Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 231-442-4 | CAS number: 7553-56-2
- 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
Iodine and chemically related compounds have been tested for genotoxicity in a variety of test systems both in vitro and in vivo (See Additional information box, Table 1 below). Based on release of iodine ( I2) and similar mode of action, mutagenicity data from iodine, iodine tincture and PVP-I are considered interchangeable.
Therefore, based on the overall weight of evidence for in vitro genotoxicity, the conclusion for gene mutation in bacteria is equivocal, the conclusion for in vitro cytogenicity studies in mammalian cells is positive and the conclusion for in vitro gene mutation in mammalian cells is negative.
Link to relevant study records
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Published literature study, guideline-comparable with limitations: - No replicates reported - Lower number of metaphases scored than recommended by current OECD guideline. - No positive control.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- other: Syrian hamster embryo cells
- Metabolic activation:
- without
- Test concentrations with justification for top dose:
- 0, 400, 600 and 800 µM
- Vehicle / solvent:
- - Solvent used: DMSO 60 mg/mL
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- other: not required
- True negative controls:
- no
- Positive controls:
- no
- Remarks:
- Not reported
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: Overnight
- Exposure duration: 24 hours
- Expression time (cells in growth medium): Not reported
- Fixation time (start of exposure up to fixation or harvest of cells): Not reported
SPINDLE INHIBITOR (cytogenetic assays): Colcemid (GIBCO, Grand Island, NY, USA) 0.2 µg/mL
STAIN (for cytogenetic assays): Not reported
NUMBER OF REPLICATIONS: Not reported
NUMBER OF METAPHASES: 100 per experimental group
DETERMINATION OF CYTOTOXICITY
- Method: colony-forming efficiencies
Metaphase chromosomes were prepared as described by Tsutsui et al. (Mutat. Res. 373: 113-123, 1997) - Evaluation criteria:
- Statistical significancy between treated cell and control
- Statistics:
- P value, χ2 test
- Species / strain:
- other: Syrian hamster embryo cells
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not examined
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- The ability of iodine to induce chromosome aberrations was examined using Syrian hamster embryo (SHE) cells in a testing design similar to OECD testing guideline 473 . Under testing conditions, iodine induced chromosome aberrations in Syrian hamster embryo cells
- Executive summary:
The ability of iodine to induce chromosome aberrations was examined using Syrian hamster embryo (SHE) cells in a testing design similar to OECD testing guideline 473 . SHE cells in tertiary culture were plated, incubated overnight, and treated with iodine at varying concentrations for 24 H. Cells were harvested with 0.1% trypsin. Three hours before harvest, Colcemid was administered and metaphase chromosomes were prepared. 100 metaphases were scored per experimental group. Statiscally significant increase in the frequency of chromosome aberrations were induced in SHE cells treated at the two highest dose levels.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Published literature study, guideline-comparable with limitations: - No metabolic activated trial. - Lower number of metaphases were scored compared to current OECD guideline. - Based on a similar mode action, release of iodine (I2), data from the iodine, iodine tincture and PVP-I is considered interchangeable
- Justification for type of information:
- Based on a similar mode action, release of iodine (I2), data from the iodine, iodine tincture and PVP-I is considered interchangeable.
Further information is included as attachment to attached background material. - Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- other: Human dental pulp cells (D824 cells)
- Metabolic activation:
- without
- Test concentrations with justification for top dose:
- 3-h assay: 0.03%, 0.1%, 0.3%
30-h assay: 0.01%, 0.03%, 0.1% - Vehicle / solvent:
- No vehicle
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- Remarks:
- 3h treatment
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: In a first assay, cells were treated for 3h with the agent or 4-nitroquinoline 1-oxide (4NQO) as positive control. In a second assay cells were treated with iodine tincture for 30h.
- Expression time (cells in growth medium): In the first assay, cells were incubated for a further 27 h. In the second assay, cells were continously treated with iodine treated for 30 h, as indicated previously.
SPINDLE INHIBITOR (cytogenetic assays): Colcemid 0.02 µg/mL (GIBCO/Invitrogen, Tokyo, Japan), administered three hours before the end of the incubation period in both assays.
STAIN (for cytogenetic assays): Not reported
NUMBER OF REPLICATIONS: No data on replicates.
NUMBER OF METAPHASES EVALUATED: 100 metaphases per dose group (see result table for info on negative and positive controls).
DETERMINATION OF CYTOTOXICITY
- Method: relative cell number (number of cells treated relative to the number of cells in the control cultures x100). Briefly, D824 cells were plated in triplicate and incubated overnight. The cells were treated with iodine tincture at varying concentrations for 3 h. After washing with fresh medium, cells were incubated for further 27 h. The number of cells was counted after harvesting with 0.25% trypsin. For the 30h experiment, cells were treated for the same period of time.
OTHER EXAMINATIONS:
- Determination of polyploidy and endoreplication: Yes, but counted as chromosome aberrations.
Metaphase chromosomes were prepared as described by Tsutsui et al. (Mutat. Res. 373: 113-123, 1997) - Evaluation criteria:
- Statistical significance on the incidence of chromosomal aberrations between treated cultures and control.
- Statistics:
- P value, χ2 test
- Species / strain:
- mammalian cell line, other: D824 cells
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Remarks:
- 3 h exposure
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- mammalian cell line, other: D824 cells
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Remarks:
- At highest dose only, 30h exposure
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not examined
- Additional information on results:
- See tables below
- Conclusions:
- The ability of iodine tincture to induce chromosome aberrations in D284 cells was evaluated in a study desgin similar to OECD testing guideline 473. Iodine tincture induced chromosome aberrations (in a statistically significant manner) only at the highest dose after 30 h of treatment without metabolic activation in D284 cells.
- Executive summary:
The ability of iodine tincture to induce chromosome aberrations in D284 cells was evaluated in a study desgin similar to OECD testing guideline 473. In a first assay, D284 cells were treated for 3 h with the agent and then incubated for a further 27 h. In the second experiment, D284 cells were treated continously for 30h. In both cases, three hours before harvest, cells were treated with Colcemid and metaphase chromosome were prepared. Iodine tincture induced chromosome aberrations (in a statistically significant manner) only at the highest dose after 30 h of treatment without metabolic activation in D284 cells.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Published literature study, guideline-comparable with limitation: - Lower number of metaphases were scored compared to current OECD guideline. - Based on a similar mode action, release of iodine (I2), data from the iodine, iodine tincture and PVP-I is considered interchangeable
- Justification for type of information:
- Based on a similar mode action, release of iodine (I2), data from the iodine, iodine tincture and PVP-I is considered interchangeable.
Further information is included as attachment to attached background material. - Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- other: Human dental pulp cells (D824 cells)
- Metabolic activation:
- with and without
- Metabolic activation system:
- Rat liver postmitochondrial supernatant (PMS)
- Test concentrations with justification for top dose:
- 3-h treatment assay without metabolic activation: 0.03%, 0.1%, 0.3%
30-h treatment assay without metabolic activation: 0.03%, 0.1%, 0.3%
2-h treatment with metabolic activation: 0.03%, 0.1%, 0.3% - Vehicle / solvent:
- No vehicle
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- Remarks:
- Experiment 1: 3h treatment without PMS
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- no
- Remarks:
- Experiment 2: 30h treatment without PMS
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- yes
- Remarks:
- PMS control
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- Experiment 3: 2h treatment with PMS
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: In experiment 1, cells were treated for 3h with the agent or 4-nitroquinoline 1-oxide (4NQO) as positive control without metabolic activation. In experiment 2, cells were treated with povidone iodone for 30h without metabolic activation. In experiment 3, cells were treated for 2 hours with povidone iodine or cyclophosphamide (as positive control) and then incubated.
- Expression time (cells in growth medium): In experiment 1, cells were incubated for a further 27 h. In experiment 2, cells were continously treated with povidone iodine for 30 h. In experiment 3, cells were treated for 2 h with povidone iodine in the presence of 5% PMS or with 50 µM cyclophosphamide in the presence of PMS.
SPINDLE INHIBITOR (cytogenetic assays): Colcemid 0.02 µg/mL (GIBCO/Invitrogen, Tokyo, Japan), administered three hours before the end of the incubation period in both assays.
STAIN (for cytogenetic assays): Not reported
NUMBER OF REPLICATIONS: No data on replicates.
NUMBER OF METAPHASES EVALUATED: 100 metaphases per dose group (see result table for info on negative and positive controls).
DETERMINATION OF CYTOTOXICITY
- Method: relative cell number (number of cells treated relative to the number of cells in the control cultures x100). Briefly, D824 cells were plated in triplicate and incubated overnight. The cells were treated with povidone iodine at varying concentrations for 3 h. After washing with fresh medium, cells were incubated for a further 27 h. The number of cells was counted after harvesting with 0.25% trypsin. For the 30h experiment, cells were treated for the same period of time. To measure cytotoxicity of povidone iodine in the presence of 5% rat liver postmitochondrial supernatant (PMS) cells were treated with varying concentrations of the agent and PMS for 2h. After washings, cells were incubated for further 28h and the cells were counted.
