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EC number: 930-397-4 | CAS number: 1174918-63-8
- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
There is no in vitro genetic toxicity data available for Hydrocarbons, C5-C7, n-alkanes, isoalkanes, n-hexane rich. However, data is available for structural analogue 5-80% n-hexane and presented in the dossier. This data is read across to Hydrocarbons, C5 -C7, n-alkanes, isoalkanes, n-hexane rich based on analogue read across and a discussion and report on the read across strategy is provided as an attachment in IUCLID Section 13.
The read across genetic toxicity tests listed below had negative results for Hydrocarbons, C5 -C7, n-alkanes, isoalkanes, n-hexane rich.
Genetic Toxicity in vitro – Bacterial reverse mutation assay (OECD 471)
Genetic Toxicity in vitro – Mammalian Chromosome Aberration Test (OECD TG 473)
Genetic Toxicity in vitro – Mammalian Cell Gene Mutation Test (OECD TG 476)
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:
- 1989-05-19 to 1989-07-06
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: This study is classified as reliable without restriction because it followed a protocol comparable to OECD Guideline 471 but was modified to test the vapor phase of the test substance.
- 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:
- The method was modified to use the dessicator methodology in order to test the mutagenicity of the vapor phase of the test substance.
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98, TA 100
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 from aroclor induced rat liver
- Test concentrations with justification for top dose:
- 0, 600, 1000, 3000, 6000, 9000 ppm
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- other: no solvent used
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene, 2-nitrofluorene, sodium azide, 9-aminoacridine, 1,1-dichloroethene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: Prepared plates were placed uncovered and inverted in 9l dessicators. An appropriate amount of test substance was placed in a glass petri dish was suspended at the bottom of the dessicator. A magnetic stirring bar was placed at the bottom of the dessicator, and served as a fan to ensure even distribution of the vapor.
DURATION
- Preincubation period: 48 hrs
- Exposure duration: 7-8 hrs at 37 degree C
- Expression time (cells in growth medium): There was an additional incubation time of 40 hrs after exposure.
NUMBER OF REPLICATIONS: 3
- Evaluation criteria:
- To be considered positive for mutation, at least a doubling of mean revertants per plate in at least one tester strain must be seen. This increase must be dose-related.
- Statistics:
- Mean number of revertant per plate and the standard deviation was calculated.
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not examined
- 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
- Vehicle controls validity:
- not examined
- Untreated negative 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
- Vehicle controls validity:
- not examined
- 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
- Vehicle controls validity:
- not examined
- 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
- Vehicle controls validity:
- not examined
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- No positive responses were observed in any of the tester strains. The positive control substance, 2-aminoanthracene, did not produce any mutations in strain TA 1538 in one experiment in the presence of S9. As revertants were seen in the test of TA 1538 without S9, and revertants were seen in other strains exposed to positive controls with S9, the lack of revertants was most likely due to a technical error and the study is still considered valid.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
The test substance is not mutagenic. - Executive summary:
This study examined the mutagenicity of vapors of the test substance commercial hexane. Plates of S. typhimurium were exposed for 7 -8 hrs to test atmospheres of 0, 600, 1000, 3000, 6000, or 9000 ppm of test substance. 20,000 ppm of 1,1 -dichloroethene was used a vapor-phase positive control substance. The test substance did not produce a positive response in any of the test strains. The test substance is not mutagenic.
- 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:
- 1989-08-24 to 1990-02-05
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: This study is classified as reliable without restriction because it followed a protocol comparable to OECD Guideline 473.
