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EC number: 250-610-8 | CAS number: 31394-54-4
- 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
All read across genetic toxicity tests listed below had negative results for Isoheptane.
Genetic Toxicity in vitro – Bacterial reverse mutation assay (OECD 471)
Genetic Toxicity in vitro – Mammalian chromosome aberration test (OECD 473)
Genetic Toxicity in vitro – Mammalian cell gene mutation assay (OECD 476)
Genetic Toxicity in vitro – Sister chromatid exchange assay in mammalian cells (OECD 479)
Genetic Toxicity in vitro – Unscheduled DNA synthesis in mammalian cells (OECD 482)
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- 1982-1983
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline study with acceptable restrictions
- Justification for type of information:
- A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Principles of method if other than guideline:
- No specific method or guideline was noted; similar to OECD guideline 471; limited documentation
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- His-operon (Salmonellla), Trp-operon (E. coli)
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Species / strain / cell type:
- S. typhimurium TA 1538
- Species / strain / cell type:
- E. coli WP2
- Species / strain / cell type:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix of livers from Aroclor1254-pretreated rats
- Test concentrations with justification for top dose:
- max. conc. tested: 250 µg/mL
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Tween80/ethanol
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: benzo[a]pyrene in DMSO, 4-nitroquinoline-N-oxide in DMSO, sodium azide, neutral red, potassium dichromate
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: preincubation
DURATION
- Preincubation period: no data
- Exposure duration: not applicable, preincubation method
- Expression time (cells in growth medium): 48-72 hours
DETERMINATION OF CYTOTOXICITY
- Method: other: toxicity screening test - Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- 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:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
The purpose of this study was to determine the mutagenicity of the test substance Normal-Heptane. A reverse mutation assay was done using S. typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100, and E. coli strains WP2 and WP2 uvr A. The strains were exposed to concentrations of 0, 3.91, 7.81, 15.6, 31.3, 62.5, 125, and 250 ug/mL for 48 -72 hrs both with and without metabolic activation. The number of revertant colonies was then counted.
No significant increases in the ratio of mutations over controls was seen. The test substance is not mutagenic in either the presence or absence of metabolic activation. - Executive summary:
The purpose of this study was to determine the mutagenicity of the test substance Normal-Heptane. A reverse mutation assay was done using S. typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100, and E. coli strains WP2 and WP2 uvr A. The strains were exposed to concentrations of 0, 3.91, 7.81, 15.6, 31.3, 62.5, 125, and 250 ug/mL for 48 -72 hrs both with and without metabolic activation. The number of revertant colonies was then counted.
No significant increases in the ratio of mutations over controls was seen. The test substance is not mutagenic in either the presence or absence of metabolic activation.
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Study meets generally accepted scientific principles, acceptable for assessment.
- Justification for type of information:
- A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- - no A-T base pair tester strains included (e.g. TA102 or E.coli); limited documentation
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Deviations:
- yes
- Remarks:
- - no A-T base pair tester strains included (e.g. TA102 or E.coli); limited documentation
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his-operon
- Species / strain / cell type:
- S. typhimurium, other: TA 98, TA100, TA1535, TA1537, TA1538
- Details on mammalian cell type (if applicable):
- no data
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 liver microsomes from male Sprague Dawley rats treated with a single intraperitoneal injection of Aroclor 1254 5-days before sacrifice.
- Test concentrations with justification for top dose:
- 20, 60, 560, 1670, and 5000 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: N-methl-N'-nitro-N-nitrosoguanidine
- Remarks:
- positive control for TA100 and TA1535 (5 µg/plate)
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- 2-nitrofluorene
- Remarks:
- positive control for TA98 and TA1538 (50 µg/plate) Migrated to IUCLID6: in absence of S9-mix
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- positive control for TA1537 (75 µg/plate)
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 5000 µg/plate
- Vehicle 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:
- cytotoxicity
- Remarks:
- 5000 µg/plate
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 5000 µg/plate
- Vehicle 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:
- cytotoxicity
- Remarks:
- 5000 µg/plate
- Vehicle 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:
- cytotoxicity
- Remarks:
- 5000 µg/plate
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: strain/cell type: TA 98, TA100, TA1535, TA1537, TA1538
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
Under the test conditions, the experimental compound, Isooctane, did not exhibit a positive response and is, therefore, considered not be mutagenic in this system. - Executive summary:
Under the test conditions, the experimental compound, Isooctane, did not exhibit a positive response and is, therefore, considered not be mutagenic in this system.
