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EC number: 225-791-1 | CAS number: 5080-22-8
- 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 vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 3 - 23 January 2012
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: The study was conducted according to test guidelines and in accordance with GLP
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 012
- Report date:
- 2012
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
- Version / remarks:
- METI Act No. 117, Ministerial Ordinance No. 1-3 (2004)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- N-isopropylhydroxylamine
- EC Number:
- 225-791-1
- EC Name:
- N-isopropylhydroxylamine
- Cas Number:
- 5080-22-8
- Molecular formula:
- C3H9NO
- IUPAC Name:
- N-(propan-2-yl)hydroxylamine
- Details on test material:
- IPHA (Isopropylhydroxylamine, XU-18770.00) with a 99.7% purity by gas chromatography was used with identification confirmed by nuclear magnetic resonance and mass spectrometry.
Constituent 1
Method
- Target gene:
- Salmonella typhimurium histidine auxotrophs and and Escherichia coli WP2 uvrA
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254-induced rat liver S9 was used as the metabolic activation system.
- Test concentrations with justification for top dose:
- 1.5, 5.0, 15, 50, 150, 500, 1500 and 5000 ug/plate for all strains with and without S9 in introductory assay.
50,150,500,1500 and 5000 ug/plate for all strains with and without S9 in confirmatory assay. - Vehicle / solvent:
- water
Controls
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 2-nitrofluorene (TA98), sodium azide (TA100 and TA1535), 9-Aminoacridine (TA1537) and methyl methanesulfona (WP2uvrA) without S9 and 2-aminoanthracene (all strains) with S9
- Details on test system and experimental conditions:
- Initial Toxicity-Mutation Assay
The initial toxicity-mutation assay was used to establish the dose-range for the confirmatory mutagenicity assay and to provide a preliminary mutagenicity evaluation. Vehicle control, positive controls and a minimum of eight dose levels of the test article were plated, two plates per dose, with overnight cultures of TA98, TA100, TA1535, TA1537 and WP2 uvrA on selective minimal agar in the presence and absence of Aroclor-induced rat liver S9.
Confirmatory Mutagenicity Assay
The confirmatory mutagenicity assay was used to evaluate and confirm the mutagenic potential of the test article. A minimum of five dose levels of test article along with appropriate vehicle control and positive controls were plated with overnight cultures of TA98, TA100, TA1535, TA1537 and WP2 uvrA on selective minimal agar in the presence and absence of Aroclor-induced rat liver S9. All dose levels of test article, vehicle control and positive controls were plated in triplicate.
Plating and Scoring Procedures
The test system was exposed to the test article via the preincubation methodology described by Yahagi et al. (1977). On the day of its use, minimal top agar, containing 0.8 % agar (W/V) and 0.5 % NaCl (W/V), was melted and supplemented with L-histidine, D-biotin and L-tryptophan solution to a final concentration of 50 μM each. Top agar not used with S9 or Sham mix was supplemented with 25 mL of water for each 100 mL of minimal top agar. For the preparation of media and reagents, all references to water imply deionized water. Bottom agar was Vogel-Bonner minimal medium E (Vogel and Bonner, 1956) containing 1.5 % (W/V) agar. Nutrient bottom agar was Vogel-Bonner minimal medium E containing 1.5 % (W/V) agar and supplemented with 2.5 % (W/V) Oxoid Nutrient Broth No. 2 (dry powder). Nutrient Broth was Vogel-Bonner salt solution supplemented with 2.5 % (W/V) Oxoid Nutrient Broth No. 2 (dry powder).
Each plate was labeled with a code system that identified the test article, test phase, concentration, tester strain and activation, as described in detail in BioReliance's Standard Operating Procedures.