- 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
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Additional strain / cell type characteristics:
- other: CHO-K1 cells
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 from aroclor induced rat liver
- Test concentrations with justification for top dose:
- 0.015, 0.034, 0.074, 0.123, 0.416 ul/ml without S9
0.014, 0.022, 0.056, 0.118, 0.251 ul/ml with S9 - Vehicle / solvent:
- DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: triethylenemelamine 0.5 ug/ml without S9, cyclophosphamide 50 ug/ml with S9
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: 100 ul of dosing solutiong was added to the test medium
DURATION
- Preincubation period: 16-24 hrs at 37 degree C
- Exposure duration: 12 hrs without S9 at 37 degree in humidified air; 2 hrs with S9 at 37 degree in humidified air
- Expression time (cells in growth medium): For cells not treated with S9, two hours prior to cell harvest, treatment medium was removed and cells washed with PBS and refed with medium containing 0.1 ug/ml of Colcemid. For cells treated with S9, treatment medium was removed after exposure, cells were washed with PBS, refed, and returned to the incubator for 16 hrs. Colcemid was added at 0.1 ug/ml, and flasks incubated for two hrs.
- Fixation time (start of exposure up to fixation or harvest of cells): 14-20 hrs, collected by centrifugation
SELECTION AGENT (mutation assays):
SPINDLE INHIBITOR (cytogenetic assays):
STAIN (for cytogenetic assays): 5% Giemsa
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 100 per concentration
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index and cell cycle delay
OTHER EXAMINATIONS:
Chromatid and isochromatid breaks, exchange figures, chromosome breaks, fragments, pulverized chromosomes, chromatid and isochromatid gaps
OTHER: - Evaluation criteria:
- In order for the test to be valid, there must be no more than 6% cells with chromosome aberrations in the negative and solvent control groups. Positive control must be statistically increased over untreated controls (p<=0.05, Fisher's exact test). A positive result is percentage of cells with aberrations was statistically increased over untreated controls (p<=0.05, Fisher's exact test).
- Statistics:
- Fisher's exact test was used to determine statistical significance. Cochran-Armitage test was used to test dose-responsiveness.
- Key result
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- >= 0.074 ul/ml
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Toxicity was observed at concentrations of 0.074 ul/ml or greater. No significant increase in chromosome aberrations was seen in treatment groups.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
The test substance is not clastogenic. - Executive summary:
This study examined the potential for commercial hexane to cause chromosome aberrations in Chinese Hamster Ovary (CHO) cells. CHO cells were exposed to concentrations of 0, 0.015, 0.034, 0.074, 0.123, and 0.416 ul/ml without metabolic activation and 0, 0.014, 0.022, 0.056, 0.118, and 0.251 ul/ml with metabolic activation. 0.5 ug/ml triethylenemelamine was used a positive control without metabolic activation and 50 ug/ml cyclophosphamide was used as a positive control with metabolic activation. Negative and positive controls were valid. There was no significant increase in chromosome aberrations in any test group. The test substance was cytotoxic at concentrations of 0.074 ul/ml or greater. The test substance is 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
- Study period:
- Aug. 28, 1989-April 2, 1990
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: This study is classified as reliable without restriction because it closely followed a protocol comparable to OECD Guideline 476.
- 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 476 (In Vitro Mammalian Cell Gene Mutation Test)
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- mammalian cell gene mutation assay
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- Periodically checked for Mycoplasma contamination: yes- Additional strain / cell type characteristics:
- other: K1-BH4
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 from aroclor induced rat liver
- Test concentrations with justification for top dose:
- 0, 0.132, 0.098, 0.063, 0.0362, 0.0122 ul/ml (analytical)
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: ethylmethanesulphonate 0.2 ul/ml without metabolic activation, benzo(a)pyrene 4 ug/ml with metabolic activation
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: 7.5 ml of test substance was added to 17.5 ml of DMSO and vortexed for 2 min. The solution was allowed to separate, and the bottom DMSO/test substance layer and diluted further with DMSO. Appropriate amounts were then added to the test medium.
