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EC number: 460-110-3 | CAS number: 797751-43-0 WASOX-MMAC2
- 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
In vitro bacterial gene mutation: Key study: OECD Guideline 471 and GLP. The test substance did not show any mutagenic activity.
In vitro cytogenicity in mammalian cells: Key study: OECD Guideline 473 and GLP. The test substance did not show any clastogenic activity.
In vitro gene mutation in mammalian cells: Key study: OECD Guideline 476 and GLP. Based on read-across approach from the analogue acetone oxime, Wasox-MMAC2 was determined not to be mutagenic.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- October 11 - November 5, 2004
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- According to OECD Guideline 471, with GLP.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- histidine-requiring gene
- Species / strain / cell type:
- S. typhimurium TA 1535
- Species / strain / cell type:
- S. typhimurium TA 98
- Species / strain / cell type:
- S. typhimurium TA 100
- Species / strain / cell type:
- S. typhimurium TA 102
- Species / strain / cell type:
- other: S. typhimurium TA 97a
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 liver microsomal fraction
- Test concentrations with justification for top dose:
- 5000, 1667, 556, 185, and 62 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: the test substance hydrolyses rapidly in water - Negative solvent / vehicle controls:
- yes
- Remarks:
- (DMSO)
- Positive controls:
- yes
- Remarks:
- (without activation)
- Positive control substance:
- other: 4-nitro-o-phenylene-diamine
- Remarks:
- TA97a: 4-nitro-o-phenylene-diamine- 10 µg
- Positive controls:
- yes
- Remarks:
- (with activation)
- Positive control substance:
- 7,12-dimethylbenzanthracene
- Remarks:
- TA97a:7,12-dimethylbenz(a)anthracene-10 µg
- Positive controls:
- yes
- Remarks:
- (without activation)
- Positive control substance:
- 2-nitrofluorene
- Remarks:
- TA98: 2-nitrofluorene - 2 µg
- Positive controls:
- yes
- Remarks:
- (without activation)
- Positive control substance:
- sodium azide
- Remarks:
- TA100: sodium azide-2 µg and TA1535: sodium azide- 1 µg
- Positive controls:
- yes
- Remarks:
- (with activation)
- Positive control substance:
- other: 2- aminoanthracene
- Remarks:
- TA98:2- aminoanthracene- 1 µg , TA1535: aminoanthracene- 2 µg and TA100: aminoanthracene- 2 µg
- Positive controls:
- yes
- Remarks:
- (with activation)
- Positive control substance:
- other: 1,8-Dihydroxy-anthraquinone
- Remarks:
- TA102: 1,8-Dihydroxy-anthraquinone - 50 µg
- Positive controls:
- yes
- Remarks:
- (without activation)
- Positive control substance:
- other: t-Butyl-hydroperoxide
- Remarks:
- TA102: t-Butyl-hydroperoxide-50 µg
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation). Bacteria, test substance (and microsomes) are in contact on the plate without preceding incubation in the liquid state. For each sample the following solutions were combined:
- 0.1 mL of the overnight culture of the bacteria,
- 0.5 mL of S9-mix (or phosphate buffered saline for samples without metabolic activation),
- 0.1 mL of the appropriate test- or reference substance solution and
- 2 mL of top agar.
DURATION
- Exposure duration: 48 h
NUMBER OF REPLICATIONS: triplicates (an independent repetition of the experiment was performed)
DETERMINATION OF CYTOTOXICITY
Method: the citotoxicity was rated as follows:
A reduced bacterial background lawn (mottled instead of homogeneous).
Microcolonies of bacteria instead of a homogeneous background lawn.
No background lawn.
Clearly reduced numbers of revertant colonies.
- Evaluation criteria:
- The criteria for a positive result are:
A reproducible increase of the number of revertants to more than the following threshold values for at least one of the concentrations:
- For the strains with a low spontaneous revertant rate i.e. TA98 and TA1535: The 2.5 fold of the amount of the spontaneous revertants.
- For the strains with a high spontaneous revertant rate i.e. TA97a, TA100 and TA102: The 1.67 fold of the amount of the spontaneous revertants.
