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EC number: 204-846-3 | CAS number: 127-51-5
- 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
Genetic toxicity: in vivo
Administrative data
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- January - March 2000
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 000
- Report date:
- 2000
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Version / remarks:
- July, 1997
- Qualifier:
- according to guideline
- Guideline:
- other: International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (1996 and 1997)
- GLP compliance:
- yes
- Type of assay:
- mammalian bone marrow chromosome aberration test
Test material
- Reference substance name:
- Ionone, methyl-
- EC Number:
- 215-635-0
- EC Name:
- Ionone, methyl-
- Cas Number:
- 1335-46-2
- Molecular formula:
- C14H22O
- IUPAC Name:
- (1E)-1-(2,6,6-trimethylcyclohex-2-en-1-yl)pent-1-en-3-one
Constituent 1
Test animals
- Species:
- mouse
- Strain:
- ICR
- Details on species / strain selection:
- ICR mice for the study were obtained from Harlan Sprague Dawley, Inc., Frederick, MD.
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Harlan Sprague Dawley, Inc., Frederick, MD
- Age at study initiation: 6 to 8 weeks old
- Weight at study initiation: Male: 25.0 - 29.6 g; Female: 25.2 - 29.7 g
- Housing: AAALAC-accredited facility: five mice per cage in polycarbonate cages which were maintained on stainless steel racks equipped with automatic watering manifolds and which were covered with filter material. Heat-treated hardwood chips were used for bedding.
- Diet: certified laboratory rodent chow ad libitum
- Water: tab water ad libitum
- Acclimation period: 5 days
ENVIRONMENTAL CONDITIONS
- Temperature: 72 ± 3°F
- Humidity: 50 ± 20% relative humidity
- Photoperiod: 12 hour light/dark cycle
Administration / exposure
- Route of administration:
- intraperitoneal
- Vehicle:
- - Solvent used: corn oil
- Justification for choice of vehicle: compatibility of the vehicle with the test system animals
- Concentration of test material in vehicle: 20 mg/mL - Details on exposure:
- The test article-vehicle mixture, the vehicle alone, or the positive control was administered by intraperitoneal injection at a constant volume of 20mL/kg body weight. Intraperitoneal injection was selected to maximize delivery of the test article to the test system. All mice in the experimental and control groups were weighed immediately prior to dose administration, and the dose volume was based on individual body weights. Mice were observed after dose administration for clinical signs of chemical effect.
- Frequency of treatment:
- Once
- Post exposure period:
- 24 h (462.5 and 925 mg/kg bw); 24 and 48h (1850 mg/kg bw)
Doses / concentrationsopen allclose all
- Dose / conc.:
- 462.5 mg/kg bw/day (nominal)
- Dose / conc.:
- 925 mg/kg bw/day (nominal)
- Dose / conc.:
- 1 850 mg/kg bw/day (nominal)
- No. of animals per sex per dose:
- 5 (462.5 and 925 mg/kg bw); 15 (1850 mg/kg bw)
- Control animals:
- yes
- Positive control(s):
- Positive Control Cyclophosphamide (CP), 50 mg/kg, 5 mice per sex, post exposure period 24 h
Examinations
- Tissues and cell types examined:
- Using oil immersion, 2000 polychromatic erythrocytes were scored for the presence of micronuclei which are defined as round, darkly staining nuclear fragments, having a sharp contour with diameters usually from 1/20 to 1/5 of the erythrocyte. The number of micronucleated normochromatic erythrocytes in the field of 2000 polychromatic erythrocytes was enumerated. The proportion of polychromatic erythrocytes to total erythrocytes was also recorded per 1000 erythrocytes.