- 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:
- key study
- Study period:
- 1982-1983
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline study with acceptable restrictions
- Justification for type of information:
- A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Principles of method if other than guideline:
- No guideline stated; similar to OECD guideline 473; rat hepatocytes with intrinsic metabolic activity, therefore no metabolic activation system added, only 100 cells from each culture analyzed.
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- not applicable
- Species / strain / cell type:
- hepatocytes: rat liver (RL4) cells
- Details on mammalian cell type (if applicable):
- The rat liver cell line, RL4, an epithelial-type cell line, was derived in this laboratory following the procedure described by Williams et al. (1971). RL4 was initiated in 1978 from a 10-day-old Wistar rat as described in Dean and Hodson-Walker (1979).
- Metabolic activation:
- not applicable
- Metabolic activation system:
- not necessary, liver cells
- Test concentrations with justification for top dose:
- 2.5, 5, 10 µg/ml
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Tween 80/ethanol
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 7,12-dimethylbenzanthracene in DMSO, ethyl methanesulphonate, cyclophosphamide
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 22 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 22 hours + 2 hours fixation
SPINDLE INHIBITOR (cytogenetic assays): colcemid
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: no data
NUMBER OF CELLS EVALUATED: 100 cells from each culture - Evaluation criteria:
- Significant or dose-related increase in chromosome damage.
- Key result
- Species / strain:
- hepatocytes: Rat liver (RL4) cells
- Metabolic activation:
- not applicable
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
This study examined the potential for the test substance Heptane to cause chromosomal aberrations in rat liver RL4 cells. Cells were exposed to concentrations of 0, 2.5, 5, and 10 ug/ml of test substance, and then examined for chromosomal aberrations including polyploidy, chromatid gaps, and chromatid exchanges. The positive control substance was 7,12 -dimethylbenzanthracene. A significant increase in chromatid gaps (7) was seen at the 10 ug/ml concentration. Though greater than the solvent control frequency, this was not accompanied by an increase in any other aberrations and is not considered to be treatment related. Under the conditions of this study, the test material was not clastogenic. - Executive summary:
This study examined the potential for the test substance Normal-Heptane to cause chromosomal aberrations in rat liver RL4 cells. Cells were exposed to concentrations of 0, 2.5, 5, and 10 ug/ml of test substance, and then examined for chromosomal aberrations including polyploidy, chromatid gaps, and chromatid exchanges. The positive control substance was 7,12 -dimethylbenzanthracene. A significant increase in chromatid gaps (7) was seen at the 10 ug/ml concentration. Though greater than the solvent control frequency, this was not accompanied by an increase in any other aberrations and is not considered to be treatment related. Under the conditions of this study, the test material was not clastogenic.
- 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:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Acceptable, well documented publication which meets basic scientific principles
- Justification for type of information:
- A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- yes
- Remarks:
- - using microtiter plates
- GLP compliance:
- not specified
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- thymidine kinase
- Species / strain / cell type:
- human lymphoblastoid cells (TK6)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPMI 1640 medium supplemented with 15% heat-inactivated horse serum
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- with Aroclor induced-rat liver homogenate
- Test concentrations with justification for top dose:
- 5 % v/v TMP in DMEM medium, administered undiluted or as 50 % (1:1 saturated to normal medium)
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMEM medium
- Untreated negative controls:
- yes
- Remarks:
- with and without S9
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMEM medium
- Positive controls:
- yes
- Remarks:
- for S9 activated cultures
- Positive control substance:
- benzo(a)pyrene
- Remarks:
- Migrated to IUCLID6: 15 µM
- Untreated negative controls:
- yes
- Remarks:
- with and without S9
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMEM medium
- Positive controls:
- yes
- Remarks:
- for non-activated cultures
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- Migrated to IUCLID6: 0.2 mM
- Details on test system and experimental conditions:
- see "any other information on materials and methods"
- Key result
- Species / strain:
- human lymphoblastoid cells (TK6)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- TMP did not induce significant increases in the mutation frequency at the thymidine kinase locus.
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Cell survival in TMP-saturated medium with and without metabolic activation was greater than 50-60 %.