These threshold values were derived from the variations in the control samples of the Ames test. - Key result
- Species / strain:
- other: TA97a, TA98, TA100, TA102, and TA1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No precipitation of the test substance was seen in any on the concentrations groups
RANGE-FINDING/SCREENING STUDIES:
Preliminary experiment was performed: different concentrations of test substance solutions were mixed with phosphate buffer, bacteria (TA100) and top-agar, as described in 3.5 and spread over a plate with minimal agar. The plates were incubated at 37 °C for 2 days and the growth of the bacterial background and the density of revertant colonies was determined.
COMPARISON WITH HISTORICAL CONTROL DATA:
The numbers of spontaneous revertants were comparable with the historic control data for the negative controls.
- Conclusions:
- According to the results obtained in this study, the test substance is non-mutagenic in the Ames test with the Salmonella typhimurium strains TA97a, TA98, TA100, TA102 and TA1535 with and without an external metabolising system up to 5000 µg/plate.
- Executive summary:
The Bacterial Reverse mutation Assay (Ames test) for the test substance Wasox-MMAC2 was performed according to OECD Guideline 471 with five histidine dependents strains of Salmonella typhimurium TA97a, TA98, TA100, TA102 and TA1535. Plate Incorporation Method was carried out at the concentrations of 5000, 1667, 556, 185, and 62 µg/plate in the presence and absence of metabolic activation. No precipitation of the test substance and no signs of cytotoxicity were noted at any dose level. In none of the concentrations tested and with none of the strains used an significant increase of the mutation frequency to more than the threshold values (250 % of the controls for strains TA98 and TA1535 and 167 % of the controls for strains TA97a, TA100 and TA102) was obtained. Metabolic activation did not change these results. According to the obtained results, the test substance is non-mutagenic in the Ames test with and without an external metabolizing system up to 5000 µg/plate.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- November 24, 2004 - March 24, 2005
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- According to OECD guideline 473, with GLP.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- lymphocytes:
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- 5.000, 1.670, 0.560, and 0.185 µL/mL
- Untreated negative controls:
- yes
- Remarks:
- The culture medium
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- Without metabolic activation system
- Untreated negative controls:
- yes
- Remarks:
- The culture medium
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- With metabolic activation system
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium. 10 mL of RPMI medium (3 hours treatment) or of complete culture medium (for 20 hours treatment) containing the test substance at the appropriate concentrations. Each culture with the use of a metabolic activation system contained 5% S9-Mix (v/v)
- Evaluation criteria:
- A statistically significant increase in the number of metaphases with aberrations or a concentration-relate increase in this number is considered as a positive result. However, a result can also be regarded as positive when other than merely statistical considerations, for example, the kind of aberration are taken into account.
- Statistics:
- The Chi2-Test (two-tailed, p<0.05) was used for the comparison between the negative control and the substance cultures. If the results were positive, comparisons were made separately between the negative control and each concentration. If conditions for the Chi2-Test were not met, Fisher’s Exact Test was used. Chi2-Test or Fisher’s Exact Test were also used for the comparison between the negative and the positive controls.
- Key result
- Species / strain:
- lymphocytes: (3 h treatment)
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- (at 5.00 µL/mL)
- Vehicle controls validity:
- not applicable
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- lymphocytes: (3 h treatment)
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not applicable
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- lymphocytes: (20 h treatment)
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- (at 5.00 and 1.67 µL/mL)
- Vehicle controls validity:
- not applicable
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- COMPARISON WITH HISTORICAL CONTROL DATA:
All figures were within the range of historical negative controls. - Conclusions:
- Under study conditions, there was no relevant evidence that the test substance Wasox-MMAC2 did induce structural chromosomal aberrations in cultured human Iymphocytes, neither without nor with the use of a metabolic activation system.
- Executive summary:
An in vitro Mammalian Chromosome Aberration Test in human lymphocytes was performed according to OECD Guideline 473 to determine possible mutagenic properties of the test substance. A total of four experiments were performed: two of them without and two with the use of a metabolic activation system. A concentration range between 5.000 and 0.185 µL of test substance per mL of medium was tested. The test substance caused marked and severe cytotoxic effects at the concentrations of 5.00 (3 and 20 h of treatment) and 1.67 (20 h of treatment) µL/mL medium when no metabolic activation system was used. There was, under the conditions of this study, no relevant evidence that "WASQX-MMAC2" did induce structural chromosomal aberrations in cultured human lymphocytes, neither without nor with the use of a metabolic activation system. The conclusion is based on a lack of a statistically significant increase of metaphases with structural aberrations in any experiment and on the finding that all figures were within the range of historical negative controls.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- July 10, 2012 - August 23, 2012
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Remarks:
- Test method according to EU Method B.17 and OECD Guideline 476. GLP study.