OTHER EXAMINATIONS:
- Determination of polyploidy and endoreplication: Yes, but counted as chromosome aberrations.
OTHER: PMS was prepared from three male Sprague-Dawley rats pretreated by injection (intraperitoneal) with sodium phenobarbital at 30 mg/kg bw on day 0 and at 60 mg/kg bw at day 1, 2 and 3; 5,6-benzoflavone at 80 mg/kg bw was injected intraperitoneally on day 2. On day 4, livers were excised from the euthanised animals and were homogenized with a Brinkman Polytron in three volumes of 0.25 M sucrose at pH 7.4 containing 2mM MgCl2. and 20 mM HEPES. The PMS fraction was prepared by two successive centrifugations at 9000g for 10 min and 15000 g for 20 min. The supernatant was frozen and store in liquid nitrogen until use.
Metaphase chromosomes were prepared as described by Tsutsui et al. (Mutat. Res. 373: 113-123, 1997) - Evaluation criteria:
- Statistical significance of the difference in the incidences of chromosome aberrations between control cultures and cultures treated with povidone iodine.
- Statistics:
- P value, χ2 test
- Species / strain:
- mammalian cell line, other: D824 cells
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Remarks:
- 3 h exposure without PMS
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- mammalian cell line, other: D824 cells
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Remarks:
- 30 h exposure without PMS
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not examined
- Species / strain:
- mammalian cell line, other: D824 cells
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Remarks:
- 2 h exposure with PMS
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- See tables below
- Conclusions:
- The ability of povidone iodine to induce chromosome aberrations in D284 cells was evaluated in three different experiments in a study design similar to OECD testing guideline 473. Povidone iodine did not induce chromosome aberrations in any of the tested conditions.
- Executive summary:
The ability of povidone iodine to induce chromosome aberrations in D284 cells was evaluated in three different experiments in a study design similar to OECD testing guideline 473. In a first assay, D284 cells were treated for 3 h with povidone iodine without metabolic activation and then incubated for a further 27 h. In the second experiment, D284 cells were treated continously for 30h without metabolic activation. In a third experiment, D284 cells were treated with povidone iodine in the presence of 5% rat liver postmitochondrial supernatant for 2 h and then incubated for 28 h. In all cases, three hours before harvest, cells were treated with Colcemid and metaphase chromosomes were prepared. Povidone iodine did not induce chromosome aberrations with and without metabolic activation in D824 cells under testing conditions.
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Published literature study, compared to current OECD guideline presents deficiencies: - Limited number of S. typhimurium strains - No metabolic activation assay was included - Rationale for dose selection is not reported (no information on cytotoxicity) - Tests conditions differ markedly from current guideline in second experiment with S. thyphimurium - Results are expressed as inhibition zone in E. Coli experiment - No information on replicates. - Based on a similar mode action, release of iodine (I2), data from the iodine, iodine tincture and PVP-I is considered interchangeable
- Justification for type of information:
- Based on a similar mode action, release of iodine (I2), data from the iodine, iodine tincture and PVP-I is considered interchangeable.
Further information is included as attachment to attached background material. - Qualifier:
- according to guideline
- Guideline:
- other: B.N. Ames, in "Chemical Mutagens" vol 1, A. Hollaender, Ed. Plenum, New York, N.Y., 1971, pp 267-282
- Qualifier:
- according to guideline
- Guideline:
- other: E.E. Slater, M.D. Anderson, H.S. Rosenkranz. Cancer Res. 31, 970 (1971).
- GLP compliance:
- no
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium, other: TA 1530 and TA1538
- Species / strain / cell type:
- E. coli, other: E. Coli pol A+/A1-
- Metabolic activation:
- not specified
- Test concentrations with justification for top dose:
- Mutagenicity tests: 0.2, 2 and 20 µL of povidone-iodine solution (0.1 M in ascorbic acid) in S. typhimurium assays.
DNA modifying activity: 1, 2, 3.3, 5, 10 and 20 µL of povidone-iodine solution (0.1 M in ascorbic acid). - Vehicle / solvent:
- No vehicle used
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: Ethyl methanesulfonate, methyl methanesulfonate, β-propiolactone, 2-nitrofluorene
- Remarks:
- S. typhimurium experiments
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: Methyl methanesulfonate, β-propiolactone, Propyleneimine, chloramphenicol
- Remarks:
- E. Coli experiment
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: as impregnation on paper disk
DURATION
- Exposure duration: In first experiment, plates were incubated in the dark at 37 °C for 54 h. After that time, the number of mutants and the diameter of the zones of growth inhibition were measured. In a second experiment, bacteria were chilled to 4 °C, and povidone-iodine was added. At intervals, portions of the cells were removed, the povidone-iodine was decomposed by addition of ascorbic acid (final concentration 0.1 M), and 10-fold serial dilutions of the treated cells were spread onto the surface of agar for the determination of the number of revertants to histidine independence (mutants) as well as of the number of viable cells. Plates were incubated in the dark for 54 h prior to scoring. In the DNA Modifying activity experiment, test substance was impregnated on paper disks and deposited on top of the E. Coli layer. The cultures were incubated at 37ºC for 12 h and the diameter of the zones of inhibition were measured.
SELECTION AGENT (mutation assays): Histidine free medium
NUMBER OF REPLICATIONS: Not reported
OTHER: Medium for scoring mutants consisted on minimal medium supplemented with biotin and containing a trace of histidine. - Evaluation criteria:
- Number of mutants per plate and zone of inhibition
- Statistics:
- Not required
- Species / strain:
- S. typhimurium, other: TA1530 and TA1538
- Metabolic activation:
- not specified
- Genotoxicity:
- negative
- Remarks:
- incubated at 37 °C
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium, other: TA1530
- Metabolic activation:
- not specified
- Genotoxicity:
- positive
- Remarks:
- Mutagenicity was demonstrated only on strain TA1530 under special conditions (incubation at 4 °C)
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- not specified
- Genotoxicity:
- negative
- Remarks:
- Mutagenicity was demonstrated only on strain TA1530 under special conditions (incubation at 4 °C)
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Species / strain:
- E. coli, other: E. Coli pol A+/A1-
- Metabolic activation:
- not specified
- Genotoxicity:
- other: Povidone-iodine blocked the growth of DNA polymerase-deficient E. Coli
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Conclusions:
- Povidone-iodine was not mutagenic to strain TA1530 and TA1538 following the standard procedure developed by Ames. In the second experiment under no standard conditions, povidone showed mutagenic activity in strain TA1530 but not in TA1538.
In the E.Coli DNA Modifying activity assay, the diameter of zone of inhibition was increased in a dose dependent manner, but due to lack of cytotoxicity data, it is not possible to assess whether the effect observed is due to the bactericidal properties of the test agent.
The study is deemed ambiguous due to the lack of cytotoxicity data and the biological relevance of the no standard conditions where a positive result was observed with S. typhimurium. - Executive summary:
The mutagenic potential of povidone-iodine was assessed in two mutagenicity tests. In the first experiment, a standard Ames test using S. typhimurium TA1530 and TA1538 was performed. The test substance was incorporated onto the surface of agar plates using paper disks impregnated with the test agent. The plates were incubated for 54 h at 37 ℃. A second experiment was performed using bacteria in liquid medium chilled to 4 ℃ . At intervals, portions of cells were removed and povidone iodine was decomposed. The plates were in the dark for 54 h and the number of colonies were counted.
Additionally, a DNA modifying activity was conducted with E. Coli pol A+ and pol A1 -. In this case, paper disks were impregnated with the test substance and deposited on top of the bacterial layer. The cultures were incubated at 37 ℃ for 12 h.
Povidone-iodine was not mutagenic to strain TA1530 and TA1538 following the standard procedure developed by Ames. In the second experiment under no standard conditions, povidone showed mutagenic activity in strain TA1530 but not in TA1538.
In the E.Coli DNA Modifying activity assay, the diameter of zone of inhibition was increase in a dose dependent manner, but due to lack of cytotoxicity data, it is not possible to assess whether the effect observed is due to the bactericidal properties of the test agent.
The study is deemed ambiguous due to the lack of cytotoxicity data and the biological relevance of the testing conditions where a positive result was observed with S. typhimurium.