One-half (0.5) milliliter of S9 or sham mix, 100 μL of tester strain (cells seeded) and 100 μL of vehicle or test article dilution were added to 13 X 100 mm glass culture tubes pre-heated to 37±2°C. After vortexing, these mixtures were incubated with shaking for 20±2 minutes at 37±2°C. Following the preincubation, 2.0 mL of selective top agar was added to each tube and the mixture was vortexed and overlaid onto the surface of 25 mL of minimal bottom agar. When plating the positive controls, the test article aliquot was replaced by a 50 μL aliquot of appropriate positive control. After the overlay had solidified, the plates were inverted and incubated for approximately 48 to 72 hours at 37±2°C. Plates that were not counted immediately following the incubation period were stored at 2-8°C until colony counting could be conducted. - Evaluation criteria:
- The condition of the bacterial background lawn was evaluated for evidence of test article toxicity by using a dissecting microscope. Precipitate was evaluated by visual examination without magnification. Toxicity and degree of precipitation were scored relative to the vehicle control plate using the codes shown in the following table.
Revertant colonies for a given tester strain and activation condition, except for positive controls, were counted either entirely by automated colony counter or entirely by hand unless the plate exhibited toxicity.
For each replicate plating, the mean and standard deviation of the number of revertants per plate were calculated and are reported.
For the test article to be evaluated positive, it must cause a reproducible, concentration-related increase in the mean revertants per plate of at least one tester strain over a minimum of two increasing concentrations of test article. Data sets for tester strains TA98, TA1535, TA1537 and WP2 uvrA were judged positive if the increase in mean revertants at the peak of the response was greater than or equal to 3.0-times the mean vehicle control value. Data sets for tester strain TA100 were judged positive if the increase in mean revertants at the peak of the response was greater than or equal to 2.0-times the mean vehicle control value.
An equivocal response is a biologically relevant increase in a revertant count that partially meets the criteria for evaluation as positive. This could be a dose-responsive increase that does not achieve the respective threshold cited above or a non-dose responsive increase that is equal to or greater than the respective threshold cited. A response was evaluated as negative, if it was neither positive nor equivocal. - Statistics:
- Statistical analysis was not conducted.
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Solubility Test
Water was selected as the solvent of choice based on the Sponsor’s request, solubility of the test article and compatibility with the target cells. The test article formed a clear solution in water at approximately 50 mg/mL, the maximum concentration tested in the solubility test.
Sterility Results
No contaminant colonies were observed on the sterility plates for the vehicle control, the test article dilutions and the S9 and Sham mixes.
Initial Toxicity-Mutation Assay
In Experiment B1 (Initial Toxicity-Mutation Assay), the maximum concentration tested was 5000 μg per plate, which is the maximum concentration recommended by test guidelines. This concentration was achieved using a concentration of 50 mg/mL and a plating aliquot of 100 μL. The concentrations tested were 1.5, 5.0, 15, 50, 150, 500, 1500 and 5000 μg per plate. No positive mutagenic responses were observed with any of the tester strains in either the presence or absence of S9 activation. Neither precipitate nor background toxicity was observed. Based on the findings of the initial toxicity-mutation assay, the maximum dose plated in the confirmatory mutagenicity assay was 5000 μg per plate.
Confirmatory Mutagenicity Assay
In Experiment B2 (Confirmatory Mutagenicity Assay), no positive mutagenic responses were observed with any of the tester strains in either the presence or absence of S9 activation. The dose levels tested were 50, 150, 500, 1500 and 5000 μg per plate. Neither precipitate nor background lawn toxicity was observed.
Dosing Formulation Analysis
Dosing formulations were analyzed by the Sponsor. Concentration analysis indicates that the actual mean concentrations of the analyzed dose levels were between 95.7 and 109.5% of their respective targets with a relative percent difference (RPD) of < 20%. This indicates that the regulatory-required top dose level was achieved and the results support the validity of the study conclusion. No test article was detected in the vehicle control sample.
Stability analysis was performed on the highest and lowest dose concentrations in the initial toxicity-mutation assay, and the results of the analysis for the lowest dose concentration were outside the acceptable range of 85 to 115% of dose (start). The stability analysis was repeated with samples from the confirmatory mutagenicity assay at concentrations ranging from 0.015 to 50 mg/mL. The results of the analysis on the formulations from the confirmatory assay found the test article to be stable in water at room temperature for the period of dosing.