- Details of tissue and slide preparation:
- Immediately following sacrifice, the femurs were exposed, cut just above the knee, and the bone marrow was aspirated into a syringe containing fetal bovine serum. The bone marrow cells were transferred to a capped centrifuge tube containing approximately 1 mL fetal bovine serum. The bone marrow cells were pelleted by centrifugation at approximately 100 x g for five minutes and the supernatant was drawn off, leaving a small amount of serum with the remaining cell pellet. The cells were resuspended by aspiration with a capillary pipet and a small drop of bone marrow suspension was spread onto a clean glass slide. Two slides were prepared from each mouse. The slides were fixed in methanol, stained with May-Gruenwald-Giemsa and permanently mounted.
- Evaluation criteria:
- In order to quantify the proliferation state of the bone marrow as an indicator of bone marrow toxicity, the proportion of polychromatic erythrocytes to total erythrocytes was determined for each animal and treatment group.
- Statistics:
- The incidence of micronucleated polychromatic erythrocytes per 2000 polychromatic erythrocytes was determined for each mouse and treatment group. Statistical significance was determined using the Kastenbaum-Bowman tables which are based on the binomial distribution (Kastenbaum and Bowman, 1970). All analyses were performed separately for each sex and sampling time.
Results and discussion
Test results
- Key result
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- The incidence of micronucleated polychromatic erythrocytes per 2000 polychromatic erythrocytes scored and the proportion of polychromatic erythrocytes per total erythrocytes are summarized and presented for each treatment group by sacrifice time in Table 1. Individual animal data are presented in Tables 2 and 3. Slight to moderate reductions of 6% to 35% in the ratio of polychromatic erythrocytes to total erythrocytes were observed in some of the test article-treated groups relative to their respective vehicle controls. Reductions were observed in male and female dose groups 24 hours after treatment with 462.5, 925 and 1850 mg/kg and in female mice 48 hours after treatment with 1850 mg/kg. The reduction in the frequency of polychromatic erythrocytes in the bone marrow suggest that there was bioavailability of the test article to the bone marrow target tissue. A statistically significant increase in micronucleated polychromatic erythrocytes (8 MNPCEV10,000 PCE) in the test article treated group relative to the respective vehicle control group was observed in male mice at 24 hours after treatment with 925 mg/kg (p<0.05, Kastenbaum-Bowman Tables). However, this response is not considered biologically relevant since each of the five animals had no more then 3 MNPCE that are within the range of the historical solvent control (0-7 MN/2000 PCE/animal). No significant increase and no dose-responsive increase was observed in any other test article treated group regardless of dose level, sex, or bone marrow collection time. CP induced a significant increase in micronucleated polychromatic erythrocytes in both male and female mice (p<0.05, Kastenbaum-Bowman Tables).
Any other information on results incl. tables
Table 1:Summary of Bone Marrow Micronucleus Study Using Methyl ionone
Treatment |
Sex |