- Endpoint:
- in vitro DNA damage and/or repair study
- Remarks:
- Type of genotoxicity: DNA damage and/or repair
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Published literature study, guideline-comparable with limitations: - No replicates reported - No treatment with metabolic activation - No positive control - Culture conditions not reported
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 479 (Genetic Toxicology: In Vitro Sister Chromatid Exchange Assay in Mammalian Cells)
- GLP compliance:
- not specified
- Type of assay:
- sister chromatid exchange assay in mammalian cells
- Species / strain / cell type:
- mammalian cell line, other: Syrian hamster embryo cells
- Metabolic activation:
- without
- Test concentrations with justification for top dose:
- 0, 79, 236, 473 µM
- Vehicle / solvent:
- - Vehicle(s)/solvent(s): DMSO (final concentration 473 mM)
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- no
- Remarks:
- DMSO is widely accepted for genotoxicity tests
- True negative controls:
- no
- Positive controls:
- no
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 24 h in the presence of 5-bromodeoxyuridine (10 µg/mL) under dark conditions.
SPINDLE INHIBITOR: Colcemid (2 µg/mL), added 3 hours before harvest.
STAIN: Slides were stained for 15 min in a solution of Hoechst 33258 (50 µg/mL in water), washed, dipped in PBS(-) and exposed to UV (row of 6 F15TB/BLB fluorescent bulbs, 33 J/m2/s) for 1 hour at 55°C. The slides were then stained with 3% GIEMSA solution for 10 min.
NUMBER OF REPLICATIONS: Not reported
NUMBER OF CELLS EVALUATED: 30 per group
DETERMINATION OF CYTOTOXICITY
- Method: colony-forming efficiency. Colony forming efficiency was determined in SHE cells by treating them for 24 h at varying concentrations of iodine. After harvesting with trypsin, cells were replated in triplicateonto dishes at 2000 cells/dish and incubated for 7 days. The relative colony-forming efficiency was expressed as the number of colonies in the treated dishes divided by the number in the control dishes x100.
- Evaluation criteria:
- Statistical comparison to control values
- Statistics:
- P value, Student t test
- Species / strain:
- mammalian cell line, other: Syrian hamster embryo cells
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- not examined
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not examined
- Conclusions:
- The ability of iodine to induce sister-chromatid exchange were examined using Syrian hamster embryo cells in a testing design similar to OECD tsting guideline 479. Sister-chromatid exchanges were induced by iodine in Syrian hamster embryo cells under testing conditions
- Executive summary:
The ability of iodine to induce sister-chromatid exchange were examined using Syrian hamster embryo cells in a testing design similar to OECD tsting guideline 479. SHE cells were treated at varying concentrations of iodine for 24 h in the presence of 5 -bromodeoxyuridine under dark conditions. Sister chromatids were differentially stained and thirty second-division metaphases with diploid number of chromosomes were analyzed for sister-chromatid exchange frequency. Sister chromatid exchanges were induced by iodine (p < 0.01) in concentration-dependent manner.
- 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
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Published literature study, guideline-comparable with limitations: - No assay with metabolic activation was performed. - Number of concentrations tested lower than recommended by current OECD guideline. - Limited data on cytotoxicity results is reported. - No independent repeat to confirm negative result.
- 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
- Target gene:
- TK (+/-)
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- The L5178Y cell line used in our studies was kindly supplied by Food and Drug Administration, Washington, D.C.. The cells were maintained in logarithmic growth by dilution three times a week to 3E+5 cells/mL with Fisher's Medium for Leukemic Mice (FMp), supplemented with 10% horse serum and 1% 200 mM glutamine (FM10) (Grand Island Biological Company, Grand Island, N.Y.). The cells were grown on a rocker platform (Bellco, Vineland, NJ.).
The cell lines were maintained at 37℃ in a humidified CO2 incubator. - Metabolic activation:
- without
- Test concentrations with justification for top dose:
- 22, 43, 85, 170 and 340 µg/mL
- Vehicle / solvent:
- No vehicle
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- no
- Remarks:
- not required
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- A second positive control was also used: N-methyl-N-nitro-N-nitrosoguanidine
- Details on test system and experimental conditions:
- Mutagenicity was assessed using a modification of the technique developed by Clive and Spector (Mutat. Res. 31: 17-29, 1975).
METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 hours (37 ºC)
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): 10 days (37 ºC)
SELECTION AGENT (mutation assays): 5-bromo-2'-deoxy-uridine (BUdR)
NUMBER OF REPLICATIONS: 6
DETERMINATION OF CYTOTOXICITY
- Method: Cell viability (trypan blue exclusion dye and by measurement of the cellular time). Dose level which resulted in 50% cell viability was used as highest in mutagenicity test. - Evaluation criteria:
- The number of mutant colonies/1E+5 cells was designated as having a mutational frequency (MF) of one. Mutational values of each respective dose were compared to the control value. A mutational frequency of 2.5 or greater was considered a positive mutational event.
- Statistics:
- Not required (except for positive controls, which were compared by using P value, student's T-Test).
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- The mutagenic potential of Iodine (I2) was studied using the L5178Y mouse (TK +/-) lymphoma assay in a study design similar to OECD testing guideline 476. Iodine did not showed biologically significant mutagenic ability under testing conditions.
- Executive summary:
The mutagenic potential of Iodine (I2) was studied using the L5178Y mouse (TK +/-) lymphoma assay in a study design similar to OECD testing guideline 476. Cells were incubated with the test compound for four hours at 37°C. The cells were then washed and incubated for additional 48 h to allow the expression of induced mutations. The cells were plated in Fishers Medium with 20% horse serum and in the presence of BUdR. Colony formation was scored after 10 days of incubation at 37°C.
All concentrations of iodine were inactive. In conclusion, iodine did not exert a biologically significant mutagenic ability in the mouse (TK +/-) lymphoma assay in vitro under tested conditions.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Published literature study, guideline-comparable with limitations: - Number of concentrations tested lower than recommended by current OECD guideline. - Limited data on cytotoxicity results is reported. - No independent repeat to confirm negative result. - Based on a similar mode action, release of iodine (I2), data from the iodine, iodine tincture and PVP-I is considered interchangeable-
- Justification for type of information:
- Based on a similar mode action, release of iodine (I2), data from the iodine, iodine tincture and PVP-I is considered interchangeable.
Further information is included as attachment to attached background material. - 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
- Target gene:
- TK (+/-)
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- The L5178Y cell line used in our studies was kindly supplied by Food and Drug Administration, Washington, D.C.. The cells were maintained in logarithmic growth by dilution three times a week to 3E+5 cells/mL with Fisher's Medium for Leukemic Mice (FMp), supplemented with 10% horse serum and 1% 200 mM glutamine (FM10) (Grand Island Biological Company, Grand Island, N.Y.). The cells were grown on a rocker platform (Bellco, Vineland, NJ.).
The cell lines were maintained at 37℃ in a humidified CO2 incubator. - Metabolic activation:
- with and without
- Metabolic activation system:
- S9 fraction of liver microsomes
- Test concentrations with justification for top dose:
- Experiment 1 (No metabolic activation): 10 mg/mL, 5 mg/mL, 1 mg/mL, 500 µg/mL and 100 µg/mL
Experiment 2 (with metabolic activation): 10 mg/mL, 5 mg/mL and 1 mg/mL - Vehicle / solvent:
- No vehicle (dissolved in media)
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- no
- Remarks:
- not required
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- N-dimethylnitrosamine
- Remarks:
- A second positive control was also used: N-methyl-N-nitro-N-nitrosoguanidine
- Details on test system and experimental conditions:
- Mutagenicity was assessed using a modification of the technique developed by Clive and Spector (Mutat. Res. 31: 17-29, 1975).
METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 hours (37 ºC)
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): 10 days (37 ºC)
SELECTION AGENT (mutation assays): 5-bromo-2'-deoxy-uridine (BUdR)
NUMBER OF REPLICATIONS: 6
DETERMINATION OF CYTOTOXICITY
- Method: Cell viability (trypan blue exclusion dye and by measurement of the cellular time). Dose level which resulted in 50% cell viability was used as highest in mutagenicity test. - Evaluation criteria:
- The number of mutant colonies/1E+5 cells was designated as having a mutational frequency (MF) of one. Mutational values of each respective dose were compared to the control value. A Mutational frequency of 2.5 or greater was considered a positive mutational event.
- Statistics:
- Statistically significance using P value.
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Conclusions:
- The mutagenic potential of povidone-iodine (PVP-I) was studied using the L5178Y mouse (TK +/-) lymphoma assay in a study dsign similar to OECD testing guideline 476. Povidone-iodine (PVP-I) did not show biologically significant mutagenic ability under testing conditions.
- Executive summary:
The mutagenic potential of povidone-iodine (PVP-I) was studied using the L5178Y mouse (TK +/-) lymphoma assay in a study dsign similar to OECD testing guideline 476. Cells were incubated with the test compound for four hours at 37 °C in the presence and absence of metabolic activation. The cells were then washed and incubated for additional 48 h to allow the expression of induced mutations. The cells were plated in Fishers Medium with 20% horse serum and in the presence of BUdR. Colony formation was scored after 10 days of incubation at 37 °C.