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- The elevated mutation frequencies of positive control compounds were as expected.
ADDITIONAL INFORMATION ON CYTOTOXICITY:
Cell survival in positive control treated cultures was 40 or 30% for benzo(a)pyrene and EMS, respectively - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
Based on the study design there is no incidence of increased genetic toxicity caused by the test substance. - Executive summary:
Based on the study design there is no incidence of increased genetic toxicity caused by the test substance.
Referenceopen allclose all
The addition of heptane at amounts up to 250 µg per mL to cultures of Escherichia coli WP2 and WP2 uvr A, Salmonella typhimurium TA 1535, TA 1537, TA 1538, TA 98, and TA 100 did not lead to an increase in the reverse gene mutation frequency in any of these strains, either in the presence or in the absence of rat liver S9 fraction.
Table: Relative Reverse Mutation Rate – E. coli
Concentration (µg/ml) |
E. coliWP2 Assay 1 |
E. coliWP2 Assay 2 |
E. coliWP2 Assay 3 |
E. coliWP2 uvr A Assay 1 |
E. coliWP2 uvr A Assay 2 |
Without S9 |
|||||
3.91 |
- |
1.6 |
0.9 |
- |
0.9 |
7.81 |
- |
2.3 |
1.0 |
- |
0.9 |
15.6 |
1.2 |
1.3 |
0.8 |
0.5 |
0.8 |
31.3 |
1.1 |
1.4 |
0.7 |
0.8 |
0.6 |
62.5 |
1.0 |
1.0 |
0.8 |
0.9 |
0.5 |
125 |
0.7 |
0.6 |
1.2 |
0.3 |
0.5 |
250 |
0.5 |
0.8 |
1.0 |
0.2 |
0.5 |
4-nitroquinoline-N-oxide |
31.7 |
7.7 |
4.8 |
1.9 |
6.3 |
With S9 |
|||||
3.91 |
- |
1.5 |
0.9 |
- |
1.0 |
7.81 |
- |
2.4 |
0.9 |
- |
1.4 |
15.6 |
0.7 |
3.1 |
1.0 |
1.8 |
0.9 |
31.3 |
0.7 |
2.8 |
0.8 |
0.8 |
0.9 |
62.5 |
1.0 |
2.4 |
0.6 |
1.0 |
0.9 |
125 |
0.9 |
2.2 |
1.1 |
2.0 |
0.9 |
250 |
0.7 |
1.3 |
0.9 |
- |
0.8 |
4-nitroquinoline-N-oxide |
1.0 |
3.9 |
0.7 |
9.0 |
19.7 |
Table: Relative Mutation Rate – S. typhimurium TA 1535, TA 1537, TA 1538
Concentration (µg/ml) |
TA 1535 Assay 1 |
TA 1535 Assay 2 |
TA 1537 Assay 1 |
TA 1537 Assay 2 |
TA 1538 Assay 1 |
TA 1538 Assay 2 |
TA 1538 Assay 3 |
Without S9 |
|||||||
3.91 |
- |
- |
- |
- |
- |
- |
- |
7.81 |
- |
1.2 |
- |
0.7 |
- |
0.8 |
1.0 |
15.6 |
1.6 |
1.2 |
1.3 |
0 |
0.5 |
1.0 |
1.0 |
31.3 |
0.1 |
0.6 |
0.4 |
0 |
0 |
0.4 |
0.7 |
62.5 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
125 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
250 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
Sodium azide 1.7 µg |
48.0 |
73.1 |
- |
- |
- |
- |
- |
Benzo(a)-pyrene 6.7 µg |
- |
- |
- |
- |
1.4 |
1.1 |
0.9 |
Neutral red 6.7 µg |
- |
- |
1.5 |
1.6 |
- |
- |
- |
With S9 |
|||||||
3.91 |
- |
- |
- |
- |
- |
- |
- |
7.81 |
- |
0.9 |
- |
0.7 |
- |
- |
0.8 |
15.6 |
2.1 |
1.0 |
2.4 |
0.7 |
1.5 |
- |
1.0 |
31.3 |
1.4 |
0.9 |
1.3 |
0.9 |
2.1 |
- |
0.7 |
62.5 |
1.7 |
0.6 |
1.8 |
0.9 |
1.8 |
- |
1.0 |
125 |
1.7 |
1.3 |
2.1 |
0.6 |
1.2 |
- |
1.1 |
250 |
1.1 |
0.7 |
2.3 |
0.8 |
1.3 |
- |
0.9 |
Sodium azide 1.7 µg |
19.3 |
78.3 |
- |
- |
- |
- |
- |