DURATION
- Preincubation period: 18-24 hrs at 37 degree C
- Exposure duration: 5 hrs at 37 degree C
- Expression time (cells in growth medium): 18-24 at 37 degree C
NUMBER OF REPLICATIONS: 2
DETERMINATION OF CYTOTOXICITY
Replicates from each treatment concentration were pooled and cultured in triplicate (100 cells/60 mm dish). After 7 days of incubation, colonies were fixed with methanol , stained with Giemsa, and counted. Determination was based on relative cloning efficiency. - Evaluation criteria:
- mutant frequency > 20 mutants per 10^6 clonable cells, at least twice the mutant frequency of negative and solvent controls, mutant frequency above negative and solvent controls of at least 11 mutants per 10^6 clonable cells
- Statistics:
- Students t-test (p <= 0.05)
- Key result
- Species / strain:
- Chinese hamster Ovary (CHO)
- 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:
- None of the mutant frequencies in the test groups were significantly elevated over negative and solvent controls. The test substance was cytotoxic at concentrations of 0.063 ul/ml or greater.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
The test substance is not mutagenic. - Executive summary:
This study determined the mutagenicity of commercial hexane to Chinese hamster ovary (CHO) cells. CHO cells were exposed to concentrations of 0, 0.132, 0.098, 0.063, 0.0362, or 0.0122 ul/ml both with and without metabolic activation for 5 hrs. The cells were then analyzed for mutation frequency. A concurrent cytotoxicity assay was performed. Results of the mutagenicity assay show that the test substance is not mutagenic both in the presence and absence of metabolic activation. However, the test substance was cytotoxic at concentrations of 0.063 ul/ml or greater.
Referenceopen allclose all
Average Revertants per Plate (SD) - Experiment B1
Dose (ppm) |
TA 98 |
TA 100 |
TA 1535 |
TA 1537 |
TA 1538 |
Without S9 |
|||||
0.0 |
17 ± 1 |
105 ± 12 |
11 ± 1 |
7 ± 1 |
8 |
600 |
20 ± 5 |
113 ± 9 |
13 ± 4 |
6 ± 1 |
6 |
1000 |
16 ± 3 |
121 ± 8 |
14 ± 2 |
7 ± 1 |
6 |
3000 |
18 ± 4 |
112 ± 13 |
17 ± 3 |
5 ± 4 |
4 |
6000 |
24 ± 4 |
117 ± 14 |
12 ± 4 |
6 ± 4 |
8 |
9000 |
17 ± 3 |
112 ± 7 |
14 ± 5 |
9 ± 2 |
5 |
Positive control |
227 ± 8 |
422 ± 44 |
349 ± 36 |
481 ± 135 |
452 |
With S9 |
|||||
0.0 |
24 ± 5 |
119 ± 10 |
17 ± 1 |
7 ± 0 |
12 |
600 |
30 ± 10 |
137 ± 12 |
23 ± 4 |
9 ± 4 |
15 |
1000 |
23 ± 5 |
126 ± 3 |
14 ± 2 |
11 ± 5 |
13 |
3000 |
22 ± 1 |
120 ± 19 |
16 ± 2 |
9 ± 2 |
9 |
6000 |
24 ± 2 |
103 ± 11 |
14 ± 4 |
7 ± 2 |
11 |
9000 |
30 ± 6 |
120 ± 3 |
17 ± 1 |
8 ± 2 |
14 |
Positive control |
3142 ± 139 |
3902 ± 68 |
189± 22 |
351 ± 26 |
-- |
Positive vapor control |
372 ± 52 |
Average Revertants per Plate (SD) - Experiment B2
Dose (ppm) |
TA 98 |
TA 100 |
TA 1535 |
TA 1537 |
TA 1538 |
Without S9 |
|||||
0.0 |
12 ± 2 |