All concentrations of PVP-I were inactive in the absence of S-9 microsomal activation system. In the presence of S-9 fraction, PVP-I had only marginal activity as mutagen.
It was concluded that PVP-I did not possess any biologically significant mutagenic ability in the mouse (TK +/-) lymphoma assay in vitro under tested conditions.
- Endpoint:
- in vitro DNA damage and/or repair study
- Remarks:
- Type of genotoxicity: DNA damage and/or repair
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Published literature study, guideline-comparable with limitation: - Less cultures than recommended by current OECD guideline. - No positive control. - No assay with metabolic activation. - No second independent repeat for confirmation of the result.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 482 (Genetic Toxicology: DNA Damage and Repair, Unscheduled DNA Synthesis in Mammalian Cells In Vitro)
- GLP compliance:
- not specified
- Type of assay:
- other: DNA damage and repair assay, unscheduled DNA synthesis in mammalian cells in vitro
- Species / strain / cell type:
- mammalian cell line, other: Syrian hamster embryo cells
- Metabolic activation:
- without
- Test concentrations with justification for top dose:
- 3, 30, 300 µg/mL
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- no
- Positive controls:
- no
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: One hour in culture media
- Expression time (cells in growth medium): Six hours in 1mM hydroxyurea culture medium containing 370 KBq/mL [3H] thymidine.
Cells were washed with phosphate buffered saline (PBS) at 4 °C and were treated with 5% trichloro-acetate (TCA) at 4°C. 5% TCA was replaced 2-3 times at 1-2 hour interval. At the end cells were washed and dried. The cover slips were individually put into a scintillation vial containing toluene scintillater. The amount of [3H] thymidine collected which was not soluble in acid was measured in a liquid scintillation counter.
NUMBER OF REPLICATIONS: 4 cultures.
DETERMINATION OF CYTOTOXICITY
- Method: Cell survival rates.
Cells were put into solutions with a predetermined concentration of the test substance and left to react for 48 h. The cells were then washed with PBS twice and treated with trypsin to make a solution with cells. 2000 cells were plated onto four Petri dishes. After one week incubation, cells in the Petri dishes were washed with physiological saline and fixed with methyl alcohol before staining with Giemsa. Number of generated colonies were counted under a microscope. The cell survival rate was expressed as a percentage of a number of colonies in a treated group in comparison to the control group. - Evaluation criteria:
- Statistical difference from control
- Statistics:
- P values and Student t test
- Species / strain:
- mammalian cell line, other: Syrian hamster embryo (SHE) cells
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- not examined
- Untreated negative controls validity:
- not applicable
- True negative controls validity:
- not applicable
- Positive controls validity:
- not examined
- Conclusions:
- The ability of iodine to induce Unscheduled DNA synthesis (UDS) was examined in Syrian hamster embryo (SHE) cells in a study design simliar to OCD testing guidline 482. Iodine did not induce a statistically significant level of unscheduled DNA synthesis in SHE cells under testing conditions.
- Executive summary:
The ability of iodine to induce Unscheduled DNA synthesis (UDS) was examined in Syrian hamster embryo (SHE) cells in a study design simliar to OCD testing guidline 482. SHE cells were treated with iodine for 1h and then incubated for six in hydroxyurea culture medium containing [3H] thymidine. The uptake of [3H] thymidine was measured in a liquid scintillation counter. Iodine did not induce a statistically significant level of unscheduled DNA synthesis in SHE cells under testing conditions.
Referenceopen allclose all
Table 1: The ability of iodine to induce chromosome aberrations in SHE cells.
Conc. (µM) |
Relative colony forming efficiency (%) |
Type of aberrations (%) |
Aberrant metaphases (%) |
||||||||
CG |
ICG |
CB |
ICB |
E |
O |
D |
D |
F |
|||
0 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
400 |
75 |
2 |
3 |
1 |
1 |
0 |
0 |
0 |
0 |
0 |
5.0 |
600 |
62 |
9 |
5 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
13.0* |
800 |
61 |
10 |
8 |
1 |
2 |
2 |
0 |
0 |
0 |
0 |
19.0* |
CG, Chromatid gaps; ICG, isochromatid gaps; CB, chromatid breaks; ICB, isochromatid breaks; E, exchanges; O, ring chromosome; D, dicentric chromosomes; F, fragmentations. * Significantly different from control (P<0.01, χ2test).
Table 1: Ability of iodine tincture to induce chromosome aberrations in D824 cells treated for 3 h.
Conc. (%) |
Relative cell number (%) |
Number of metaphases scored (%) |
Type of aberrations (%) |
Aberrant metaphases (%) |
Polyploidy and endoreduplication |
|||||
G |
B |
E |
D |
O |
F |
|||||
0 |
100 |
500 |
1 |
0 |
0 |
0 |
0 |
0 |
1.0 |
3.6 |
0.03 |
70 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
4.0 |
0.1 |
68 |
100 |
1 |
0 |
0 |
0 |
0 |
0 |
1.0 |
7.0 |
0.3 |
60 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
3.0 |
4NQO(µM) |
|
|
|
|
|
|
|
|
|
|
0.5 |
59 |
100 |
7 |
11 |
1 |
0 |
0 |
0 |
19.0* |
5.0 |
1.0 |
52 |
100 |
9 |
21 |
1 |
0 |
0 |
0 |
25.0* |
19.0* |
G, gaps; B, breaks; E, exchanges; D, dicentric chromosomes; O, ring chromosomes; F, fragmentations; 4NQO, 4-nitroquinoline 1-oxide.
*Significantly different from control (P<0.01, χ2test)
Table 2: Ability of iodine tincture to induce chromosome aberrations in D824 cells treated for 30 h.
Conc. (%) |
Relative cell number (%) |
Number of metaphases scored (%) |
Type of aberrations (%) |
Aberrant metaphases (%) |
Polyploidy and endoreduplication |
|||||
G |
B |
E |
D |
O |
F |
|||||
0 |
100 |
300 |
1 |
0 |
0 |
0 |
0 |
0 |
1.0 |
3.0 |
0.01 |
110 |
100 |
1 |
0 |
0 |
0 |
0 |
0 |
1.0 |
1.0 |
0.03 |
64 |
100 |
1 |
0 |
0 |
0 |
0 |
0 |
1.0 |
1.0 |
0.1 |
63 |
100 |
5 |
0 |
0 |
0 |
0 |
0 |
5.0* |
0 |
G, gaps; B, breaks; E, exchanges; D, dicentric chromosomes; O, ring chromosomes; F, fragmentations; 4NQO, 4-nitroquinoline 1-oxide.
*Significantly different from control (P<0.05, χ2test)
Table 1: Ability of povidone-iodine tincture to induce chromosome aberrations in D824 cells treated for 3 h.
Conc. (%) |
Relative cell number (%) |
Number of metaphases scored (%) |
Type of aberrations (%) |
Aberrant metaphases (%) |
Polyploidy and endoreduplication |
|||||
G |
B |
E |
D |
O |
F |
|||||
0 |
100 |
500 |
1 |
0 |
0 |
0 |
0 |
0 |
1.0 |
3.6 |
0.03 |
90 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0.1 |
97 |
100 |
2 |
0 |
0 |
0 |
0 |
0 |
2.0 |
2.0 |
0.3 |
87 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
3.0 |
4NQO(µM) |
|
|
|
|
|
|
|
|
|
|
0.5 |
59 |
100 |
7 |
11 |
1 |
0 |
0 |
0 |
19.0* |
5.0 |
1.0 |
52 |
100 |
9 |
21 |
1 |
0 |
0 |
0 |
25.0* |
19.0* |
G, gaps; B, breaks; E, exchanges; D, dicentric chromosomes; O, ring chromosomes; F, fragmentations; 4NQO, 4-nitroquinoline 1-oxide.
*Significantly different from control (P<0.01, χ2test)
Table 2: Ability of povidone-iodine tincture to induce chromosome aberrations in D824 cells treated for 30 h.
Conc. (%) |
Relative cell number (%) |
Number of metaphases scored (%) |
Type of aberrations (%) |
Aberrant metaphases (%) |
Polyploidy and endoreduplication |
|||||
G |
B |
E |
D |
O |
F |
|||||
0 |
100 |
300 |
1 |
0 |
0 |
0 |
0 |
0 |
1.0 |
3.0 |
0.03 |
94 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
5.0 |
0.1 |
77 |
100 |
2 |
0 |
0 |
0 |
0 |
0 |
2.0 |
4.0 |
0.3 |
65 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1.0 |
G, gaps; B, breaks; E, exchanges; D, dicentric chromosomes; O, ring chromosomes; F, fragmentations.