Time (hr) |
Number of Mice |
PCE/Total Erythrocytes (Mean ± SD) |
Change from Control (%) |
Micronucleated Polychromatic Number per 1000 PCEs (Mean ± SD) |
Erythrocytes Number per PCEs Scored1 |
Corn oil 20 mL/kg
|
M F |
24 24 |
5 5 |
0.463 ± 0.04 0.489 ± 0.07 |
--- --- |
0.1 ± 0.22 0.4 ± 0.42 |
1 / 10000 4 / 10000 |
Methyl ionone 462.5 mg/kg
|
M F |
24 24 |
5 5 |
0.363 ± 0.06 0.377 ± 0.04 |
-22 -23 |
0.2 ± 0.27 0.2 ± 0.27 |
2 / 10000 2 / 10000 |
925 mg/kg |
M F |
24 24 |
5 5 |
0.344 ± 0.06 0.339 ± 0.04 |
-26 -31 |
0.8 ± 0.45 0.5 ± 0.50 |
*8 / 10000 5 / 10000 |
1850 mg/kg |
M F |
24 24 |
5 5 |
0.434 ± 0.07 0.319 ± 0.09 |
-6 -35 |
0.3 ± 0.27 0.7 ± 0.45 |
3 / 10000 7 / 10000 |
CP 50 mg/kg
|
M F |
24 24 |
5 5 |
0.334 ± 0.01 0.329 ± 0.07 |
-28 -33 |
27.8 ± 4.19 40.4 ± 9.90 |
*278 / 10000 *404 / 10000 |
Corn oil 20 mL/kg
|
M F |
48 48 |
5 5 |
0.451 ± 0.07 0.485 ± 0.03 |
--- --- |
0.1 ± 0.22 0.2 ± 0.27 |
1 / 10000 2 / 10000 |
Methyl ionone 1850 mg/kg
|
M F |
48 48 |
5 5 |
0.455 ± 0.03 0.385 ± 0.07 |
1 -21 |
0.1 ± 0.22 0.2 ± 0.27 |
1 / 10000 2 / 10000 |
1* p≤0.05 (Kastenbaum-Bowman Tables)
Table 2: Induction of Micronucleated Polychromatic Erythroxcytes in Bone Marrow Cells collected 24 hours after a single dose of Methyl ionone
Treatment |
Sex |
Animal Number |
PCE / Total Erythrocytes |
Micronucleated PCE (Number / PCE scored) |
Corn oil
20 ml/kg |
M |
101 102 103 104 105 |
0.413 0.531 0.435 0.475 0.461 |
0 / 2000 0 / 2000 0 / 2000 0 / 2000 1 / 2000 |
F |
106 107 108 109 110 |
0.486 0.379 0.510 0.556 0.512 |
0 / 2000 1 / 2000 2 / 2000 0 / 2000 1 / 2000 |
|
Methyl ionone
462.5 mg/kg |
M |
111 112 113 114 115 |
0.334 0.442 0.360 0.400 0.277 |
1 / 2000 0 / 2000 0 / 2000 1 / 2000 0 / 2000 |
F |
116 117 118 119 120 |
0.382 0.387 0.310 0.418 0.387 |
1 / 2000 1 / 2000 0 / 2000 0 / 2000 0 / 2000 |
|
925 mg/kg |
M |
121 122 123 124 125 |
0.259 0.330 0.411 0.406 0.312 |
1 / 2000 3 / 2000 2 / 2000 1 / 2000 1 / 2000 |
F |
126 127 128 129 130 |
0.336 0.389 0.306 0.372 0.293 |
1 / 2000 2 / 2000 0 / 2000 2 / 2000 0 / 2000 |
|
1850 mg/kg |
M |
131 132 133 134 135 |
0.500 0.494 0.426 0.329 0.421 |
0 / 2000 1 / 2000 0 / 2000 1 / 2000 1 / 2000 |
F |
136 137 138 139 140 |
0.205 0.325 0.327 0.289 0.449 |
2 / 2000 0 / 2000 2 / 2000 1 / 2000 2 / 2000 |
|
CP
50 mg/kg |
M |
141 142 143 144 145 |
0.346 0.342 0.318 0.345 0.320 |
66 / 2000 59 / 2000 44 / 2000 51 / 2000 58 / 2000 |
F |
146 147 148 149 150 |
0.334 0.384 0.219 0.374 0.332 |
73 / 2000 50 / 2000 99 / 2000 88 / 2000 94 / 2000 |
Table 3: Induction of Micronucleated Polychromatic Erythrocytes in Bone Marrow Cells collected 48 hours after a single dose of Methyl ionone
Treatment |
Sex |
Animal Number |
PCE / Total Erythrocytes |
Micronucleated PCE (Number / PCE scored) |
Corn oil
20 mL/kg |
M |
151 152 153 154 155 |
0.377 0.555 0.468 0.435 0.419 |
0 / 2000 1 / 2000 0 / 2000 0 / 2000 0 / 2000 |
F |
156 157 158 159 160 |
0.499 0.459 0.472 0.459 0.536 |
1 / 2000 0 / 2000 0 / 2000 1 / 2000 0 / 2000 |
|
Methyl ionone
1850 mg/kg |
M |
*171 162 163 164 165 |
0.492 0.426 0.478 0.443 0.438 |
0 / 2000 0 / 2000 1 / 2000 0 / 2000 0 / 2000 |
F |
166 167 168 169 170 |
0.400 0.327 0.461 0.431 0.307 |
0 / 2000 1 / 2000 0 / 2000 0 / 2000 1 / 2000 |
* = Replacement animal
Applicant's summary and conclusion
- Conclusions:
- The test item was negative in the mouse micronucleus assay (OECD 474).