107 ± 11 |
9 ± 5 |
5 ± 1 |
19 |
600 |
20 ± 1 |
103 ± 9 |
8 ± 4 |
7 ± 3 |
19 |
1000 |
14 ± 4 |
110 ± 6 |
12 ± 5 |
7 ± 2 |
19 |
3000 |
14 ± 1 |
124 ± 14 |
12 ± 2 |
7 ± 2 |
19 |
6000 |
22 ± 6 |
125 ± 14 |
15 ± 1 |
5 ± 2 |
16 |
9000 |
15 ± 6 |
114 ± 5 |
13 ± 3 |
5 ± 3 |
13 |
Positive control |
444 ± 86 |
991 ± 67 |
758 ± 19 |
162 ± 51 |
754 |
With S9 |
|||||
0.0 |
21 ± 4 |
132 ± 16 |
17 ± 4 |
7 ± 3 |
26 |
600 |
25 ± 7 |
133 ± 8 |
18 ± 2 |
6 ± 1 |
25 |
1000 |
22 ± 2 |
155 ± 4 |
15 ± 7 |
8 ± 3 |
25 |
3000 |
20 ± 1 |
123 ± 23 |
16 ± 7 |
7 ± 3 |
24 |
6000 |
21 ± 4 |
151 ± 14 |
15 ± 1 |
6 ± 2 |
30 |
9000 |
25 ± 4 |
161 ± 8 |
11 ± 4 |
6 ± 1 |
27 |
Positive control |
2187 ± 166 |
2229 ± 223 |
166 ± 32 |
230 ± 24 |
2195 |
Positive vapor control |
394 ± 50 |
Results of CHO Chromosome Aberration Assay
Dose (µl/ml) |
Mitotic Index |
Cells Scored |
Aberrations per cell (mean ± SD) |
% Cells with Aberrations |
Without S9 |
||||
Negative Control |
5.7 |
100 |
0.010 ± 0.100 |
1 |
Solvent Control |
4.9 |
100 |
0.010 ± 0.100 |
1 |
0.015 |
4.8 |
100 |
0.000 ± 0.000 |
0 |
0.034 |
4.5 |
100 |
0.010 ± 0.100 |
1 |
0.074 |
2.9 |
100 |
0.010 ± 0.100 |
1 |
0.123 |
0.2 |
8 |
0.000 ± 0.000 |
0 |
0.416 |
0.0 |
0 |
||
Positive Control |
2.2 |
100 |
0.360 ± 1.124 |
21 |
With S9 |
||||
Negative Control |
5.3 |
100 |
0.010 ± 0.100 |
1 |
Solvent Control |
5.6 |
100 |
0.010 ± 0.100 |
1 |
0.014 |
5.3 |
100 |
0.010 ± 0.100 |
1 |
0.022 |
5.9 |
100 |
0.010 ± 0.100 |
1 |
0.056 |
3.1 |
100 |
0.010 ± 0.100 |
1 |
0.118 |
0.2 |
2 |
0.000 ± 0.000 |
0 |
0.251 |
0.0 |
0 |
||
Positive Control |
2.5 |
100 |
0.840 ± 1.819 |
40 |
CHO/HGPRT Mutation Assay
Dose (µl/ml) |
Total Colonies |
Cloning Efficiency |
Total Mutant Colonies |
Mutants/106 Clonable Cells |
Without S9 |
||||
Negative control |
310 |
1.03 |
19 |
18.4 |
Solvent Control |
329 |
1.10 |
6 |
5.5 |
0.132 |
284 |
0.95 |
0 |
5.3 |
0.098 |
260 |
0.87 |
0 |
5.8 |
0.063 |
321 |
1.07 |
0 |
0.9 |
0.0362 |
289 |
0.96 |
5 |
5.2 |
0.0122 |
294 |
0.98 |
12 |
12.2 |
Positive Control |
282 |
0.94 |
227 |
241.5 |
With S9 |
||||
Negative control |
320 |
1.07 |
0 |
0.9 |
Solvent Control |
330 |
1.10 |
1 |
0.9 |
0.132 |
299 |
1.00 |
0 |
1.0 |
0.098 |
299 |
0.99 |
0 |
1.0 |
0.063 |
256 |
0.85 |
4 |
4.7 |
0.0362 |
317 |
1.06 |
13 |
12.3 |
0.0122 |
293 |
0.98 |
13 |
13.3 |
Positive Control |
239 |
0.80 |
91 |
114.2 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
There is no in vivo genetic toxicity data available for Hydrocarbons, C5 -C7, n-alkanes, isoalkanes, n-hexane rich. However, data is available for structural analogue 5-80% n-hexane and presented in the dossier. This data is read across to Hydrocarbons, C5-C7, n-alkanes, isoalkanes, n-hexane rich based on analogue read across and a discussion and report on the read across strategy is provided as an attachment in IUCLID Section 13.