Table 3: Ability of povidone-iodine tincture to induce chromosome aberrations in D824 cells treated for 2 h in the presence of rat liver PMS.
Conc. (%) |
PMS (%) |
Relative cell number (%) |
Number of metaphases scored (%) |
Type of aberrations (%) |
Aberrant metaphases (%) |
Polyploidy and endoreduplication |
|||||
G |
B |
E |
D |
O |
F |
||||||
Control |
0 |
100 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
2.0 |
PMS Control |
5 |
99 |
100 |
1 |
0 |
0 |
0 |
0 |
0 |
1.0 |
2.0 |
P-I 0.03 |
5 |
101 |
100 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
4.0 |
P-I 0.1 |
5 |
84 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
5.0 |
P-I 0.3 |
5 |
63 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
4.0 |
CP(µM) |
|
|
|
|
|
|
|
|
|
|
|
50 |
0 |
78 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
1.0 |
50 |
5 |
51 |
100 |
25 |
15 |
6 |
0 |
0 |
0 |
40.0* |
1.0 |
G, gaps; B, breaks; E, exchanges; D, dicentric chromosomes; O, ring chromosomes; F, fragmentations; CP, cyclophosphamide; PMS, postmitochondrial supernatant; P-I, povidone-iodine.
*Significantly different from control (P<0.01, χ2test)
Povidone-iodine when tested following the standard procedure developed by Ames, no mutagenic activity was observed (Table 1). The known mutagens (methyl methanesulfonate, β-propiolactone, ethyl methanesulfonate and 2 -nitrofluorene) were active in the assay.
Table 1: Mutagenicity of povidone-iodine using a standard S. typhimurium assay system.
Agent |
Amount (µL) |
TA 1530 |
TA 1538 |
||
Zone of inhibition (mm) |
Mutants per plate |
Zone of inhibition (mm) |
Mutants per plate |
||
Povidone-iodine |
0.2 |
0 |
17 |
0 |
13 |
|
2 |
11.3 |
16 |
11.5 |
14 |
|
20 |
31.0 |
22 |
32.3 |
16 |
Ethyl methanesulfonate |
10 |
0 |
8000 |
0 |
7 |
Methyl methanesulfonate |
10 |
22.2 |
530 |
24.2 |
14 |
β-propiolactone |
10 |
31.0 |
2652 |
25.9 |
13 |
2-nitrofluorene |
250 µg |
0 |
13 |
0 |
87 |
Water (control) |
10 |
0 |
17 |
0 |
12 |
When using the modified experiment, povidone-iodine resulted active in S. typhimurium TA1530 in a concentration and time of exposure manner. Povidone-iodine had no mutagenic effect on strain TA1538. Both results are not reported in the article in a tabular way.
In the E. Coli assay, povidone-iodine, as indicated by the authors, blocked preferentially the growth of the DNA polymerase-deficient (Table 2).
Table 2: Inhibition of DNA polymerase-deficient E. Coli.
Agent |
Amount (µL) |
Diameter of zone of inhibition, mm |
|
Pol A+ |
Pol A1- |
||
Povidone-iodine |
20 |
73.0 |
>86 |
|
10 |
50.6 |
81.4 |
|
5 |
26.5 |
31.0 |
|
3.3 |
19.2 |
25.6 |
|
2 |
16.6 |
18.9 |
|
1 |
14.2 |
15.7 |
Methyl methanesulfonate |
10 |
37.4 |
55.1 |
β-propiolactone |
10 |
36.5 |
58.7 |
Propyleneimine |
10 |
11.5 |
21.5 |
Chloramphenicol |
30 µg |
31.8 |
31.9 |
Table 1: The ability of iodine to induce sister-chromatid exchange in SHE cells.
Conc. (µM) |
Relative colony forming efficiency (%) |
SCEs/cell |
|
0 |
100 |
8.03± 3.54 |
|
79 |
91 |
11.63 ± 4.60* |
|
236 |
90 |
12.67 ± 4.24* |
|
473 |
47 |
13.80 ± 3.81* |
* Significantly different from control (P<0.01, Studentttest) (n=30)
At doses tested, iodine did not induce significantly alter the mutation frequency in the mouse lymphoma assay (Table 1).
Table 1: Mutagenicity of iodine in the L5178Y test system.
Substance |
Concentration (µg/mL) |
Mutational Frequency (MF) |
Iodine |
340 |
0.76 |
|
170 |
1.09 |
|
85 |
2.0 |
|
43 |
0.84 |
|
22 |
0.67 |
Media |
- |
1.0* |
EMS |
500 |
12.0** |
MNNG |
5 |
15.0** |
EMS: Ethyl methanesulfonate; MNNG: N-methyl-N-nitro-N-nitrosoguanidine.
* Actual MF for media control before normalization to 1 was 4E-5mutants
** Statistical significance at levels of p<0.05
Povidone-iodine (PVP-I) at doses tested did not significantly alter the mutational frequency in the absence of metabolic activation (Table 1). In the same assay, both positive controls exhibited a significant mutagenic actional.
In the presence of metabolic activation, PVP-I produced a mutational frequency over 2.5, only at 5 mg/mL was the mutational frequency statistically significant from control values. The significance of the response with PVP-I at 10 mg/mL was difficult to assess due to high cytotoxicity (92% morbidity by trypan blue exclusion) was reported by the authors. Both positive controls (MNNG and DMN), were found to be potent mutagens. DMN was inactive in the absence of the S-9 fraction. The S-9 fraction alone had no effect on the mutational frequency.
Table 1: Mutagenicity of PVP-I in the L5178Y test system.
Substance |
Concentration |
Mutational Frequency (MF) |
PVP-I |
10 (mg/mL) |
2.33 |
|
5 (mg/mL) |
0.93 |
|
1 (mg/mL) |
0.75 |
|
500 (µg/mL) |
0.82 |
|
100 (µg/mL) |
0.75 |
Media |
- |
1.0* |
EMS |
500(µg/mL) |
12.0** |
MNNG |
5 (µg/mL) |
15.0** |
EMS: Ethyl methanesulfonate; MNNG: N-methyl-N-nitro-N-nitrosoguanidine.
* Actual MF for media control before normalization to 1 was 4E-5 mutants
** Statistical significance of levels of p<0.05
Table 2: Mutagenicity of PVP-I in the L5178Y test system with S-9 microsomal activation.
Substance |
Concentration |
Mutational Frequency (MF) |
PVP-I + S-95 |
10 (mg/mL) |
3.20 |
|
5 (mg/mL) |
4.741 |
|
1 (mg/mL) |
1.90 |
DMN |
3.70 (mg/mL) |
1.22 |
DMN + S-9 |
3.70 (mg/mL) |
252 |
MNNG |
5 (µg/mL) |
3.983 |
Media |
|
1.04 |
Media + S-9 |
|
1.04 |
DMN: Dimethylnitrosamine; MNNG: N-methyl-N-nitro-N-nitrosoguanidine; S-9: Aroclor 1254 activated rat liver microsomal homogenate.
1 Statistical significance at levels of p<0.01
2 Statistical significance at levels of p<0.01
3 Statistical significance at levels of p<0.001
4 MF for media control before normalization to 1 was 4 E-5 mutants
MF for media +S-9 before normalization was 6E-5mutants
520 µL of S-9 used per test sample
Table 1: Relative survival of SHE cells treated with iodine.
Concentration (µg/mL) |
Relative cell survival (%)* |
|
50 |
95.5 ± 10.7 |
|
100 |
91.3 ± 11.2 |
|
150 |
61.7 ± 7.1 ** |
* Mean ± S.D. of 4 dishes.
** Significantly different from control (P<0.01,ttest)
Table 2: Unscheduled DNA Synthesis (UDS) in SHE cells treated with iodine.
Concentration (µg/mL) |
Induced UDS* |
|
3 |
0 |
|
30 |
0 |
|
300 |
84.6 ± 9.9 |
* Mean ± S.D. of 4 cultures.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
Iodine and chemically related compounds have been tested for genotoxicity in a variety of systems both in vitro and in vivo (See Additonal information box, Table 1). Based on release of iodine (I2) and similar mode of action, mutagenicty data from the iodine, iodine tincture and PVP-I are considered interchangeable.
Therefore, based on the overall weight of evidence for in vivo genotoxicity, the conclusion for germ cell and somatic gene mutation is negative.