Benzo(a)-pyrene 6.7 µg |
- |
- |
- |
- |
2.4 |
- |
10.6 |
Neutral red 6.7 µg |
- |
- |
3.0 |
6.0 |
- |
- |
- |
Table: Relative Mutation Rate – S. typhimurium TA 98, TA 100
Concentration (µg/ml) |
TA 98 Assay 1 |
TA 98 Assay 2 |
TA 100 Assay 1 |
TA 100 Assay 2 |
Without S9 |
||||
3.91 |
- |
0.5 |
- |
1.1 |
7.81 |
- |
0.1 |
- |
1.0 |
15.6 |
0.6 |
0 |
1.1 |
0.8 |
31.3 |
0 |
0 |
1.1 |
0.1 |
62.5 |
0 |
0 |
1.0 |
0 |
125 |
0 |
0 |
1.2 |
0 |
250 |
0 |
0 |
0.6 |
0 |
Benzo(a)-pyrene 6.7 µg |
0.8 |
1.2 |
1.6 |
1.0 |
With S9 |
||||
3.91 |
- |
0.8 |
- |
1.0 |
7.81 |
- |
1.1 |
- |
0.9 |
15.6 |
0.9 |
1.1 |
0.9 |
1.0 |
31.3 |
0.9 |
1.0 |
0.8 |
1.1 |
62.5 |
0.8 |
0.9 |
0.7 |
0.9 |
125 |
0.7 |
0.8 |
0.9 |
1.2 |
250 |
0.4 |
0.5 |
0.9 |
0.9 |
Benzo(a)-pyrene 6.7 µg |
12.6 |
4.7 |
2.8 |
5.4 |
TA98: Revertants Test
Test substance | Test 1 (without S9) | Test 2 (without S9) | Test 1 (with S9) | Test 2 (with S9) |
solvent control | 22 -26 | 22 -26 | 22 -26 | 18 -20 |
2 -Nitrofluorene | 1508 -1637 | 2131 -2282 | - | - |
2 -Aminoanthracene | - | - | 2213 -2247 | 1913 -2020 |
7 µg/plate isooctane | - | 15 -27 | - | 24 -46 |
20 µg/plate isooctane | 140 -191 | 19 -23 | 145 -206 | 29 -31 |
60 µg/plate isooctane | 112 -124 | 19 -26 | 64 -117 | 24 -33 |
190 µg/plate isooctane | 37 -55 | 19 -24 | 17 -28 | 12 -25 |
Retest (test 2) of TA98 without and with S9 at dose levels for 7, 20 and 60 µg/plate evidenced no difference from solvent controls.
Observed 5 -10 fold increase of TA98 revertants without and with S9 at 20 and 60 µg/plate in initial test was not confirmed in retest of TA98 without and with S9 at dose levels 7, 20 and 60 µg/plate.
In all experiments, no increase in revertants for other frame shift strains (TA1537 and TA 1538) was observed. Therefore the in test 1 observed genotoxic effects of isooctane in strain TA 98 were rather false positive.
There was no significant or dose-related increase of chromosome damage in any of the cultures exposed.
Table: Metaphase chromosome analysis of RL4cells
0 µg/ml |
2.5 µg/ml |
5.0 µg/ml |
10.0 µg/ml |
1.0 µg/ml 7,12-dimethylbenzanthracene |
|
Polyploidy (% cells) |
3.7 |
2.0 |
2.7 |
1.3 |
1.5 |
Chromatid gaps (% cells) |
1.3 |
1.7 |
1.0 |
2.3 |
6.0 |
Multiple aberrations (% cells) |
- |
- |
- |
- |
0.5 |
Severe damage (% cells) |
- |
- |
- |
- |
0.5 |
Chromatid aberrations (% cells) |
0.3 |
- |
- |
- |
4.0 |
Chromosome aberrations (frequency per cell) |
- |
- |
- |
- |
- |
Chromatid gaps (frequency per cell) |
0.013 |
0.017 |
0.010 |
0.027 |
0.085 |
Chromatid breaks (frequency per cell) |
0.003 |
- |
- |
- |
0.005 |
Chromatid exchange (frequency per cell) |
- |
- |
- |
- |
0.05 |
Chromosome breaks (frequency per cell) |
- |
- |
- |
- |
0.005 |
Chromosome exchange (frequency per cell) |
- |
- |
- |
- |
- |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
Read across genetic toxicity tests listed below has negative results for Isoheptane.