The read across genetic toxicity tests listed below had negative results for Hydrocarbons, C5 -C7, n-alkanes, isoalkanes, n-hexane rich.
Genetic Toxicity in vivo – Mammalian Bone Marrow Chromosome Aberration Test (equivalent/similar to OECD 475)
Genetic Toxicity in vivo – Rodent Dominant Lethal Test (equivalent/similar to OECD 478)
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / bone marrow 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:
- 1990
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: This study is classified as reliable without restriction because it closely followed OECD Guideline 475.
- 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
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- In order to test substance as a vapor, animals were exposed via nose-only inhalation.
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- other: cytogenetics assay
- Species:
- rat
- Strain:
- Sprague-Dawley
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Harlan Sprague Dawley, Inc., Frederick, MD
- Age at study initiation: 6-8 weeks
- Weight at study initiation: 228-266 g males, 153-188 g females
- Assigned to test groups randomly: assigned using body weight randomization program
- Housing: singly in plastic cages, identified by ear tags
- Diet (e.g. ad libitum): certified laboratory rodent chow, ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: 10 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 72 degree F
- Humidity (%): 40-70
- Photoperiod (hrs dark / hrs light): 12 hrs light/12 hrs dark
- Route of administration:
- inhalation: vapour
- Details on exposure:
- TYPE OF INHALATION EXPOSURE: nose only
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 360 ml polycarbonate chamber
- Method of holding animals in test chamber: Polycarbonate tubes were used to restrain the animals. Only the nose of the animals protruded into the chambers. A soft sponge was used as a plunger to hold the animals in place.
- Source and rate of air: filtered outside air
- Method of conditioning air: A Gilson peristaltic pump was used to meter test liquid in sealed resevoirs to vaporization flasks immersed in 57 degree C water. Air was passed through the flasks, then through teflon tubing to the exposure chamber. A calibrated rotameter was used to dilute the air to the appropriate concentration.
- Temperature, humidity, pressure in air chamber: monitored every 30 min.
- Air flow rate: monitored every 30 min.
TEST ATMOSPHERE
- Brief description of analytical method used: GC
- Samples taken from breathing zone: yes - Duration of treatment / exposure:
- 6 hrs per day
- Frequency of treatment:
- 5 days
- Post exposure period:
- 1 or 19 hrs after end of exposure
- Remarks:
- Doses / Concentrations:
0, 900, 3000, 9000 ppm (0, 3168, 10560, 31680 mg/m3)
Basis:
nominal conc. - No. of animals per sex per dose:
- 5 male and 5 female per dose
- Control animals:
- yes, sham-exposed
- Positive control(s):
- triethylenemelamine
- Route of administration: intraperitoneal injection
- Doses / concentrations: 0.5 mg/kg - Tissues and cell types examined:
- bone marrow
- Details of tissue and slide preparation:
- TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):
Colchicine was administered two hours prior to sacrifice to arrest cell division at metaphase. Animals were sacrificed by carbon dioxide asphyxiation. Bone marrow cells were removed from the femur by aspiration into HBSS. Cells were mixed well and kept in an ice bath until all samples were collected. After centrifuging for 10 min, the supernatent was discarded, and the cells resuspended in 5 ml 0.075 M KCl at 37 degree C. After incubating for 10 min, cells were again centrifuged and resuspended in fixative. This was repeated with fresh fixative.