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Published literature study, guideline-comparable with limitations: - No positive control used - Sampling time was not at recommended by current OECD guideline - Lack of well documented rationale to confirm toxicity/availability in the target tissue - No data on mitotic index. - Based on a similar mode action, release of iodine (I2), data from the iodine, iodine tincture and PVP-I is considered interchangeable
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)
- GLP compliance:
- no
- Type of assay:
- mammalian bone marrow chromosome aberration test
- Species:
- hamster, Chinese
- Strain:
- other: Gricetulus griseus
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Thomae Co., Biberach, Federal Republic of Germany
- Age at study initiation: Not reported
- Weight at study initiation: Not reported
- Assigned to test groups randomly: Not reported
- Fasting period before study: Not reported
- Housing: Plastic (polycarbonate) cages
- Diet: Standardized feed (Altromin R supplied by Altromin Co. Lage/Lippe, Federal Republic of Germany or Ssniff H supplied by Intermast GmbH, Boeckum-Hoevel, Federal Republic of Germany), Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: Not reported
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 2
- Humidity (%): 55 ± 5
- Air changes (per hr): Not reported
- Photoperiod (hrs dark / hrs light): 12 h light/12 h dark - Route of administration:
- intraperitoneal
- Vehicle:
- - Solvent used: water
- Duration of treatment / exposure:
- Experiment 1: Single intraperitoneal administration (2 different doses: 38.3 or 82.5 mg/kg)
Experiment 2: Five intraperitoneal injections of one dose level 38.3 mg/kg - Frequency of treatment:
- In experiment 2, the intraperitoneal injections were administered in a 24-h interval
- Post exposure period:
- Experiment 1: Animals were sacrificed at 6, 24 and 48 after the single application
Experiment 2: Animals were sacrificed 6 hours after last application - Dose / conc.:
- 38.3 mg/kg bw/day (actual dose received)
- Remarks:
- Single or five applications
- Dose / conc.:
- 82.5 mg/kg bw/day (actual dose received)
- Remarks:
- Single application
- No. of animals per sex per dose:
- 6 male and 6 female per group
- Control animals:
- yes, concurrent no treatment
- yes, concurrent vehicle
- Positive control(s):
- No positive control was used
- Tissues and cell types examined:
- Metaphases from bone marrow
- Details of tissue and slide preparation:
- No further details reported
- Evaluation criteria:
- Statistical comparison to control
- Statistics:
- Significant differences were checked using chi-square or U test
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- repeated application caused the animals to show signs of pain lasting for ~1 min.
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Conclusions:
- Povidone-iodine USP was tested for genotoxic potential in Chinese Hamsters by the in vivo chromosome aberrations assay in the bone marrow in a study design similar to OECD testing guideline 475. Under testing conditions, povidone-iodine did not induce chromosome aberrations in the bone marrow of Chinese Hamster.
- Executive summary:
Povidone-iodine USP was tested for genotoxic potential in Chinese Hamsters by the in vivo chromosome aberrations assay in the bone marrow in a study design similar to OECD testing guideline 475.
To assess this potential, two experiments were carried out. In the first one, male and female hamsters were administered by single peritoneal injection with test compound, distilled water and a third group remained untreated. Animals were sacrificed after 6, 24 and 48 h post administration. In a second experiment, male and female hamsters were injected five times with a daily interval and sacrificed 6 hours after the last application. 100 metaphases per animal were examined for gaps, breaks and fragments.
There was no increase in the percentage of aberrant metaphases when the substance was administered singly or repeatedly. In conclusion, povidone-iodine did not induce chromosome aberrations in the bone marrow of Chinese Hamster, under testing conditions.
- Endpoint:
- in vivo mammalian germ cell study: cytogenicity / chromosome aberration
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Published literature study, guideline-comparable with limitations: - No positive control used nor historical data presented. - Only one dose level used with no detailed rationale on dose selection. - Mating schedule was not as recommended by current OECD guideline. - Based on a similar mode action, release of iodine (I2), data from the iodine, iodine tincture and PVP-I is considered interchangeable
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 478 (Genetic Toxicology: Rodent Dominant Lethal Test)
- GLP compliance:
- no
- Type of assay:
- rodent dominant lethal assay
- Species:
- mouse
- Strain:
- NMRI
- Sex:
- male
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Wiga Co., Sulzfeld, Federal Republic of Germany
- Age at study initiation: Not reported
- Weight at study initiation: Not reported
- Assigned to test groups randomly: Not reported
- Fasting period before study: Not reported
- Housing: Plastic (polycarbonate) cages
- Diet: Standardized fed (Altromin R supplied by Altromin Co. Lage/Lippe, Federal Republic of Germany or Ssniff H supplied by Intermast GmbH, Boeckum-Hoevel, Federal Republic of Germany), Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: Not reported
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 2
- Humidity (%): 55 ± 5
- Air changes (per hr): Not reported
- Photoperiod (hrs dark / hrs light): 12 h light/12 h dark - Route of administration:
- intraperitoneal
- Vehicle:
- - Solvent used: water
- Duration of treatment / exposure:
- Single intraperitoneal administration
- Frequency of treatment:
- Single intraperitoneal administration
- Post exposure period:
- Matings in week 1, 2, 3, 4, 5, 6, 7 and 8
- Dose / conc.:
- 72 mg/kg bw/day (actual dose received)
- No. of animals per sex per dose:
- 20 male mice
- Control animals:
- yes, concurrent no treatment
- yes, concurrent vehicle
- Positive control(s):
- No positive control used
- Tissues and cell types examined:
- Implantations (total), live embryos and dead implantations
- Details of tissue and slide preparation:
- No details reported
- Evaluation criteria:
- Mutagenicity index
- Statistics:
- Significant differences were checked using chi-square or U test
- Sex:
- male
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Conclusions:
- The potential genotoxicity of Povidone-iodine was evaluated in the Dominant Lethal Test in a study design similar to OECD testing guideline 478. Povidone-iodine was not genotoxic in the Dominant Lethal Assay under testing conditions.
- Executive summary:
The potential genotoxicity of Povidone-iodine was evaluated in the Dominant Lethal Test in a study design similar to OECD testing guideline 478. The assay is an indirect method for detecting effects on the germinal tissue of the tested species. Dominant lethals are generally recognized to be the result of chromosome aberrations, but gene mutations and toxic effects cannot be excluded.
In this assay, 20 male mice were administered 72 mg/kg povidone-iodine as a single peritoneal administration. Males were mated sequentially to three untreated females. After sacrifice of the females, the conception rate, total number of implantations, live embryos and dead implantations were counted.
The conception rate decreased significantly during the last week, but the average number of implantations and the mutagenicity index were not affected. During the other mating periods, all parameters varied in a similar manner as in the control animals.
It was concluded that povidone-iodine did not induce a genotoxic effect on mice in the dominant lethal test under described conditions.
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Published literature study, guideline-comparable with limitations: - No positive control used - Samples taken earlier than recommended by current OECD guideline - No repeat to confirm negative result - Lack of PCEs/NCEs ratio - Lack of well documented rationale to confirm toxicity/availability in the target tissue. - Based on a similar mode action, release of iodine (I2), data from the iodine, iodine tincture and PVP-I is considered interchangeable
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- GLP compliance:
- no
- Type of assay:
- mammalian erythrocyte micronucleus test
- Species:
- mouse
- Strain:
- NMRI
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Wiga Co., Sulzfeld, Federal Republic of Germany
- Age at study initiation: Not reported
- Weight at study initiation: Not reported
- Assigned to test groups randomly: Not reported
- Fasting period before study: Not reported
- Housing: Plastic (polycarbonate) cages
- Diet: Standardized fed (Altromin R supplied by Altromin Co. Lage/Lippe, Federal Republic of Germany or Ssniff H supplied by Intermast GmbH, Boeckum-Hoevel, Federal Republic of Germany), Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: Not reported
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 2
- Humidity (%): 55 ± 5
- Air changes (per hr): Not reported
- Photoperiod (hrs dark / hrs light): 12 h light/12 h dark - Route of administration:
- intraperitoneal
- Vehicle:
- - Solvent used: water
- Duration of treatment / exposure:
- Double intraperitoneal application
- Frequency of treatment:
- Double intraperitoneal application in a 24-h interval
- Post exposure period:
- 6 h after last dose, animals were sacrificed
- Dose / conc.:
- 36 mg/kg bw/day (actual dose received)
- No. of animals per sex per dose:
- 10 animals per group (five male and five female)
- Control animals:
- yes, concurrent no treatment
- yes, concurrent vehicle
- Positive control(s):
- No positive control used
- Tissues and cell types examined:
- Normochromatic (4000 per animal) and polychromatid (2000 per animal) erythrocytes from bone marrow (femurs) were examined
- Details of tissue and slide preparation:
- No further details on smear preparations
- Evaluation criteria:
- Statistical significance compared to control
- Statistics:
- Significant differences were checked using chi-square or U test
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Conclusions:
- Povidone-iodine was tested for genotoxicity in mice by the micronucleus test in a study design similar to OECD testing guideline 474. Povidone-iodine did not induce significant increase in frequency of micronucleated cells in bone marrow erythrocytes under testing conditions.