Genetic Toxicity in vivo – rodent dominant lethal assay (OECD 478)
Genetic Toxicity in vivo – unscheduled DNA synthesis (OECD 486)
Link to relevant study records
- Endpoint:
- in vivo mammalian cell study: DNA damage and/or repair
- Remarks:
- Type of genotoxicity: DNA damage and/or repair
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline study with acceptable restrictions.
- Justification for type of information:
- A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 486 (Unscheduled DNA Synthesis (UDS) Test with Mammalian Liver Cells in vivo)
- Deviations:
- yes
- Remarks:
- - limited documentation
- GLP compliance:
- not specified
- Type of assay:
- unscheduled DNA synthesis
- Species:
- rat
- Strain:
- Fischer 344
- Sex:
- male
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Breeding Laboratories (Kingston, N.Y.)
- Weight at study initiation: 200-300 g
- Assigned to test groups randomly: yes
- Fasting period before study: no
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
ENVIRONMENTAL CONDITIONS
- Temperature (°C): controlled
- Humidity (%): controlled
- Photoperiod (hrs dark / hrs light): 12/12 - Route of administration:
- oral: gavage
- Vehicle:
- - Vehicle(s)/solvent(s) used: none
- Duration of treatment / exposure:
- one single dose
- Frequency of treatment:
- sinlge treatment
- Post exposure period:
- 2, 12, 24 and 48 hours after dosing
- Remarks:
- Doses / Concentrations:
500 mg/kg
Basis:
nominal conc. - No. of animals per sex per dose:
- 3 males/ time period
- Control animals:
- yes
- Positive control(s):
- dimethylnitrosamine (DMN), dissolved in water
- Route of administration: orally
- Doses / concentrations: 10 mg/kg - Tissues and cell types examined:
- hepatocytes
- Details of tissue and slide preparation:
- see "any other information on materials and methods"
- Statistics:
- One-way analysis of variance was performed on UDS and S-phase data with multiple treatment groups. S-phase data were adjusted by square root transformation. Treatment means were compared to control means by Dunnett's multiple comparison test. Level of significance was <0.05.
- Key result
- Sex:
- male
- Genotoxicity:
- negative
- Remarks:
- TMP did not induce unscheduled DNA synthesis in hepatocyte cultures from rats treated in vivo.
- Toxicity:
- no effects
- Remarks:
- Mean cell viabilities in preparations of hepatocytes were: 84 % at 2-12 hours after administration of 500 mg/kg TMP.
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- see "Remarks on results"
- Conclusions:
- Interpretation of results: negative
Based on this study design 2, 2, 4- trimethyl pentane does not induce unscheduled DNA synthesis in vivo by unchanged application via gavage. - Executive summary:
Based on this study design 2, 2, 4- trimethyl pentane does not induce unscheduled DNA synthesis in vivo by unchanged application via gavage.
- Endpoint:
- in vivo mammalian germ cell study: cytogenicity / chromosome aberration
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- July 1978 - January 1980
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline with acceptable restrictions.
- Justification for type of information:
- A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 478 (Genetic Toxicology: Rodent Dominant Lethal Test)
- Principles of method if other than guideline:
- Similar to OECD 478, only two dose levels were used.
- GLP compliance:
- not specified
- Type of assay:
- rodent dominant lethal assay
- Species:
- rat
- Strain:
- Sprague-Dawley
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Breeding Laboratories, Wilmington, Massachusetts
- Age at study initiation: males: 10 weeks at initiation of pre-treatment mating period; females: 9 weeks at initiation of week 1 of pre-treatment mating period; females: 8 weeks at initiation of week 1 of post-treatment mating period
- Housing: males: individually housing during the treatment period; females: individually housing during the pre- and post-mating period
- Diet: Purina Laboratory Chow; ad libitum; except chamber-exposed groups during the treatment period
- Water: ad libitum; except chamber-exposed groups during the treatment period
- Route of administration:
- inhalation
- Vehicle:
- none
- Details on exposure:
- TYPE OF INHALATION EXPOSURE: whole body
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The stainless steel and glass chambers had a total volume of one cubic metre with an effective volume of 760 L.