DETAILS OF SLIDE PREPARATION:
Cells were again centrifuged for 10 min, the supernantant decanted, and the cells resuspended in 1 ml of fixative. Two drops were placed on a cold wet glass slide, and allowed to air dry. Three slides were prepared per animal. Slides were stained with 4% Giemsa.
METHOD OF ANALYSIS:
50 metaphase cells containing approx. 40 centromeres were scored if possible.
OTHER: - Evaluation criteria:
- In order for the test to be valid, there must be no more than 4% cells demonstrating aberrations in the negative control group. Positive control must be statistically increased over untreated controls (p<=0.05, Fisher's exact test). A positive result is a percentage of cells with aberrations which was statistically increased over untreated controls (p<=0.05, Fisher's exact test).
- Statistics:
- Fisher's exact test was used to determine statistical significance. Cochran-Armitage test was used to test dose-responsiveness.
- Key result
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- There was a decrease in body weight observed after the last exposure. This reduction is typical for nose-only exposure, and is considered to be due to restraint. There was no statistically significant increase in aberrations in the exposure groups.
- Conclusions:
- Interpretation of results: negative
The test substance is not mutagenic. - Executive summary:
This study determined the effect of inhalation exposure of commercial hexane on rat bone marrow. Groups of 5 male and 5 female rats were exposed to 0, 900, 3000, and 9000 ppm of test substance vapor for 6 hrs/day for 5 days. 0.5 mg/kg triethylenemelamine was used as a positive control substance. Animals were sacrificed 3 or 21 hrs after exposure, and the bone marrow from their femurs examined for cell aberrations. There was no statistically significant increase in cell aberrations in any treatment group. The test substance is not mutagenic.
Reference
Chromosomal Damage in Rat Bone Marrow - Male
Dose (ppm) |
Time Evaluated (hrs) |
Cells with Aberrations (%) |
Mean Aberrations per Cell per Animal |
0 |
6 |
0.0 |
0.000 ± 0.000 |
0 |
24 |
0.0 |
0.000 ± 0.000 |
876 |
6 |
0.4 |
0.004 ± 0.009 |
876 |
24 |
0.0 |
0.000 ± 0.000 |
3249 |
6 |
0.0 |
0.000 ± 0.000 |
3249 |
24 |
0.4 |
0.004 ± 0.009 |
8715 |
6 |
0.8 |
0.008 ± 0.018 |
8715 |
24 |
0.4 |
0.004 ± 0.009 |
Positive Control |
6 |
14.4 |
0.696 ± 0.121 |
Chromosomal Damage in Rat Bone Marrow - Female
Dose (ppm) |
Time Evaluated (hrs) |
Cells with Aberrations (%) |
Mean Aberrations per Cell per Animal |
0 |
6 |
0.8 |
0.008 ± 0.011 |
0 |
24 |
0.0 |
0.000 ± 0.000 |
876 |
6 |
0.4 |
0.004 ± 0.009 |
876 |
24 |
0.4 |
0.004 ± 0.009 |
3249 |
6 |
0.4 |
0.004 ± 0.009 |
3249 |
24 |
0.0 |
0.000 ± 0.000 |
8715 |
6 |
0.4 |
0.004 ± 0.009 |
8715 |
24 |
0.4 |
0.004 ± 0.009 |
Positive Control |
6 |
20.8 |
0.900 ± 0.477 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
There is no genetic toxicity data available for Hydrocarbons, C5 -C7, n-alkanes, isoalkanes, n-hexane rich. However, data is available for structural analogue 5 -80% n-hexane and presented in the dossier. This data is read across to Hydrocarbons, C5-C7, n-alkanes, isoalkanes, n-hexane rich 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 study in bacteria
5 -80% n-hexane
In a key study (API, 1990a) the mutagenicity of vapors of the test substance commercial hexane was examined. Plates of S. typhimuriumwere exposed for 7 -8 hrs to test atmospheres of 0, 600, 1000, 3000, 6000, or 9000 ppm of test substance. 20,000 ppm of 1,1 -dichloroethene was used a vapor-phase positive control substance. The test substance did not produce a positive response in any of the test strains. The test substance is not mutagenic.
