evaluation criteria:
- If the test substance in both experiments does not increase the mutation frequency 2-fold above the mean of the solvent controls under any condition, or if the mutation frequency is always < 20 * 10E-6 and if at least 1,000,000 cells per condition have been evaluated, the compound is considered as negative in the test.
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- +/- S9 mix at >= 10 µg/mL
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results: negative
No evidence for mutagenic activity with and without metabolic activation. - Executive summary:
Mutagenicity study in mammalian cells (V79) according to OECD guideline 476 with and without metabolic activation by S9 mix from Arochlor 1254-induced rat liver. The concentration of the test substance was based on the observations of a pretest in which cytotoxicity was observed from >= 10 µg/mL. Under the conditions of the assay where positive controls exerted the expected mutagenic effects, chlorhexidine digluconate tested up to cytotoxic concentrations of 10 or 20 µg/mL did not induce forward mutations in the HPRT-V79 -mutagenicity assay in the presence or absence of metabolic activation.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1983
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Remarks:
- No official statement concerning GLP status, but study was controlled and signed by responsible study director. Only male mice were used. Cells from bone marrow were collected only once, at 6 h after the final treatment. Although there was no mortality at any dose, results were only reported for 6/10 treatment group. However, the presented results were consistent and clearly negative throughout. The positive control gave a positive response indicating the principle validity of the assay conditions. In summary, the results of the study are considered relevant as supportive data for the assessment of the genotoxicity of chlorhexidine digluconate.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- GLP compliance:
- no
- Remarks:
- No official statement concerning GLP status, but study was controlled and signed by responsible study director
- Type of assay:
- micronucleus assay
- Species:
- mouse
- Strain:
- Swiss
- Sex:
- male
- Details on test animals or test system and environmental conditions:
- Source: C.N.R.S, Orleans la Source
Age at study initiation: no data
Weight at study initiation: 30 g - Route of administration:
- intraperitoneal
- Vehicle:
- Vehicle: DMSO/glycerol (1+4 v/v)
Concentration in vehicle: no data - Details on exposure:
- Total volume applied: 10 or 20 mL/kg bw
- Duration of treatment / exposure:
- Number of applications: 2
Interval between applications: 24 h - Post exposure period:
- 6 h
- Remarks:
- Doses / Concentrations:
2 * 10, 2 * 20 or 2 * 30 mg/kg bw
Basis: - No. of animals per sex per dose:
- at least 5 m per dose and sampling time
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- Substance used as Positive Control: 2 * 2 mg/kg bw Mitomycin C (vehicle: water)
- Tissues and cell types examined:
- Tissue: bone marrow
- Details of tissue and slide preparation:
- Maximum tolerable dose: 30 mg/kg bw
Number of animals: all animals
Number of cells: 2000
Time points: 6 h after last treatment
Type of cells: erythrocytes in bone marrow - Evaluation criteria:
- Parameters: numbers and types of structural aberrations and ratio polychromatic/normochromatic erythrocytes
- Statistics:
- not further specified
- Sex:
- male
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- pilorection, dyspnoea, ataxie, pain at injection site
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results: negative
No increase of micronuclei in bone marrow cells at any dose above vehicle control. The positive control (Mitomycin) gave the expected positive response. - Executive summary:
Micronucleus test with male Swiss mice with intraperitoneal administration of chlorhexidine digluconate using three different dose levels and two treatments at 24 h interval. Assessment of micronuclei in bone marrow 6 h after the second treatment. Under the conditions of the assay, chlorhexidine digluconatedid not cause an increase in the frequency of micronuclei in bone marrow cells of male mice.
Reference
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
The results of in vitro genotoxicity studies with chlorhexidine digluconate in animals are summarised in the following Table.
Genotoxicity in vitro of chlorhexidine digluconate:
Test system |
Organism/ |
Concentrations tested |
Result |
Remark |
|
- S9 |
+ S9 |
||||
Ames test, reverse mutation in prokaryotes, OECD guideline 471 |
Salmonella typhimuriumTA98, TA100, TA102, TA1535, TA1537 |
Plate incorporation assay: 0.1-10 µgchlorhexidine digluconate/plate Preincubation assay: 0.1-10 µgchlorhexidine digluconate/plate |
- |
-a |
Guideline study Cytotoxicity: Plate incorporation assay at≥ 3.16 µg/plate (-S9) Preincubation assay: at≥ 3.16 µg/plate (‑ S9), at 10 µg/plate (+S9) |
Chromosomal aberrations, in-house method |
Syrian hamster embryo cells (SHE) |
0.3-10 µM |
- |
-b |
Comparable to guideline study Cytotoxicity at 10 µM (‑S9) |
Sister chromatid exchange, in-house method |
Syrian hamster embryo cells (SHE) |
0.3-10 µM |
- |
n.p.c |
Cytotoxicity at 10 µM |
HPRT test, forward point mutation in mammalian cells, OECD guideline 476 |
V79 Chinese hamster lung fibroblasts |
0.1-20 µ/mlchlorhexidine digluconate |
- |
- |
Guideline study Cytotoxicity at≥ 10 µg/ml (+ and ‑S9) |
a: Activation system S9 fraction from Arochlor 1254-induced rat
liver;
b: Activation system S9 fraction from Phenobarbital/benzoflavone-induced
rat liver
c: not performed
Chlorhexidine digluconate was not mutagenic in a Salmonella typhimurium preincubation and a plate incorporation assay performed according to OECD guideline 471.
In a cytogenetic study with Syrian hamster embryo (SHE) cells in vitro, chlorhexidine digluconate did not reveal any cytogenetic activity (induction of chromosomal aberrations) when tested up to cytotoxic concentrations. There was no explicit positive control substance in the test, but several of the different substances tested in the same investigation gave clearly positive results in the absence and presence of exogenous metabolic activation indicating the validity of the assay under the performed conditions. The number of metaphases scored was lower than recommended in the OECD guideline 473. However, chlorhexidine digluconate clearly lacked a cytogenetic activity even though the cells were continuously treated for a time equivalent of 1.5 normal cell cycle lengths. It is concluded that the study is valid and comparable with the corresponding OECD guideline.
Additionally, chlorhexidine digluconate did not induce sister chromatid exchange (SCE) in SHE cells (tested only in the absence of exogenous metabolic activation) in a further investigation of the same study group.
In a HPRT-mutagenicity study in vitro according to OECD guideline 474, chlorhexidine digluconate did not induce forward mutations when tested up to cytotoxic concentrations.
The results of in vivo genotoxicity studies in animals are summarised in the following Table.
Genotoxicity in vivo:
Type of test |
Species |
Frequency of application |
Sampling times |
Dose levels |
Results |
Micronucleus test |
Mouse, Swiss ≥ 5 males/group |
2 times, intraperitoneal, with 24 h interval |
6 h after last treatment |
2 x 10, 20, or 30 mg/kg in DMSO/ glycerol
|
Result at all dose levels: - No indication of a clastogenic effect in the bone marrow |
A micronucleus test was performed with male mice. The study provided no evidence of a clastogenic effect of chlorhexidine in the bone marrow.
Short description of key information:
Possible genotoxic/mutagenic effects of chlorhexidine digluconate
have been tested in several in vitro (e.g. Ames test, Chromosomal
aberrations, HPRT test) and in vivo systems (Micronucleus test) and in
all reliable and relevant tests negative results were obtained.
Endpoint Conclusion: No adverse effect observed (negative)
Justification for classification or non-classification
The results of several reliable in vitro and in vivo studies gave no indications for a possible genotoxic/mutagenic effect of chlorhexidine digluconate. Therefore, there is no need for a classification.
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