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EC number: 228-036-4 | CAS number: 6092-54-2
- 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
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:
- June 2015
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP / OECD guideline study.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 015
- Report date:
- 2015
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Hexyl chloroformate
- EC Number:
- 228-036-4
- EC Name:
- Hexyl chloroformate
- Cas Number:
- 6092-54-2
- Molecular formula:
- C7H13ClO2
- IUPAC Name:
- hexyl carbonochloridate
- Test material form:
- other: liquid
- Details on test material:
- - Name of test material (as cited in study report): N-hexyl chloroformate
- Physical state: liquid, colorless, clear
- Analytical purity: 97.3 area-%
- Stability under test conditions: guaranteed until 16 Oct 2015 as indicated by the sponsor
- Storage condition of test material: RT (protect against humidity, avoid temperatures >40°C)
Constituent 1
Method
- Target gene:
- Salmonella typhimurium: histidine
Escherichia coli WP2 uvrA: tryptophan
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:
- Phenobarbital/b-Naphthoflavone induced rat liver S9
- Test concentrations with justification for top dose:
- Standard plate test: 33; 100; 333; 1000; 2500; and 5000 µg/plate
Preincubation test: 10; 33; 100; 333; 1000; and 2500 μg/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: test substance insolube in water
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: With S9 mix: 2-aminoanthracene (2-AA). W/o S9 mix: N-methyl-N'-nitro-N-nitrosoguanidine (MNNG); 4-nitro-o-phenylenediamine (NOPD); 9-aminoacridine (AAC), 4-nitroquinoline-N-oxide (4-NQO).
- Details on test system and experimental conditions:
- 1. Standard plate test +/- S9 mix:
METHOD OF APPLICATION: in agar (plate incorporation) according to Ames et al. 1975, Maron et al. 1983
NUMBER OF REPLICATIONS: 3 test plates /dose or control
2. Preincubation Test +/- S9 mix (because no mutagenicity was observed in the Standard plate test)
METHOD OF APPLICATION: in medium, according to Yahagi et al. 1977 und Matsushima et al. 1980
NUMBER OF REPLICATIONS: 3 test plates /dose or control - Evaluation criteria:
- Acceptance criteria
Generally, the experiment was considered valid if the following criteria were met:
• The number of revertant colonies in the negative controls was within the range of the historical negative control data for each tester strain.
• The sterility controls revealed no indication of bacterial contamination.
• The positive control substances both with and without S9 mix induced a distinct increase in the number of revertant colonies within the range of the historical positive control data or above.
• Fresh bacterial culture containing approximately 10exp9 cells per mL were used.
Assessment criteria
The test substance was considered positive in this assay if the following criteria were met:
• A dose-related and reproducible increase in the number of revertant colonies, i.e. at least doubling (bacteria strains with high spontaneous mutation rate, like TA 98, TA 100 and E.coli WP2 uvrA) or tripling (bacteria strains with low spontaneous mutation rate, like TA 1535 and TA 1537) of the spontaneous mutation rate in at least one tester strain either without S9 mix or after adding a metabolizing system.
A test substance was generally considered non-mutagenic in this test if:
• The number of revertants for all tester strains were within the historical negative control data range under all experimental conditions in at least two experiments carried out independently of each other.
Results and discussion
Test results
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- >= 2500 μg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
No test substance precipitation was found with and without S9 mix.
COMPARISON WITH HISTORICAL CONTROL DATA:
The results of the negative as well as the positive controls performed in parallel corroborated the validity of this study, since the values fulfilled the acceptance criteria of this study.
In this study with and without S9 mix, the number of revertant colonies in the negative controls was within or marginally below the range of the historical negative control data for each tester strain.
In addition, the positive control substances both with and without S9 mix induced a significant increase in the number of revertant colonies within the range of the historical positive control data.