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian cell gene mutation test using the Hprt and xprt genes
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-mix (Rat Liver Homogenate S9 Fraction)
- Test concentrations with justification for top dose:
- Assay 1, 3-hour treatment with metabolic activation: 5000; 3750; 2500; 1250; 625 and 312.5 μg/mL
Assay 1, 3-hour treatment without metabolic activation: 5000; 3750; 2500; 1250; 625 and 312.5 μg/mL
Assay 2, 3-hour treatment with metabolic activation: 5000; 3750; 2500; 1250; 625 and 312.5 μg/mL
Assay 2, 24-hour treatment without metabolic activation: 5000; 4000; 3000; 2000; 1000; 500; 250 and 125 μg/mL - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Based on the result of the preliminary solubility test the test item was insoluble in Distilled water at 500 mg/mL concentration, but it was soluble in Dimethyl sulfoxide (DMSO) at the same concentrations. - Untreated negative controls:
- yes
- Remarks:
- Untreated control
- Negative solvent / vehicle controls:
- yes
- Remarks:
- (DMSO 1% v/v)
- Positive controls:
- yes
- Remarks:
- without metabolic activation
- Positive control substance:
- 4-nitroquinoline-N-oxide
- Remarks:
- 0.15 μg/mL (3h treatment) and 0.1 μg/mL (24h treatment) in DMSO
- Positive controls:
- yes
- Remarks:
- with metabolic activation
- Positive control substance:
- cyclophosphamide
- Remarks:
- 4 μg/mL in DMSO
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium (RPMI-5 medium)
DURATION
- Exposure duration: Assay 1: 3h (with and without metabolic activation); Assay 2: 3h (with metabolic activation), 24h (without metabolic activation)
- Expression time (cells in growth medium, to allow expresion of the TK- mutation ): 3 days
- Selection time (plating for -trifluorothymidine (TFT) resistance): Two weeks
SELECTION AGENT (mutation assays): 5-trifluorothymidine (TFT)
NUMBER OF REPLICATIONS: Duplicate cultures were used for each treatment
NUMBER OF CELLS EVALUATED:
DETERMINATION OF CYTOTOXICITY
- Method: Cytotoxicity was measured by relative survival
OTHER EXAMINATIONS:
- Determination of Survival or Viability:
After the exposure period and the expression time, cells were also diluted to be plated for survival and viability respectively. Microplates were incubated at 37 ºC ± 0.5 °C containing approximately 5% (v/v) CO2 in air for approximately two weeks. Wells containing viable clones were identified by eye using background illumination and counted.
Parameter calculated:
- Relative survival
- Mutant Frequency
- Small and large colony mutant frequencies were calculated. - Evaluation criteria:
- The test item was considered to be mutagenic if all the following criteria were met:
1. Statistically significant (p < 0.05) and biologically relevant increases in mutation frequency observed in treated cultures compared to the corresponding negative (solvent) control values at one or more concentrations.
3. Increases in mutation frequency reproducible between replicate cultures and/or between tests (under the same treatment conditions).
4. Significant concentration-relationship indicated by the linear trend analysis (p < 0.05).
5. Mutation frequency at the test concentration showing the largest increase at least 126 mutants per 106 viable cells (GEF = the Global Evaluation Factor) higher than the corresponding negative (solvent) control value.
Results, which only partially satisfied the acceptance and evaluation criteria, were evaluated on a case-by-case basis. - Statistics:
- Statistical significance of mutant frequencies was performed using Microsoft Excel software. The control log mutant frequency (LMF) was compared to the LMF from each treatment dose, based on Dunnett's test for multiple comparisons and the data checked for a linear trend in mutant frequency with treatment dose using weighted regression. The test for linear trend was one-tailed, therefore negative trend was not considered significant. These tests required the calculation of the heterogeneity factor to obtain a modified estimate of variance.