- Executive summary:
The test item was tested in the mouse micronucleus assay according to OECD 474. The assay was performed in two phases. The first phase, designed to set dose levels for the definitive study, consisted of a pilot toxicity study followed by a toxicity study. The second phase, the micronucleus study, evaluated the potential of the test article to increase the incidence of micronucleated polychromatic erythrocytes in bone marrow of male and female mice. In both phases of the study, test and control articles were administered in a constant volume of 20 mL/kg body weight by a single intraperitoneal injection. Corn oil was determined to be the solvent of choice based on the compatibility of the vehicle with the test system animals. The test article was soluble in corn oil at 20 mg/mL, the maximum concentration tested in the study. Dosing concentrations were delivered to the test system as clear light yellow solutions. In the pilot toxicity study, male mice were dosed with 1, 10, 100, or 1000 mg test article/kg body weight and male and female mice were dosed with 2000 mg/kg. Mortality was observed in 1/5 male mice and 2/5 female mice at 2000 mg/kg. Clinical signs following dose administration included lethargy and piloerection in males at 1000 mg/kg and in males and females at 2000 mg/kg. Convulsions were observed in males and females at 2000 mg/kg and prostration and crusty eyes were observed in female mice at 2000 mg/kg. In the toxicity assay, male and female mice were dosed with 1500 and 1750 mg test article/kg body weight. No mortality was observed in any male or female mice during the course of the study. Clinical signs following dose administration included lethargy and piloerection in males and females at 1500 and 1750 mg/kg. Due to the mortality at 2000 mg/kg in the pilot toxicity study, the high dose for the micronucleus test was set at 1850 mg/kg, which was estimated to be the maximum tolerated dose. In the micronucleus assay, male and female mice were dosed with 462.5, 925 or 1850 mg/kg body weight. Mortality was observed only in 1/15 male mice receiving 1850 mg/kg. This animal was replaced at the time of bone marrow collection with a replacement animal that also received 1850 mg/kg. All treatment groups had five animals for bone marrow analysis. Clinical signs following dose administration included: lethargy and piloerection in males and females at 925 and 1850 mg/kg and crusty eyes in male and female mice at 1850 mg/kg. Bone marrow cells, collected 24 and 48 hours after treatment, were examined microscopically for micronucleated polychromatic erythrocytes. Slight to moderate reduction (up to 35%) in the ratio of polychromatic erythrocytes to total erythrocytes were observed in some of the test article-treated groups relative to the respective vehicle controls. These reductions suggest bioavailability of the test article to the bone marrow target. A statistically significant increase in micronucleated polychromatic erythrocytes (8 MNPCE/10000 PCE) in test article-treated group relative to the respective vehicle control group was observed in male mice 24 after treatment with 925 mg/kg (p<0.05, Kastenbaum-Bowman). However, this response is not considered biologically relevant, since each of the five animals had no more than 3 MPCE. These numbers of MNPCE are within the range of historical solvent control (0-7 MN/2000 PCE/animal). No significant increase and no dose responsive increase was observed in any other test article treated group regardless of dose level, sex, or bone marrow collection time. The results of the assay indicate that under the conditions described in this report, the test item did not induce a significant increase in micronucleated polychromatic erythrocytes in either male or female mice. The test item was concluded to be negative in the mouse micronucleus assay.
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