Any other information on results incl. tables
Table 1
Initial Toxicity-Mutation Assay without S9 activation
Mean revertants +St. Dev./plate | ||||||
Article | Dose level per plate | TA98 | TA100 | TA1535 | TA1537 | WP2 uvrA |
IPHA |
5000 ug | 16 + 1 | 126 + 6 | 10 + 2 | 6 + 3 | 34 + 0 |
1500 ug | 16 + 5 | 118 + 0 | 8 + 1 | 3 + 2 | 33 + 0 | |
500 ug | 16 + 5 | 92 + 19 | 17 + 3 | 3 + 3 | 24 + 1 | |
150 ug | 17 + 3 | 106 + 6 | 8 + 1 | 7 + 3 | 30 + 4 | |
50 ug | 14 + 4 | 99 + 8 | 8 + 6 | 3 + 3 | 35 + 2 | |
15 ug | 12 + 6 | 116 + 13 | 7 + 2 | 4 + 1 | 32 + 1 | |
5.0 ug | 14 + 4 | 89 + 6 | 8 + 1 | 6 + 4 | 35 + 4 | |
1.5 ug | 14 + 3 | 108 + 10 | 8 + 1 | 3 + 0 | 32 + 7 | |
Water | 100 uL | 12 + 1 | 93 + 1 | 10 + 2 | 5 + 3 | 44 + 3 |
2 -Nitrofluorene | 1.0 ug | 549 + 34 | ND | ND | ND | ND |
Sodium Azide | 1.0 ug | ND | 684 + 69 | 417 + 3 | ND | ND |
9 -Aminoacridine | 75 ug | ND | ND | ND | 542 + 20 |
ND |
Methyl Methanesulfonate | 1000 ug | ND | ND | ND | ND | 392 + 65 |
ND = No Data
Table 2
Initial Toxicity-Mutation Assay with S9 activation
Mean revertants +St. Dev./plate | ||||||
Article | Dose level per plate | TA98 | TA100 | TA1535 | TA1537 | WP2 uvrA |
IPHA |
5000 ug | 17 + 6 | 124 + 4 | 8 + 0 | 5 + 5 | 52 + 9 |
1500 ug | 15 + 4 | 129 + 13 | 13 + 1 | 8 + 1 | 34 + 4 | |
500 ug | 10 + 1 | 110 + 11 | 15 + 2 | 4 + 0 | 26 + 12 | |
150 ug | 12 + 4 | 106 + NA | 9 + 3 | 2 + 1 | 35 + 2 | |
50 ug | 13 + 3 | 101 + 4 | 11 + 8 | 1 + 0 | 29 + 6 | |
15 ug | 18 + 8 | 106 + 12 | 12 + 1 | 5+ 3 | 35 + 4 | |
5.0 ug | 8 + 1 | 128 + 8 | 11 + 2 | 8+ 5 | 37 + 1 | |
1.5 ug | 11 + 6 | 100 + 14 | 9 + 5 | 4+ 1 | 34 + 6 | |
Water | 100 uL | 12 + 6 | 106 + 28 | 8 + 1 | 6 + 2 | 31 + 3 |
2 -Aminoanthracene | 1.0 ug | 129 + 6 | ND | 55 + 4 | 41 + 6 | ND |
2 -Aminoanthracene | 2.0 ug | ND | 446 + 62 | ND | ND | ND |
2 -Aminoanthracene | 15 ug | ND | ND | ND | ND |
455 + 34 |
ND = No Data
Table 3
Confirmatory Mutagenicity Assay without S9 activation
Mean revertants +St. Dev./plate | ||||||
Article | Dose level per plate | TA98 | TA100 | TA1535 | TA1537 | WP2 uvrA |
IPHA |
5000 ug | 14 + 5 | 141 + 18 | 13 + 2 | 4 + 2 | 19 + 7 |
1500 ug | 16 + 6 | 111 + 8 | 9 + 2 | 5 + 4 | 17 + 7 | |
500 ug | 10 + 2 | 107 + 12 | 11 + 1 | 4 + 3 | 22 + 7 | |
150 ug | 12 + 5 | 110 + 8 | 12 + 4 | 4 + 2 | 14 + 3 | |
50 ug | 9 + 3 | 106 + 3 | 11 + 1 | 6 + 3 | 16 + 6 | |
Water | 100 uL | 12 + 2 | 125 + 13 | 15 + 4 | 5 + 1 | 16 + 5 |
2 -Nitrofluorene | 1.0 ug | 451 + 114 | ND | ND | ND | ND |
Sodium Azide | 1.0 ug | ND | 650 + 31 | 530 + 27 | ND | ND |
9 -Aminoacridine | 75 ug | ND | ND | ND | 305 + 70 |
ND |
Methyl Methanesulfonate | 1000 ug | ND | ND | ND | ND | 299 + 32 |
ND = No Data
Table 4
Confirmatory Mutagenicity Assay with S9 activation
Mean revertants +St. Dev./plate | ||||||
Article | Dose level per plate | TA98 | TA100 | TA1535 | TA1537 | WP2 uvrA |
IPHA |