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS: No insolubility was detected in the final treatment medium at the beginning and end of the treatment in any of the experiments. There were no large changes in pH or osmolality after treatment.
RANGE-FINDING/SCREENING STUDIES: No insolubility, but cytotoxicity was observed in the preliminary experiment in one case (24-hour treatment without metabolic activation). The recommended maximum concentration of 5000 μg/mL was selected as highest examined concentration in the main experiments in each case. The lower test concentrations were generally separated by factor of two. More closely spaced concentrations were used in the expected cytotoxic concentration range to cover the concentration range from the maximum cytotoxicity (resulting approximately 10-20% relative survival) to little or no cytotoxicity.
COMPARISON WITH HISTORICAL CONTROL DATA: The positive controls (Cyclophosphamide in the presence of metabolic activation and 4-Nitroquinoline-N-oxide in the absence of metabolic activation) gave the anticipated increases in mutation frequency over the controls and were in accordance with historical data in all assays.
ADDITIONAL INFORMATION ON CYTOTOXICITY: No cytotoxicity was observed in the main test up to 5000 µg/mL. - Conclusions:
- No mutagenic effect of Acetonoxim was observed either in the presence or in the absence of metabolic activation system under the conditions of this Mouse Lymphoma Assay.
- Executive summary:
An in vitro mammalian cell assay was performed in mouse lymphoma L5178Y TK+/- 3.7.2 C cells at the tk locus to test the potential of the analogue substance acetone oxime to cause gene mutation and/or chromosome damage. Treatment was performed for 3 hours with and without metabolic activation (±S9 mix) and for 24 hours without metabolic activation (-S9 mix). Dimethyl sulfoxide was used as the solvent of the test item in this study. The test item was examined up to 5000 μg/mL in the Preliminary Toxicity Test. Based on the results of the preliminary experiment, the following test item concentrations were examined in the mutation assays: Assay 1, 3-hour treatment with and without metabolic activation: 5000; 3750; 2500; 1250; 625 and 312.5 μg/mL; Assay 2, 3-hour treatment with metabolic activation: 5000; 3750; 2500; 1250; 625 and 312.5 μg/mL and 24-hour treatment without metabolic activation: 5000; 4000; 3000; 2000; 1000; 500; 250 and 125 μg/mL In Assays 1 and 2, no insolubility was detected in the final treatment medium at the beginning and end of the treatment in any of the experiments. There were no large changes in pH or osmolality after treatment. In both assays no cytotoxicity of the test item was observed. Therefore, an evaluation was made using data of all the six examined concentrations. No biologically relevant or statistically significant increase in the mutation frequency was observed at the evaluated concentrations. No significant dose response to the treatment was indicated by the linear trend analysis. All the validity criteria were fulfilled and the overall study was considered to be valid. In conclusion, no mutagenic effect of Acetonoxim was observed either in the presence or in the absence of metabolic activation system under the conditions of this Mouse Lymphoma Assay.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Wasox-MMAC2 undergoes rapid hydrolysis in aqueous to acetone oxime and the corresponding silanols. These silanols undergo continuous condensation reactions to produce higher molecular weight siloxanes which are considered biologically unavailable. Therefore, the observed toxicity is likely due to the acetone oxime and their values are comparable.
See attached the reporting format and read-across rationale. - Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Based on read-across approach, Wasox-MMAC2 was determined to be not mutagenic under test conditions.
- Conclusions:
- No mutagenic effect of Acetonoxim was observed either in the presence or in the absence of metabolic activation system under the conditions of this Mouse Lymphoma Assay. Based on read-across approach, Wasox-MMAC2 was determined to be not mutagenic under test conditions.