ADDITIONAL INFORMATION ON CYTOTOXICITY:
A bacteriotoxic effect (reduced his- or trp- background growth, decrease in the number of his+ or trp+ revertants) was observed in the standard plate test and in the preincubation assay under all test conditions from about 2500 μg/plate onward. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
Without S9 Mix
Standard Plate Test - S9 mix | Preicubation Assay - S9 mix | |||||||||
Strain | Test group | Dose (µg/plate) | Mean revertants per plate | Standard deviation | Factor | Reduced background growth | Mean revertants per plate | Standard deviation | Factor | Reduced background growth |
TA 1535 | DMSO | - | 13,7 | 3,5 | - | 12,3 | 2,1 | - | ||
Test item | 10 | - | - | - | 14,7 | 3,1 | 1,2 | |||
33 | 10,0 | 2,6 | 0,7 | 12,3 | 4,0 | 1,0 | ||||
100 | 11,0 | 1,0 | 0,8 | 16,7 | 8,4 | 1,4 | ||||
333 | 1,7 | 5,7 | 0,9 | 10,3 | 2,0 | 0,8 | ||||
1000 | 11,0 | 2,0 | 0,8 | 11,0 | 1,7 | 0,9 | ||||
2500 | 6,0 | 11,7 | 0,4 | x | 7,7 | 3,8 | 0,6 | x | ||
5000 | 0,0 | 0,0 | 0,0 | x | - | - | - | |||
MNNG | 5,0 | 5114,7 | 351,0 | 374,2 | 3214,0 | 84,9 | 260,6 | |||
TA 100 | DMSO | - | 94,0 | 1,7 | - | 81,7 | 1,5 | - | ||
Test item | 10 | - | - | - | 80,0 | 9,6 | 1,0 | |||
33 | 88,7 | 1,2 | 0,9 | 91,0 | 10,0 | 1,1 | ||||
100 | 94,0 | 12,3 | 1,0 | 84,3 | 8,1 | 1,0 | ||||
333 | 90,0 | 10,6 | 1,0 | 86,7 | 406,0 | 1,1 | ||||
1000 | 87,3 | 2,1 | 0,9 | 80,7 | 17,4 | 1,0 | ||||
2500 | 78,3 | 8,3 | 0,8 | x | 29,7 | 22,5 | 0,4 | x | ||
5000 | 0,0 | 0,0 | 0,0 | x | - | - | - | |||
MNNG | 5,0 | 4110,0 | 379,5 | 43,7 | 2186,3 | 90,2 | 26,8 | |||
TA 1537 | DMSO | - | 6,7 | 1,2 | - | 8,0 | 3,5 | - | ||
Test item | 10 | - | - | - | 8,3 | 4,5 | 1,0 | |||
33 | 6,7 | 0,6 | 1,0 | 9,7 | 5,5 | 1,2 | ||||
100 | 6,3 | 3,1 | 0,9 | 8,3 | 1,2 | 1,0 | ||||
333 | 8,0 | 2,0 | 1,2 | 10,3 | 5,0 | 1,3 | ||||
1000 | 8,7 | 3,1 | 1,3 | 6,0 | 2,6 | 0,8 | ||||
2500 | 7,0 | 4,0 | 1,0 | x | 3 | 1,0 | 0,4 | x | ||
5000 | 0,0 | 0,0 | 0,0 | x | - | - | - | |||
AAC | 100,0 | 943,0 | 193,8 | 141,5 | 473,3 | 46,0 | 59,2 | |||
TA 98 | DMSO | - | 14,3 | 1,5 | - | 22 | 3,6 | - | ||
Test item | 10 | - | - | - | 19,3 | 3,8 | 0,9 | |||
33 | 17,3 | 0,6 | 1,2 | 20 | 3,6 | 0,9 | ||||
100 | 19,7 | 2,5 | 1,4 | 19,3 | 4,5 | 0,9 | ||||
333 | 14,0 | 4,4 | 1,0 | 24 | 3,6 | 1,1 | ||||
1000 | 13,7 | 1,5 | 1,0 | 20 | 1,0 | 0,9 | ||||
2500 | 12,7 | 4,5 | 0,9 | x | 5 | 4,0 | 0,2 | x | ||
5000 | 0,0 | 0,0 | 0,0 | x | - | - | - | |||
NOPD | 10,0 | 447,0 | 8,5 | 31,2 | 324 | 35,8 | 14,7 | |||
E. coli | DMSO | - | 20,0 | 2,6 | - | 21,7 | 1,2 | - | ||
Test item | 10 | - | - | - | 22,7 | 7,1 | 1,0 | |||
33 | 18,7 | 6,1 | 0,9 | 20,0 | 1,0 | 0,9 | ||||
100 | 22,7 | 2,5 | 1,1 | 21,7 | 6,4 | 1,0 | ||||
333 | 16,7 | 7,4 | 0,8 | 17,7 | 4,9 | 0,8 | ||||
1000 | 19,3 | 5,9 | 1,0 | 17,7 | 4,2 | 0,8 | ||||
2500 | 18,0 | 3,5 | 0,9 | x | 12,0 | 4,6 | 0,6 | x | ||
5000 | 0,0 | 0,0 | 0,0 | x | - | - | - | |||
4-NQO | 5 | 1150,3 | 15,0 | 57,5 | 433,7 | 65,7 | 20,0 |
With S9 Mix
Standard Plate Test + S9 mix | Preincubation Assay + S9 mix | |||||||||
Strain | Test group | Dose (µg/plate) | Mean revertants per plate | Standard deviation | Factor | Reduced background growth | Mean revertants per plate | Standard deviation | Factor | Reduced background growth |
TA 1535 | DMSO | - | 10,0 | 1,0 | - | 16,0 | 2,0 | - | ||
10 | - | - | - | 17,7 | 2,5 | 1,1 | ||||
Test item | 33 | 11,0 | 5,0 | 1,1 | 11,3 | 5,1 | 0,7 | |||