In vitro Chromosome Aberration in Mammalian Cells
5 -80% n-hexane
This study (API 1990c) examined the potential for commercial hexane to cause chromosome aberrations in Chinese Hamster Ovary (CHO) cells. CHO cells were exposed to concentrations of 0, 0.015, 0.034, 0.074, 0.123, and 0.416 ul/ml without metabolic activation and 0, 0.014, 0.022, 0.056, 0.118, and 0.251 ul/ml with metabolic activation. 0.5 ug/ml triethylenemelamine was used a positive control without metabolic activation and 50 ug/ml cyclophosphamide was used as a positive control with metabolic activation. Negative and positive controls were valid. There was no significant increase in chromosome aberrations in any test group. The test substance was cytotoxic at concentrations of 0.074 ul/ml or greater. The test substance is not clastogenic.
In vitro Gene Mutation study in Mammalian Cells
5 -80% n-hexane
This study (API, 1990b) determined the mutagenicity of commercial hexane to Chinese hamster ovary (CHO) cells. CHO cells were exposed to concentrations of 0, 0.132, 0.098, 0.063, 0.0362, or 0.0122 ul/ml both with and without metabolic activation for 5 hrs. The cells were then analyzed for mutation frequency. A concurrent cytotoxicity assay was performed. Results of the mutagenicity assay show that the test substance is not mutagenic both in the presence and absence of metabolic activation. However, the test substance was cytotoxic at concentrations of 0.063 ul/ml or greater.
In Vivo
In vivo Mammalian Bone Marrow Chromosome Aberration Test
5 -80% n-hexane
This study (Daughtrey, 1994) determined the effect of inhalation exposure of commercial hexane on rat bone marrow. Groups of 5 male and 5 female rats were exposed to 0, 900, 3000, and 9000 ppm of test substance vapor for 6 hrs/day for 5 days. 0.5 mg/kg triethylenemelamine was used as a positive control substance. Animals were sacrificed 3 or 21 hrs after exposure, and the bone marrow from their femurs examined for cell aberrations. There was no statistically significant increase in cell aberrations in any treatment group. The test substance is not mutagenic.
In vivo Rodent Dominant Lethal Test
Hexane
In a CD-1 mouse dominant lethal assay (Litton Bionectics, 1980), 12 male mice/group were treated via inhalation at doses of 0, 99.4 or 394.6 ppm n-hexane in filtered air (mean time-weighted average) for 6 hours per day, 5 days per week, for 8 weeks. Males were then mated for two weeks with untreated virgin females, and study authors compared fertility index, per-implantation loss, and post-implantation loss between treated and negative control groups. There were no signs of toxicity or mortality in male mice during the study.The positive control induced the appropriate response. The 394.6 -ppm group exhibited a small but significant increase in fertility index (proportion of pregnant females among mated females) during week two as compared to the negative control. The pre-implantation loss (number of implants per pregnant female in each group) was not significantly different from either dose group versus negative control. There were no significant differences in post-implantation loss (average number of dead implants per pregnant female) between the 99.4 -ppm dose group and negative control. There was a significant reduction in the average number of dead implants in the 394.6 ppm dose group as compared to negative control in week 1 and a slight increase during week two, but the result was not statistically significant. This result is not considered biologically significant because there is no corresponding increase in pre-implantation loss. Therefore, under conditions of the study, n-hexane is considered negative for inducing dominant lethal sperm cell mutations in male CD-1 mice.
Justification for classification or non-classification
The negative results of in vitro and in vivo genotoxicity assays from a structural analogue does not warrant the classification of Hydrocarbons, C5-C7, n-alkanes, isoalkanes, n-hexane rich as genotoxic under the new Regulation (EC) 1272/2008 on classification, labeling and packaging of substances and mixtures (CLP).
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
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