- Air flow rate: 132 L/min
- Air change rate: every 7.6 min
- The test substance was placed in fritted-bottom, gas-washing bottles. Dry air, at various flow rates, was passed through the test material and the vapour-air mixtures entered the chamber air inlets and were further diluted with room air to provide the desired concentrations.
TEST ATMOSPHERE
Atmospheric sampling was performed using a Wilks Scientific Corp., Miran IA Ambient Air Analyzer (long pathlength infrared). On each exposure day three samples were drawn from each exposure chamber (at 1, 3 and 5 hours) and the exposure concentrations calculated by comparing the absorption of this sample to the standard curve. - Duration of treatment / exposure:
- 8 consecutive weeks
- Frequency of treatment:
- 6 hours/day, 5 days/week
- Post exposure period:
- 4 weeks
- Remarks:
- Doses / Concentrations:
400 and 1200 ppm
Basis:
nominal conc. - No. of animals per sex per dose:
- only males were exposed: 10/group
- Control animals:
- yes, sham-exposed
- Positive control(s):
- triethylenemelamine
- Route of administration: intraperitoneally on a single day, at least 2 hours prior to mating
- Doses level/ volume: 0.5 mg/kg body weight; 1 mL/kg body weight
- Vehicle: 0.9 % injectable normal saline solution - Tissues and cell types examined:
- Males: testes, seminal vesicles, epididymides, prostate tissues
Females: uteri - Details of tissue and slide preparation:
- METHOD OF ANALYSIS:
Males: Seminal vesicles, epididymides, prostate and any abnormal lesion or tissue masses were preserved in 10 % neutral buffered formalin. Testes were stored for 48 hours in Bouin´s solution and subsequently placed in formalin. Tissues from 5 randomly chosen males from each group were examined.
Females: The uteri from all females (pregnant and non-pregnant) were appropriately identified and preserved intact in a 10 % neutral buffered formalin solution. - Statistics:
- Comparisons were made during the treatment and post-treatment periods between negative control, positive control and test substance-treated groups by the Chi-square test. Absolute data were compared by using the F-test and Student´s t-test. When variances differed significantly, Student´s t-test was appropriately modified using Cochran´s approximation (t´). Corpora lutea and implantation data were evaluated by the one-tailed test.
- Key result
- Sex:
- male/female
- Genotoxicity:
- negative
- Remarks:
- At dose level of 400 and 1200 ppm the test substance was not mutagenic in rats by the dominant-lethal test.
- Toxicity:
- no effects
- Vehicle controls validity:
- other: no vehicle was used
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results: negative
Under the conditions of the test no negative effects were induced by the test substance, hydrocarbons, C7-C9, isoalkanes. - Executive summary:
Under the conditions of the test no negative effects were induced by the test substance, hydrocarbons, C7 -C9, isoalkanes.
Referenceopen allclose all
Mean cell viabilities in preparations of hepatocytes were: 85 % at 24 hours after administration of water (negative control) and 84 % at 2-12 hours after administration of 500 mg/kg TMP.
No increase in UDS expressed as increase in NG or number of cells in repair was observed in hepatocytes isolated from male rats at 2, 12, 24, or 48 hours post-dose. A strong response was elicited by the DMN positive control, as expected.
Autoradiographic preparations of hepatocytes isolated from rats 24 hr after treatment with TMP revealed a significant increase in the number of cells in S phase (2.1% vs 0.35% in controls); this value dropped back to 0.55% at 48 hours. TMP also significantly increased RDS in mice at 24 hours; no 48 hour values were presented. However low serum alanine transaminase activity were observed at 24 and 48 hours after a single treatment with 500 mg/kg TMP indicating that TMP induced little if any hepatocellular necrosis which could trigger regenerative cell proliferation.
Following 11 consecutive daily treatments with 100 mg/kg/day TMP, a significant increase (p<0.05) in rat hepatic concentration of DNA (mg/g liver) was seen but the total hepatic DNA content per rat was comparable to controls. Liver weight relative to body weight was also comparable to controls. The demonstrated increase in RDS suggests that TMP may stimulate additive rather than regenerative cell proliferation but not of sufficient magnitude to significantly increase the total DNA content of the liver.