100 | 12,7 | 3,2 | 1,3 | 13,3 | 3,5 | 0,8 | ||||
333 | 9,3 | 3,1 | 0,9 | 10,3 | 3,5 | 0,6 | ||||
1000 | 8,7 | 5,7 | 0,9 | 13,0 | 3,0 | 0,8 | ||||
2500 | 13,0 | 5,6 | 1,3 | x | 8,3 | 1,5 | 0,5 | x | ||
5000 | 0,0 | 0,0 | 0,0 | x | - | - | - | |||
2-AA | 2,5 | 176,3 | 5,5 | 27,6 | 215,7 | 26,4 | 13,5 | |||
TA 100 | DMSO | - | 84,0 | 11,8 | - | 87,7 | 2,9 | - | ||
10 | - | - | - | 84,7 | 2,5 | 1,0 | ||||
Test item | 33 | 90,3 | 5,7 | 1,1 | 84,7 | 1,5 | 1,0 | |||
100 | 92,0 | 22,1 | 1,1 | 96,7 | 10,4 | 1,1 | ||||
333 | 78,0 | 21,6 | 0,9 | 88,7 | 6,5 | 1,0 | ||||
1000 | 87,3 | 4,5 | 1 | 82,0 | 9,5 | 0,9 | ||||
2500 | 73,3 | 5,9 | 0,9 | x | 14,3 | 16,2 | 0,2 | x | ||
5000 | 0,0 | 0,0 | 0,0 | x | - | - | - | |||
2-AA | 2,5 | 1515,7 | 160,5 | 18,0 | 1460,7 | 75,5 | 16,7 | |||
TA 1537 | DMSO | - | 11,7 | 2,5 | - | 8,0 | 3,5 | - | ||
10 | - | - | - | 9,7 | 1,5 | 1,2 | ||||
Test item | 33 | 11,7 | 0,6 | 1,0 | 7,7 | 2,5 | 1,0 | |||
100 | 11,7 | 1,5 | 1,0 | 9,3 | 2,1 | 1,2 | ||||
333 | 16,0 | 3,5 | 1,4 | 10,3 | 1,5 | 1,3 | ||||
1000 | 9,0 | 1,7 | 0,8 | 8,0 | 3,0 | 1,0 | ||||
2500 | 6,3 | 3,2 | 0,5 | x | 3,3 | 2,1 | 0,4 | x | ||
5000 | 0,0 | 0,0 | 0,0 | x | - | - | - | |||
2-AA | 2,5 | 106,7 | 9,9 | 9,1 | 106,7 | 4,9 | 13,3 | |||
TA 98 | DMSO | - | 14,3 | 4,9 | - | 21,3 | 2,5 | - | ||
10 | - | - | - | 19,3 | 2,5 | 0,9 | ||||
Test item | 33 | 11,0 | 5,6 | 0,8 | 18,7 | 3,5 | 0,9 | |||
100 | 15,7 | 5,5 | 1,1 | 20,7 | 6,5 | 1,0 | ||||
333 | 11,7 | 1,5 | 0,8 | 20,0 | 1,7 | 0,9 | ||||
1000 | 9,7 | 1,5 | 0,7 | 21,0 | 1,7 | 1,0 | ||||
2500 | 9,7 | 2,5 | 0,7 | x | 2,7 | 1,5 | 0,1 | x | ||
5000 | 5,3 | 1,2 | 0,4 | x | - | - | - | |||
2-AA | 2,5 | 1192,7 | 328,9 | 83,2 | 1252,7 | 75,5 | 57,7 | |||
E. coli | DMSO | - | 16,7 | 5,0 | - | 20,3 | 6,4 | - | ||
10 | - | - | - | 19,3 | 2,9 | 1,0 | ||||
Test item | 33 | 19,0 | 4,4 | 1,1 | 18,7 | 5,5 | 0,9 | |||
100 | 15,7 | 4,0 | 0,9 | 21,6 | 5,5 | 1,1 | ||||
333 | 17,0 | 6,6 | 1,0 | 20,7 | 7,2 | 1,0 | ||||
1000 | 23,3 | 3,8 | 1,4 | 17,3 | 6,0 | 0,9 | ||||
2500 | 18,3 | 0,6 | 1,1 | x | 11,7 | 2,5 | 0,6 | x | ||
5000 | 0,0 | 0,0 | 0,0 | x | - | - | - | |||
2-AA | 60 | 91,3 | 16,2 | 5,5 | 132,0 | 28,5 | 6,5 |
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative
In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used. - Executive summary:
The test substance N-Hexyl chloroformate was tested for its mutagenic potential based on the ability to induce point mutations in selected loci of several bacterial strains (S. typhimurium TA 1535, TA 100, TA 1537, TA 98 and E. coli WP2 uvrA) in a reverse mutation assay (GLP OECD471 guideline study; BASF SE, 2015).
Standard plate test (SPT) and preincubation test (PIT) both with and without metabolic activation (liver S9 mix from induced rats) were performed with concentration ranges of 33 μg - 5000 μg/plate (SPT) and 10 μg - 2500 μg/plate (PIT).
No precipitation of the test substance was found with and without S9 mix. A bacteriotoxic effect was observed from about 2500 μg/plate onward. A relevant increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the preincubation test either without S9 mix or after the addition of a metabolizing system.
Thus, under the experimental conditions of this study, the test substance N-Hexyl chloroformate is not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay in the absence and the presence of metabolic activation.
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