- Executive summary:
Povidone-iodine was tested for genotoxicity in mice by the micronucleus test in a study design similar to OECD testing guideline 474.
Three groups of 10 animals (5 male and 5 females) were used, one group received 36 mg povidone-iodine/kg (one-tenth LD50), another group received 10 mL of distilled water/kg twice at an interval of 24 h. The third group of animals remained untreated. Six hours after the final dose, the animals were sacrificed, the femurs were removed and bone marrow smears were prepared. Normochromatic and polychromatic erythrocytes were examined for the frequency of micronucleated cells. There was merely a slight increase in polychromatic erythrocytes containing micronuclei, but even this increase was within the normal range.
The findings indicated that povidone-iodine was not genotoxic by the micronucleus test in somatic cells under testing conditions.
Referenceopen allclose all
There was no increase in the percentage of aberrant metaphases when the substance was administered singly (Table 1) or repeatedly (Table 2). The animals tolerated povidone-iodine with no symptoms when administered in a single dose, but repeated application caused the animals show signs of pain lasting for approximately 1 min.
Table 1: Testing povidone-iodine in the bone marrow test on Chinese Hamster with single intraperitoneal injection
Dose |
Untreated |
10 mL of saline/kg |
38.3 mg/kg |
82.5 mg/kg |
||||||||
Put to death (h) |
6 |
24 |
48 |
6 |
24 |
48 |
6 |
24 |
48 |
6 |
24 |
48 |
No of animals |
12 |
11 |
12 |
12 |
12 |
12 |
12 |
11 |
12 |
12 |
12 |
12 |
Analyzed metaphases |
1200 |
1100 |
1200 |
1200 |
1200 |
1200 |
1200 |
1100 |
1200 |
1200 |
1200 |
1200 |
Animals with aberrant metaphases |
8 |
8 |
9 |
7 |
9 |
6 |
9 |
8 |
8 |
6 |
9 |
9 |
Metaphases with aberrations |
|
|
|
|
|
|
|
|
|
|
|
|
Including gaps |
16 |
12 |
18 |
11 |
22 |
11 |
18 |
23 |
15 |
12 |
20 |
21 |
Excluding gaps |
3 |
4 |
2 |
2 |
2 |
2 |
4 |
5 |
2 |
2 |
2 |
5 |
Gaps (chromatide) |
9 |
8 |
14 |
7 |
15 |
9 |
10 |
15 |
10 |
7 |
12 |
16 |
Gaps (isochromatide) |
4 |
0 |
2 |
2 |
5 |
0 |
4 |
3 |
3 |
3 |
6 |
0 |
Breaks (chromatide) |
0 |
2 |
1 |
1 |
0 |
2 |
2 |
2 |
2 |
1 |
0 |
2 |
Breaks (isochromatide) |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
Acentric fragments |
3 |
1 |
1 |
0 |
1 |
0 |
2 |
3 |
0 |
1 |
1 |
3 |
Multiple aberrations |
0 |
1 |
0 |
1 |
0 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
Table 2: Testing povidone-iodine in the bone marrow test on Chinese Hamster with five intraperitoneal injection
Dose |
Untreated |
5 x 10 mL of saline/kg |
5 x 38.3 mg/kg |
No of animals |
12 |
11 |
11 |
Analyzed metaphases |
1200 |
1100 |
1100 |
Animals with aberrant metaphases |
7 |
8 |
7 |
Metaphases with aberrations |
|
|
|
Including gaps |
10 |
25 |
13 |
Excluding gaps |
2 |
7 |
2 |
Gaps (chromatide) |
8 |
17 |
8 |
Gaps (isochromatide) |
0 |
1 |
3 |
Breaks (chromatide) |
1 |
6 |
2 |
Breaks (isochromatide) |
0 |
0 |
0 |
Acentric fragments |
0 |
1 |
0 |
Multiple aberrations |
1 |
0 |
0 |
The animals tolerated a single application of povidone-iodine without symptoms. The conception rate decreased significantly during the last week, but the average number of implantations and the mutagenicity index were not affect During the other mating periods, all parameters varied in a similar manner as in the control animals (Table 1).
Table 1: Testing of povidone-iodine in the dominant lethal test on male NMRI mice when applied a single intraperitoneal dose.
|
|
Females with implantations |
Total implantations |
Live embryos |
Dead implantations |
Mutagenicity index |
||||
Dose |
Mating week |
No |
% |
No |
Per female |
No |
Per female |
No |
Per female |
|
0 |
1 |
48 |
80.0 |
548 |
11.4 |
491 |
10.2 |
57 |
1.2 |
10.4 |
2 |
58 |
96.7 |
771 |
13.3 |
705 |
12.2 |
66 |
1.1 |
8.6 |
|
3 |
58 |
96.7 |
750 |
12.9 |
690 |
11.9 |
60 |
1.0 |
8.0 |
|
4 |
56 |
93.3 |
734 |
13.1 |
671 |
12.0 |
63 |
1.1 |
8.6 |
|
5 |
57 |
95.0 |
737 |
12.9 |
666 |
11.7 |
71 |
1.2 |
9.6 |
|
6 |
58 |
96.7 |
736 |
12.7 |
655 |
11.3 |
81 |
1.4 |
11.0 |
|
7 |
58 |
96.7 |
761 |
13.1 |
705 |
12.2 |
56 |
1.0 |
7.4 |
|
8 |
54 |
90.0 |
731 |
13.5 |
652 |
12.1 |
79 |
1.5 |
10.8 |
|
10 mg/kg distilled water |
1 |
43 |
71.7 |
516 |
12.0 |
473 |
11.0 |
43 |
1.0 |
8.3 |
2 |
54 |
90.0 |
721 |
13.4 |
652 |
12.1 |
69 |
1.3 |
9.6 |
|
3 |
57 |
95.0 |
717 |
12.6 |
646 |
11.3 |
71 |
1.2 |
9.9 |
|
4 |
56 |
93.3 |
761 |
13.6 |
671 |
12.0 |
90 |
1.6 |
11.8a |
|
5 |
59 |
98.3 |
776 |
13.2 |
699 |
11.8 |
77 |
1.3 |
9.9 |
|
6 |
58 |
96.7 |
802 |
13.8 |
716 |
12.3 |
86 |
1.5 |
10.7 |
|
7 |
60 |
100.0 |
772 |
12.9 |
716 |
11.9 |
56 |
0.9 |
7.3 |
|
8 |
56 |
93.3 |
779 |
13.9 |
712 |
12.7 |
67 |
1.2 |
8.6 |
|
72 mg/kg povidone-iodine |
1 |
33 |
55.0b |
345 |
10.5 |
296 |
9.0 |
49 |
1.5 |
14.2 |
2 |
58 |
96.8 |
772 |
13.3 |
695 |
12.0 |
77 |
1.3 |
10.0 |
|
3 |
59 |
98.3 |
735 |
12.5 |
653 |
11.1 |
82 |
1.4 |
11.2 |
|
4 |
58 |
96.7 |
772 |
13.3 |
716 |
12.3 |
56 |
1.0 |
7.3 |
|
5 |
59 |
98.3 |
784 |
13.3 |
719 |
12.2 |
65 |
1.1 |
8.3 |
|
6 |
58 |
96.7 |
811 |
14.0 |
744 |
12.8 |
67 |
1.2 |
8.3 |
|
7 |
56 |
93.3 |
679 |
12.1 |
607 |
10.8 |
72 |
1.3 |
10.6a |
|
8 |
57 |
95.0 |
749 |
13.1 |
674 |
11.8 |
75 |
1.3 |
10.0 |
(a) p< 0.05; (b) p< 0.01
The animals tolerated administration of povidone-iodine without symptoms. There was merely a slight increase in polychromatic erythrocytes containing micronuclei, but even the increase was within the normal range.
Table 1: Testing of povidone-iodine in the micronucleus
Dose |
0 |
2 x 10 mL distilled water/kg |
2 x 36 mg PVP-I/kg |
Number of animals |
10 |
10 |
10 |
Analyzed normochromatic erythrocytes |
40,000 |
40,000 |
40,000 |
With MN (%) |
2.15 |
3.17 |
2.52 |
Minimum (%) |
0.12 |
0.12 |
0.15 |
Maximum (%) |
0.35 |
0.45 |
0.35 |
1 MN (%) |
1.90 |
2.82 |
2.35 |
2 MN (%) |
0.22 |
0.30 |
0.15 |
3 MN (%) |
0.00 |
0.05 |
0.00 |
More than 3 MN (%) |
0.02 |
0.00 |
0.02 |
Analyzed polychromatic erythrocytes |
20,000 |
20,000 |
20,000 |
With MN (%) |
3.70 |
2.95 |
4.30 (a) |
Minimum (%) |
0.20 |
0.20 |
0.20 |
Maximum (%) |
0.50 |
0.50 |
0.65 |
1 MN (%) |
3.60 |
2.75 |
4.20 |
2 MN (%) |
0.10 |
0.20 |
0.05 |
3 MN (%) |
0.00 |
0.00 |
0.05 |
More than 3 MN (%) |
0.00 |
0.00 |
0.00 |
MN: Micronucleus(ei); PVP-I: Povidone-iodine.