- Executive summary:
An in vitro mammalian cell assay was performed in mouse lymphoma L5178Y TK+/- 3.7.2 C cells at the tk locus to test the potential of the analogue substance acetone oxime to cause gene mutation and/or chromosome damage. Treatment was performed for 3 hours with and without metabolic activation (±S9 mix) and for 24 hours without metabolic activation (-S9 mix). Dimethyl sulfoxide was used as the solvent of the test item in this study. The test item was examined up to 5000 μg/mL in the Preliminary Toxicity Test. Based on the results of the preliminary experiment, the following test item concentrations were examined in the mutation assays: Assay 1, 3-hour treatment with and without metabolic activation: 5000; 3750; 2500; 1250; 625 and 312.5 μg/mL; Assay 2, 3-hour treatment with metabolic activation: 5000; 3750; 2500; 1250; 625 and 312.5 μg/mL and 24-hour treatment without metabolic activation: 5000; 4000; 3000; 2000; 1000; 500; 250 and 125 μg/mL In Assays 1 and 2, no insolubility was detected in the final treatment medium at the beginning and end of the treatment in any of the experiments. There were no large changes in pH or osmolality after treatment. In both assays no cytotoxicity of the test item was observed. Therefore, an evaluation was made using data of all the six examined concentrations. No biologically relevant or statistically significant increase in the mutation frequency was observed at the evaluated concentrations. No significant dose response to the treatment was indicated by the linear trend analysis. All the validity criteria were fulfilled and the overall study was considered to be valid. In conclusion, no mutagenic effect of Acetonoxim was observed either in the presence or in the absence of metabolic activation system under the conditions of this Mouse Lymphoma Assay. Based on these results, the read-across approach was applied and the target substance Wasox-MMAC2 was determined to be not mutagenic under test conditions.
Referenceopen allclose all
Individual numbers of revertants per plates
StrainTA97a: experiment no.1
Concentration |
Revertants |
Revertants |
||||
5000 |
34 |
53 |
40 |
62 |
85 |
81 |
1667 |
64 |
61 |
79 |
128 |
82 |
88 |
556 |
72 |
70 |
59 |
90 |
111 |
123 |
185 |
82 |
70 |
104 |
139 |
96 |
116 |
62 |
61 |
58 |
58 |
125 |
88 |
111 |
solvent |
41 |
94 |
66 |
98 |
82 |
78 |
solvent |
70 |
94 |
70 |
133 |
102 |
110 |
positive |
219 |
1 |
200 |
517 |
482 |
460 |
StrainTA97a: experiment no.2
Concentration |
Revertants |
Revertants |
||||
5000 |
166 |
133 |
136 |
148 |
128 |
145 |
1667 |
111 |
119 |
111 |
149 |
111 |
128 |
556 |
140 |
140 |
136 |
149 |
111 |
131 |
185 |
123 |
137 |
148 |
151 |
154 |
128 |
62 |
123 |
136 |
123 |
165 |
192 |
183 |
solvent |
123 |
142 |
131 |
148 |
151 |
168 |
solvent |
133 |
175 |
123 |
125 |
119 |
145 |
positive |
465 |
675 |
569 |
645 |
526 |
601 |
solvent: DMSO
positive: without metabolisation: 4-Nitro-o-phenylene-diamine, 10 µg / plate
with metabolisation: 7,12-Dimethylbenz[a]anthracene, 10 µg / plate
StrainTA98: experiment no.1
Concentration |
Revertants |
Revertants |
||||
5000 |
9 |
6 |
9 |
13 |
9 |
11 |
1667 |
11 |
11 |
9 |
5 |
9 |
9 |
556 |
8 |
12 |
7 |
4 |
9 |
8 |
185 |
7 |
10 |
10 |
10 |
11 |
13 |
62 |
15 |
10 |
9 |
3 |
11 |
13 |
solvent |
11 |
10 |
10 |
9 |
12 |
12 |
solvent |
6 |
8 |
9 |
14 |
6 |
12 |
positive |
155 |
131 |
10 |
216 |
227 |
209 |
StrainTA98: experiment no.2
Concentration |
Revertants |
Revertants |
||||
5000 |
8 |
6 |
8 |
10 |
4 |
9 |
1667 |
10 |
8 |
12 |
13 |
10 |
19 |
556 |
12 |
9 |
6 |
5 |
15 |
14 |
185 |
8 |
9 |
10 |