TMP was administered at a single 500 mg/kg concentration to evaluate UDS and RDS in conjunction with a multidose study for unleaded gasoline. When RDS was observed, a liver DNA content determination was performed in rats with treatment over 11 days to determine the type of proliferative response.
No treatment-related effects were observed for mortality, in-life physical or necropsy examinations and for body weights in males.
No treatment effect was indicated from microscopic evaluation of testes, seminal vesicles, epididymides and prostate tissues from 5 randomly chosen males from each exposure group.
In one animal of the 400 ppm exposure group a caseous mass was observed in the left epididymis. This caseous mass was seen microscopically to be a spermatic granuloma. Since this was observed in a single animal, this is not considered to be treatment related. Several animals (one in control, one in 100 ppm group, one in 1200 ppm group) presented a purulent prostatitis which was of probable bacterial etiology.
Pregnancy rates, implantation data and implantation efficiency values and fetal death data for females mated to treated males were considered comparable to data for females mated to negative control males for each week of the post-treatment mating period. At dose level of 400 and 1200 ppm the test substance was not mutagenic in rats by the dominant-lethal test.
Positive control:
Females mated to the positive control males had fewer implants and lower implantation efficiency values (indicative of pre-implantation loss) than females mated to the negative control males. Fetal death parameters (mean and expressed as a percentage of total implants) for these same females were significantly increased during the post-treatment mating interval.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
There is no in vitro or in vivo genetic toxicity data available for Isoheptane. However, data is available for structural analogues, Heptane; Hydrocarbons, C7-C9, isoalkanes; and 2,2,4-trimethylpentane and presented in the dossier. This data is read across to Isoheptane based on analogue read across and a discussion and report on the read across strategy is provided as an attachment in IUCLID Section 13.
In vitro
In Vitro Gene Mutation in Bacteria
Heptane
The in vitro genotoxicity of n-heptane (CAS No. 142-82-5) has been assessed in different test systems. A bacterial reverse mutation assay (Ames test) was conducted with n-heptane following a protocol similar to OECD 471. The pre-incubation procedure was performed with Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100, and Escherichia coli strains WP2 and WP2 uvr A. The strains were exposed to concentrations of 0, 3.91, 7.81, 15.6, 31.3, 62.5, 125, and 250 µg/mL for 48 -72 h both with and without metabolic activation. No significant increases in the ratio of mutations over controls was seen. Therefore, the test substance was not mutagenic in either the presence or absence of metabolic activation (Shell Chemicals, 1983; Brooks et al., 1988).
2,2,4-trimethylpentane
A bacterial reverse mutation assay (Ames test) was conducted with iso-octane following a protocol similar to OECD 471. Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538 were exposed to the test substance at 20, 60, 560, 1670, and 5000 µg/plate in the absence and in the presence of a metabolic activator. Cytotoxic effects were observed at 5000 µg/plate. Iso-octane did not induce mutations in the bacterial mutation test in either the absence or presence of metabolic activator in any strain tested. Under the test conditions, the experimental compound, iso-octane, did not exhibit a positive response and is, therefore, considered not be mutagenic in this system (Chevron Phillips, 1982).
In Vitro Chromosome Aberration in Mammalian Cells
Heptane
The potential of n-heptane to cause chromosomal aberrations in rat liver RL4 cells was tested with a method comparable to OECD 473. Cells were exposed to concentrations of 0, 2.5, 5, and 10 µg/mL of test substance for 22 h, and then examined for chromosomal aberrations including polyploidy, chromatid gaps, and chromatid exchanges. No cytotoxicity was observed. A significant increase in chromatid gaps was seen at 10 µg/mL. However, this effect was neither dose-dependent nor accompanied by an increase in any other aberration type, and therefore not considered to be treatment-related. Under the conditions of this study, the test material was not clastogenic (Shell Chemicals, 1983; Brooks et al., 1988).