(a)
p< 0.05
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Iodine is a well-known micronutrient that is essential for the synthesis of thyroid hormones in all vertebrates, as well as a promoter of metamorphosis or transformation of life stages in several invertebrates. Iodine and iodide compounds have been reported to be non-genotoxic in expert reviews (EVM, 2002; WHO, 2009, WHO 2020). In the registration dossier of iodine (including PVP-iodine) as an active substance for biocidal products (Ref: Assessment report, Iodine (including PVP-iodine) Product types 1, 3, 4, 22, 13 December 2013, Sweden), it was concluded that no genotoxic potential of iodine could be identified based on the available in vitro and in vivo data package and an overall weight of evidence assessment.
It has been reported (Hincal 2009) that severe iodine deficiency may impair thyroid hormone synthesis and cause a compensatory increase in hydrogen peroxide concentration in thyrocytes. This increase may result in a more prolonged exposure to oxygen-free radicals, which in turn may lead to DNA damage and induce mutations.
Iodine and chemically related compounds have been tested for genotoxicity in a variety of systems both in vitro and in vivo (Table 1). For this endpoint, data from iodine, iodine tincture and polyvinylpyrrolidone-iodine complex (PVP-I) has been assessed. PVP-I is a complex of polyvinylpyrrolidone (povidone) and iodine, designed to be a stable form of complexed iodine. 9-12 % available iodine is present in standard United States Pharmacopoeia (USP) preparations. PVP-I is used in surface disinfection but is also used extensively in antiseptic preparations of iodine as well as in drug and other applications leading to direct human contact/intake. No consistent or relevant toxic effects in humans have been reported with PVP-I. Iodine tincture is a mixture containing 2 % iodine and 2.0 % sodium iodide in 50% alcohol and is also available as strong iodine tincture (7 % iodine and 5 % KI in 83 % ethanol). There are no reports on the genotoxicity of potassium iodide and on the classification of ethanol under the criteria applied for classification and labelling of chemicals. Both PVP-I and iodine tincture act by releasing iodine to the surrounding medium, thus the potential genotoxic activity may result from the available iodine.
Table 1: Summary of available genotoxicity studies for iodine and chemically related compounds
Refererence (Reliability) |
Assay/Condition |
Result/Test compound |
Wlodowski et al., 1975. (3) |
Gene mutation bacteria/In vitro |
Equivocal/PVP-I |
Hori et al., 2007. (2) |
Chromosome aberrations/In vitro |
Positive/Iodine tincture |
Hori et al., 2007. (2) |
Chromosome aberrations/In vitro |
Negative/PVP-I |
Hikiba et al., 2005. (2) |
Chromosome aberrations/In vitro |
Positive/Iodine |
Miyachi and Tsutsui, 2005. (2) |
Sister chromatid exchanges/In vitro |
Positive/Iodine |
Yamamoto and Tsutsui, 2005. (2) |
Unscheduled DNA synthesis/In vitro |
Negative/Iodine |
Kessler et al., 1980. (2) |
Gene mutation in mamm. cells/In vitro |
Negative/PVP-I |
Kessler et al., 1980. (2) |
Gene mutation in mamm. cells/In vitro |
Negative/Iodine |
Merkle and Zeller, 1979. (2) |
Dominant Lethal Assay/In vivo |
Negative/PVP-I |
Merkle and Zeller, 1979. (2) |
Bone marrow micronucleus test/In vivo |
Negative/PVP-I |
Merkle and Zeller, 1979. (2) |
Bone marrow chromosome aberrations test/In vivo |
Negative/PVP-I |
Hincal (2009), reported that severe iodine deficiency may impair thyroid hormone synthesis and cause a compensatory increase in hydrogen peroxide concentration in thyrocytes. This increase may result in a more prolonged exposure to oxygen-free radicals, which in turn may lead to DNA damage and induce mutations.
Wlodowski et al. (1975) reported that povidone-iodine (PVP-I) was mutagenic in the TA1530 but not in the TA1538 tester strains of Salmonella typhimurium. The positive findings, however, were encountered under unusual temperature conditions (at 4 °C, and not the usual 37 °C in current standards). Also, the applicability of a bacterial system to evaluate the mutagenicity of a bactericide agent remains questionable. Povidone-iodine and iodine were negative in the L5178Y mouse lymphoma assay in the absence of metabolic activation and in the presence of metabolic activation, povidone-iodine showed only marginal activity.
Cytogenicity assays in vitro are consistent with a potential positive effect of iodine, however, it is suggested that iodine may induce chromosome damage by secondary mechanisms associated to cytotoxicity, but not as a result of DNA damage. In vitro mammalian tests have raised concern for the high rate of false positive results obtained with cytotoxic chemicals (Kirkland et al., 2007; Pfuhler et al., 2010). Iodine is in fact a high oxidizing agent which can react with proteins leading to high cytotoxicity in vitro. This hypothesis could be proven by the lack of cytogenotoxicity of PVP-I.
Doses of 72 mg/kg be povidone iodine (given by i.p. injection) were not mutagenic in the mouse dominant lethal assay (Merkle and Zeller, 1979). Povidone-iodine was also tested in mouse micronucleus assay and was negative at doses of 35 mg/kg bw (i.p) and did not produce chromosome aberrations in the in the bone marrow of Chinese hamsters given i.p. doses of 38.3 and 82.5 mg/kg bw, one quarter and one half of the LD50 respectively.
Although positive results are available for iodine in in vitro cytogenetic assays, in vivo results in both somatic (micronucleus and chromosome aberrations tests) and germ cells (dominant lethal test) indicate that iodine can be regarded as a non genotoxic substance. Based on the overall package provided and the fact that in vivo studies are more biologically relevant to humans, no additional studies are considered necessary.
References (not summarized in this section of the IUCLID File):
- Expert Group on Vitamins and Minerals (EVM). Revised review of iodine. 2002.
- Hincal, F. Oxidative damage in iodine deficiency. In: “Comprehensive handbook of iodine: nutritional, biochemical, pathological and therapeutic aspects”. USA, 2009.
- Kirkland, D., Pfuhler, S., Tweats, D., Aardema, M., Corvi, R., Darroudi, F., Elhajouji, A., Glatt, H., Hastwell, P., Hayashi, M., Kasper, P., Kirchner, S., Lynch, A., Marzin, D., Maurici, D., Meunier, J-R., Muller, L., Nohynek, G., Parry, J., Parry, E., Thybaud, V., Tice, R., van Benthem, J., Vanparys, P., White, P. 2007. How to reduce false positive results when undertaking in vitro genotoxicity testing and thus avoid unnecesary follow-up animal tests: Report of an ECVAM Workshop. Mutat. Res. 628: 31 -55.
- Pfuhler, S., Kirst, A., Aardema, M., Banduhn, N., Goebel, C., Araki, D., Costabel-Farkas, Dufour, E., Fautz, R., Harvey, J., Hewitt, N., Hibatallah, J., Carmichael, P., Macfarlane, M., Reisinger, K., Rowland, J., Schellauf, F., Schepky, A., Scheel, J. 2010. A tiered approach to the use of alternatives to animal testing for the safety assessment of cosmetics: Genotoxicity. A COLIPA analysis. Regul. Toxicol. Pharmacol. 57: 315 -324.
- World Health Organization (WHO). Iodine and inorganic iodides: human health aspects. Concise international chemical assessment document: 72. 2009.
- World Health Organization (WHO). Iodine in dinking water. Background document fo development of WHO guidelines for drinking water quality. 2020.
- Registration dossier of iodine (including PVP-iodine) as an active substance for biocidal products; Assessment report, Iodine (including PVP-iodine) Product types 1, 3, 4, 22, 13 December 2013, Sweden)
Short description of key information:
In vitro gene mutation in bacteria: Equivocal
In vitro cytogenicity studies in mammalian cells: Positive
In vitro gene mutation in mammalian cells: Negative
In vivo test, germ cells: Negative
In vivo tests, somatic cells: Negative
Endpoint Conclusion: No adverse effect observed (negative)
Justification for classification or non-classification
Iodine is not classified as a germ cell mutagenic substance according to Regulation (EC) No 1272/2008, based on the lack of genotoxic activity of chemically related compound (polyvinylpirrolidone iodine complex) in both germinal and somatic cells in vivo.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.