9 |
7 |
10 |
62 |
8 |
14 |
15 |
10 |
10 |
15 |
solvent |
13 |
10 |
8 |
10 |
13 |
14 |
solvent |
11 |
5 |
4 |
12 |
11 |
16 |
positive |
343 |
450 |
306 |
338 |
273 |
329 |
solvent: DMSO
positive: without metabolisation: 2-Nitrofluorene, 2 µg / plate
with metabolisation: 2-Amino-anthracene, 1 µg / plate
StrainTA100: experiment no.1
Concentration |
Revertants |
Revertants |
||||
5000 |
44 |
44 |
38 |
78 |
76 |
110 |
1667 |
76 |
67 |
84 |
114 |
111 |
98 |
556 |
76 |
79 |
93 |
87 |
85 |
85 |
185 |
88 |
84 |
105 |
84 |
84 |
84 |
62 |
93 |
90 |
101 |
81 |
93 |
116 |
solvent |
73 |
59 |
88 |
102 |
101 |
122 |
solvent |
66 |
82 |
102 |
113 |
108 |
73 |
positive |
410 |
399 |
375 |
1096 |
1321 |
1081 |
StrainTA100: experiment no.2
Concentration |
Revertants |
Revertants |
||||
5000 |
64 |
47 |
38 |
56 |
62 |
59 |
1667 |
72 |
69 |
81 |
55 |
66 |
76 |
556 |
94 |
94 |
73 |
81 |
69 |
87 |
185 |
66 |
87 |
75 |
105 |
69 |
67 |
62 |
79 |
59 |
76 |
93 |
117 |
82 |
solvent |
87 |
59 |
69 |
84 |
91 |
81 |
solvent |
87 |
84 |
67 |
94 |
84 |
76 |
positive |
181 |
149 |
160 |
469 |
393 |
375 |
solvent: DMSO
positive: without metabolisation: Sodium azide, 2 µg / plate
with metabolisation: 2 -Amino-anthracene, 2 µg / plate
StrainTA102: experiment no.1
Concentration |
Revertants |
Revertants |
||||
5000 |
139 |
128 |
146 |
175 |
261 |
253 |
1667 |
174 |
172 |
136 |
229 |
245 |
203 |
556 |
131 |
137 |
177 |
259 |
236 |
236 |
185 |
155 |
159 |
210 |
230 |
245 |
221 |
62 |
159 |
163 |
134 |
279 |
259 |
282 |
solvent |
169 |
162 |
157 |
251 |
250 |
265 |
solvent |
169 |
206 |
187 |
248 |
239 |
241 |
positive |
629 |
742 |
634 |
629 |
588 |
576 |
StrainTA102: experiment no.2
Concentration |
Revertants |
Revertants |
||||
5000 |
216 |
198 |
213 |
253 |
236 |
244 |
1667 |
172 |
209 |
174 |
251 |
273 |
235 |
556 |
197 |
203 |
197 |
265 |
261 |
235 |
185 |
192 |
219 |
195 |
251 |
241 |
256 |
62 |
198 |
323 |
204 |
241 |
261 |
255 |
solvent |
183 |
206 |
183 |
242 |
267 |
224 |
solvent |
157 |
183 |
148 |
238 |
247 |
248 |
positive |
975 |
902 |
666 |
477 |
442 |
533 |
solvent: DMSO
positive: without metabolisation: t-Butyl-hydroperoxide, 50 µg / plate
with metabolisation: 1,8-Dihydroxy-anthraquinone, 50 µg / plate
StrainTA1535: experiment no.1
Concentration |
Revertants |
Revertants |
||||
5000 |
10 |
11 |
7 |
9 |
12 |
6 |
1667 |
11 |
6 |
15 |
10 |
7 |
10 |
556 |
12 |
7 |
12 |
8 |
13 |
12 |
185 |
10 |
10 |
13 |
10 |
5 |
9 |
62 |
6 |
14 |
8 |
11 |
17 |
10 |
solvent |
6 |
10 |
6 |
12 |
5 |
8 |
solvent |
8 |
4 |
8 |
10 |
15 |
10 |
positive |
256 |
229 |
207 |
181 |
127 |
175 |
StrainTA1535: experiment no.2
Concentration |
Revertants |
Revertants |
||||
5000 |
5 |
14 |
12 |
16 |
18 |
12 |
1667 |
11 |
10 |
15 |
12 |
11 |
14 |
556 |
18 |
12 |
12 |
8 |
13 |
7 |
185 |
16 |
9 |
7 |
6 |
6 |
12 |
62 |
12 |
15 |
11 |
16 |
14 |
11 |
solvent |
14 |
18 |
12 |
18 |
12 |
11 |
solvent |
14 |
18 |
11 |
10 |
12 |
13 |
positive |
253 |
244 |
274 |
154 |
154 |
184 |
solvent: DMSO
positive: without metabolisation: Sodium azide, 1 µg / plate
with metabolisation: 2-Amino-anthracene, 2 µg / plate
Concurrent Positive Controls
The mitotic indices were between 61.1 % and 103.8 % of the negative controls. The positive control substances caused in each experiment clearly higher numbers of metaphases with structural aberrations (statistically significant) than found in the negative controls, without as well as with the use of a metabolic activation system. Additional statistically significant differences to the negative controls like the number of gaps, the number of chromatid-type aberrations.