In Vitro Gene Mutation in Mammalian Cells
2,2,4-trimethylpentane
Isooctane (CAS No. 540-84-1) was tested in a mammalian cell gene mutation assay performed according to OECD 476. The test material was prepared by adding iso-octane at a final concentration of 5 % v/v in culture (DMEM) medium and stirred overnight at room temperature in a foil wrapped, capped parafilm-sealed bottle to saturate the medium. Human lymphoblastoid cells (TK6) were exposed to 100 or 50% of this saturated DMEM medium with and without metabolic activation for 3 h and allowed for expression for 4 to 8 days. Both with and without metabolic activation, iso-octane did not induce significant increases in the mutation frequency at the thymidine kinase locus and cell survival in iso-octane-saturated medium was greater than 50-60%. Based on the study design there was no incidence of increased genetic toxicity caused by the test substance (Richardson et al., 1986).
Isooctane was also tested in a Mouse Lymphoma Forward Mutation Assay performed according to a protocol similar to OECD TG 476. At concentrations ranging from 20 to 500 µg/mL (limit of solubility), iso-octane did not induce gene mutations in the mouse lymphoma L5178Y heterozygous TK+/-cells in the presence of metabolic activation, resulting in cytotoxicity at 165 µg/mL and above. Without metabolic activation, cytotoxicity at 500 µg/mL and an equivocal to weakly mutagenic response were observed (Chevron Phillips, 1982).
In Vitro Sister Chromatid Exchange in Mammalian Cells
2,2,4-trimethylpentane
Human lymphoblastoid cells exposed to iso-octane for 3 h as described above were also tested for the induction of sister chromatid exchange (SCE) according to OECD TG 479. Following exposure, the cells were incubated in the dark in the presence of 5'Bromo-deoxyuridine (10 µM) for 36 h, and metaphases were analysed microscopically thereafter. Both with and without metabolic activation, iso-octane did not induce an increased incidence of SCEs in TK6 cells (Richardson et al., 1986; Wilmer et al., 1981).
No statistically significant increase in number of SCEs (with and without metabolic activation) was also observed in another study compliant with OECD TG 479. Chinese hamster Ovary (CHO) cells were exposed to 25, 84, 250 µg/mL iso-octane in DMSO and 1400 and 3500 µg/mL iso-octane without vehicle. No cell growth occurred at the 250 µg/mL, without activation. Based on the study design there was no increased mutagenic response caused by isooctane in CHO cells with and without metabolic activation (Chevron Phillips, 1982).
In Vitro Unscheduled DNA Synthesis in Mammalian Cells
2,2,4-trimethylpentane
The potential of isooctane to induce unscheduled DNA synthesis (UDS) as an indicator of genotoxicity was tested in rat hepatocytes in a study comparable to OECD TG 482. Primary cultures were prepared from hepatocytes isolated from a rat. After attachment to coverslips and acclimatisation, cells were treated with [3H] thymidine (10 µCi/mL) and undiluted iso-octane at 0.33, 1.00, 3.33 % (v/v). The cells were then incubated for 18 hours. The highest concentration of 3.33% v/v was lethal to cell cultures. At the lower concentrations, isooctane did not induce unscheduled DNA synthesis in vitro by direct application to primary hepatocyte cultures. Based on the study design there was no incidence of increased genetic toxicity caused by the test substance (Loury et al. 1986).
In Vivo
Hydrocarbons, C7-C9, isoalkanes
Hydrocarbons, C7-C9, isoalkanes tested in a dominant lethal study (similar to OECD 478) showed no evidence of genotoxicity in the germ cells of treated male rats exposed to 400 or 1200 ppm by inhalation (ExxonMobil Chemical,1980).
2,2,4-trimethylpentane
As part of an unscheduled DNA synthesis (UDS) study performedin vitro, iso-octane was administered to rats and mice orally at a single dose of 500 mg/Kg. Primary hepatocytes were isolated from livers at sacrifice times 2, 12, 28 or 48 hours post-treatment and cultures were evaluated for unscheduled and replicative DNA synthesis. Iso-octane did not induce unscheduled DNA synthesis in hepatocyte cultures, and increases in replicative DNA synthesis were not of sufficient magnitude to increase the total DNA content of the liver (Loury et al., 1986).
Justification for classification or non-classification
The negative results in read across in vitro and in vivo genotoxicity assays do not warrant the classification of Isoheptane as genotoxic under the new Regulation (EC) 1272/2008 on classification, labeling and packaging of substances and mixtures (CLP).
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