5000 ug | 15 + 6 | 151 + 6 | 6 + 2 | 8 + 4 | 23 + 5 |
1500 ug | 15 + 6 | 120 + 10 | 7 + 2 | 6 + 3 | 17 + 4 | |
500 ug | 13 + 6 | 138 + 9 | 12 + 1 | 5 + 1 | 24 + 8 | |
150 ug | 14 + 3 | 117 + 8 | 6 + 3 | 9 + 1 | 25 + 3 | |
50 ug | 8 + 2 | 138 + 25 | 11 + 3 | 4 + 1 | 18 + 2 | |
Water | 100 uL | 10 + 9 | 149 + 22 | 13 + 4 | 7 + 5 | 20 + 4 |
2 -Aminoanthracene | 1.0 ug | 540 + 85 | ND | 58 + 18 | 80 + 6 | ND |
2- Aminoanthracene | 2.0 ug | ND | 857 + 79 | ND | ND | ND |
2 -Aminoanthracene | 15 ug | ND | ND | ND | ND |
98 + 5 |
ND = No Data
Applicant's summary and conclusion
- Conclusions:
- All criteria for a valid study were met as described in the protocol. The results of the Bacterial Reverse Mutation Assay indicate that, under the conditions of this study, IPHA (Isopropylhydroxylamine) was negative (non-mutagenic) both in the presence and absence of
Aroclor-induced rat liver S9. - Executive summary:
The test article, IPHA (Isopropylhydroxylamine), was tested in the Bacterial Reverse Mutation Assay using Salmonella typhimurium tester strains TA98, TA100, TA1535 and TA1537 and Escherichia coli tester strain WP2 uvrA in the presence and absence of Aroclor-induced rat liver S9. The assay was performed in two phases using the preincubation method. The first phase, the initial
toxicity-mutation assay, was used to establish the dose-range for the confirmatory mutagenicity assay and to provide a preliminary mutagenicity evaluation. The second phase, the confirmatory mutagenicity assay, was used to evaluate and confirm the mutagenic potential of the test article.
Water was selected as the solvent of choice based on the Sponsor’s request, solubility of the test article and compatibility with the target cells. The test article formed a clear solution in water at approximately 50 mg/mL, the maximum concentration tested in the solubility test.
In the initial toxicity-mutation assay, the maximum concentration tested was 5000 μg per plate, which is the maximum concentration recommended by test guidelines. This concentration was achieved using a concentration of 50 mg/mL and a plating aliquot of 100 μL. The concentrations tested were 1.5, 5.0, 15, 50, 150, 500, 1500 and 5000 μg per plate. No positive mutagenic responses
were observed with any of the tester strains in either the presence or absence of S9 activation. Neither precipitate nor background lawn toxicity was observed. Based on the findings of the initial toxicity-mutation assay, the maximum dose plated in the confirmatory mutagenicity assay was 5000 μg per plate.
In the confirmatory mutagenicity assay, no positive mutagenic responses were observed with any of the tester strains in either the presence or absence of S9 activation. The dose levels tested were 50, 150, 500, 1500 and 5000 μg per plate. Neither precipitate nor background lawn toxicity was observed.
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