Mitotic index
In the test substance cultures without a metabolic activation system the mitotic indices were markedly reduced to 54.6 % of the corresponding negative controls after 3 hours of treatment with 5.00 µL/mL. After 20 hours of treatment with 5.00 µL/mL they were reduced to 6.5 % of the negative controls and to 44.9 % at 1.67 µL/mL. At all other concentrations the mitotic indices were between 70.5 and 100.0 % of those of the corresponding negative controls. In the experiments with a metabolic activation system the mitotic indices were in the test substance treated cultures between 89.7 % and 103.6 % of those of the corresponding negative controls.
Structural aberrations
No statistically significant increases in the number of metaphases with structural aberrations were noted in any experiment performed at any concentration analysed compared to the concurrent negative controls, regardless whether a metabolic activation system was used or not. All figures were within or very close to the range of historical negative controls.
Gaps
No statistically significant increases in the number of gaps were noted at any concentration analysed compared to the concurrent negative controls, neither without nor with the use of a metabolic activation system. All figures were within or very close to the range of historical negative controls.
Numerical aberrations
No statistically significant differences in the number of metaphases with numerical aberrations were noted in any experiment performed at any concentration analysed compared to the concurrent negative controls, regardless whether a metabolic activation system was used or not.
In Assay 1 and 2, both with and without metabolic activation, no cytotoxicity of the test item was observed. An evaluation was made using data of all the six examined concentrations. No biologically relevant or statistically significant increase was observed at the evaluated concentrations. No significant dose response to the treatment was indicated by the linear trend analysis.
The experiments were performed using appropriate untreated, negative (solvent) and positive control samples in all cases. The spontaneous mutation frequency of the negative (solvent) controls was in the recommended range in each test. The positive controls gave the anticipated increases in mutation frequency over the controls. The plating efficiencies for the solvent controls at the end of the expression period were accepted in all assays. The evaluated concentration ranges were considered to be adequate. The number of test concentrations met the acceptance criteria. Therefore, all the validity criteria were fulfilled and the study was considered to be valid and to reflect the real potential of the test item to cause mutations in the cultured mouse cells used in this study.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
In vitro bacterial gene mutation: Key study: The Bacterial Reverse mutation Assay (Ames test) for the test substance Wasox-MMAC2 was performed according to OECD Guideline 471 and GLP. According to the results obtained in this study, the test substance is non-mutagenic in the Ames test to the Salmonella typhimurium strains TA97a, TA98, TA100, TA102 and TA1535 with and without metabolic activation up to 5000 µg/plate.
In vitro cytogenicity in mammalian cells: Key study: In vitro Mammalian Chromosome Aberration Test in human lymphocytes was performed according to OECD Guideline 473. Under study conditions, there was no relevant evidence that the test substance Wasox-MMAC2 did induce structural chromosomal aberrations in cultured human Iymphocytes, neither without nor with the use of a metabolic activation system.
In vitro gene mutation in mammalian cells: Read-across approach from experimental results on an analogue substance: Key study: An in vitro mammalian cell assay was performed in mouse lymphoma L5178Y TK+/- 3.7.2 C cells at the tk locus to test the potential of acetone oxime to cause gene mutation and/or chromosome damage. The test item was determined to be non mutagenic either in the presence or in the absence of metabolic activation system under the conditions of this Mouse Lymphoma Assay.
Justification for classification or non-classification
Based on the available information on genetic toxicity in vitro, the substance is considered to be negative, and therefore, the substance is not classified for mutagenicity according to CLP Regulation (EC) no. 1272/2008.
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